Patent Grant
9919072
The present invention relates to materials that may be used as wound dressings that may release nitric oxide. The present invention also relates to methods of making and using wound dressings that may release nitric oxide.
An important aspect for wound care is the control of infection, which may facilitate the healing process. Wound dressings are one of the most commonly used tools to protect the wound from infection. Antimicrobial agents are often incorporated into the wound dressing to treat and prevent infection. However, there are several disadvantages associated with use of antimicrobial agents. It has been observed that an increasing number of pathogens have developed resistance to the conventional antibiotic treatment. According to statistics, antibiotic-resistant pathogens are the primary reason for a majority of all lethal nosocomial infections. See Robson et al., Surg. Clin. N. Am. 77, 637-650 (1977). Furthermore, many antiseptic agents not only kill pathogens, but also impose a threat to the proliferating granulation tissue, fibroblasts and keratinocytes that may help with the wound healing process. Additionally, some antimicrobial agents may cause allergic reactions in some patients.
It is known that nitric oxide possesses a broad-spectrum of antimicrobial activity and may be used as an alternative to conventional antibiotics for drug resistant bacteria. Furthermore, some recent studies have demonstrated that nitric oxide may also play an important role in the wound healing process by promoting angiogenesis through stimulation of vascular endothelial growth factor (VEGF) and increased fibroblast collagen synthesis. See Schaffer M R, et al., Diabetes-impaired healing and reduced wound nitric oxide synthesis: A possible pathophysiologic correlation. Surgery 1997; 121(5):513-9; and Shi H P, et al., The role of iNOS in wound healing. Surgery 2001; 130 (2):225-9. Thus, nitric oxide presents a promising addition and/or alternative to the conventional antibiotic treatment for wound care.
Nitric oxide is a gas at ambient temperature and atmospheric pressure, and it has a short half-life in a physiological milieu. Several small molecule nitric oxide donor prodrugs have been developed which have contributed greatly to the understanding of nitric oxide in a number of disease states. However, due to issues with stability, indiscriminate NO-release, monotypical nitric oxide release kinetics, and inability to target specific tissue types no clinically viable solutions currently exist for administering nitric oxide outside of its gaseous form. Reproducibly delivering a the appropriate levels of nitric oxide for a given therapeutic indication is important because release of large amounts of nitric oxide may be toxic or create undesirable side effects such as decreases in angiogenesis or increased inflammation. Therefore, it has been challenging to use nitric oxide in the wound care field, other than via exogenous application, particularly in topical wound healing applications wherein nitric oxide has concentration dependent effects and benefits from delivery in a controlled and targeted manner.
Thus, the need exists for wound treatments and dressings that can release nitric oxide by a controlled delivery method.
Provided according to some embodiments of the invention are wound dressings that include a polymer matrix, and nitric oxide (NO)-releasing polysiloxane macromolecules within and/or on the polymer matrix. In some embodiments, such wound dressings are non-toxic and stably store NO. In some embodiments, the NO-releasing polysiloxane macromolecules include N-diazeniumdiolate functional groups and in some embodiments, include S-nitrosothiol functional groups.
In some embodiments of the invention, the concentration of the NO-releasing polysiloxane macromolecules is in a range of about 0.1 to about 20 weight percent.
The wound dressings may include additional additives. For example, the wound dressings may include a water-soluble porogen such as sodium chloride, sucrose, glucose, lactose, sorbitol, xylitol, polyethylene glycol, polyvinylpyrrollidone, polyvinyl alcohol and mixtures thereof. The wound dressings may also include at least one therapeutic agent such as antimicrobial compounds, anti-inflammatory agents, pain-relievers, immunosuppressants, vasodilators, wound healing agents, anti-biofilm agents and mixtures thereof.
In some embodiments, the wound dressing includes a polymer matrix that includes a hydrophilic polyurethane, such as, for example, an aliphatic polyether polyurethane that absorbs water in an amount ranging from 10 percent to 60 percent of its dry weight.
In some embodiments of the invention, the wound dressing includes a flexible, open-celled polyurethane foam that includes at least one polyisocyanate segment and at least one polyol segment. In some embodiments, the NO-releasing macromolecules are present within, and optionally crosslinked to, the polymer matrix of the polymer foam.
In some embodiments, the storage of nitric oxide in the wound dressing is in a range of 0.1 pmol NO cm−2 to 100 pmol NO cm−2, in some embodiments, in a range of 100 pmol NO cm−2 to 1000 pmol NO cm−2 and in some embodiments, in a range of 1 nmol NO cm−2 to 10 μmol NO cm−2.
In addition, provided according to some embodiments of the invention are wound dressing kits, methods of treating a wounds and methods of forming wound dressings.
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate certain embodiment(s) of the invention.
The foregoing and other aspects of the present invention will now be described in more detail with respect to the description and methodologies provided herein. It should be appreciated that the invention can be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used in the description of the embodiments of the invention and the appended claims, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. Also, as used herein, “and/or” refers to and encompasses any and all possible combinations of one or more of the associated listed items. Furthermore, the term “about,” as used herein when referring to a measurable value such as an amount of a compound, dose, time, temperature, and the like, is meant to encompass variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1% of the specified amount. It will be further understood that the terms “comprises” and/or “comprising,” when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. Unless otherwise defined, all terms, including technical and scientific terms used in the description, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
All patents, patent applications and publications referred to herein are incorporated by reference in their entirety. In the event of conflicting terminology, the present specification is controlling.
The embodiments described in one aspect of the present invention are not limited to the aspect described. The embodiments may also be applied to a different aspect of the invention as long as the embodiments do not prevent these aspects of the invention from operating for its intended purpose.
Chemical Definitions
As used herein the term “alkyl” refers to C1-20 inclusive, linear (i.e., “straight-chain”), branched, or cyclic, saturated or at least partially and in some cases fully unsaturated (i.e., alkenyl and alkynyl)hydrocarbon chains, including for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, octyl, ethenyl, propenyl, butenyl, pentenyl, hexenyl, octenyl, butadienyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, and allenyl groups. “Branched” refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl or propyl, is attached to a linear alkyl chain. Exemplary branched alkyl groups include, but are not limited to, isopropyl, isobutyl, tert-butyl. “Lower alkyl” refers to an alkyl group having 1 to about 8 carbon atoms (i.e., a C1-8 alkyl), e.g., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. “Higher alkyl” refers to an alkyl group having about 10 to about 20 carbon atoms, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. In certain embodiments, “alkyl” refers, in particular, to C1-5 straight-chain alkyls. In other embodiments, “alkyl” refers, in particular, to C1-5 branched-chain alkyls.
Alkyl groups can optionally be substituted (a “substituted alkyl”) with one or more alkyl group substituents, which can be the same or different. The term “alkyl group substituent” includes but is not limited to alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxyl, aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. There can be optionally inserted along the alkyl chain one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms, wherein the nitrogen substituent is hydrogen, lower alkyl (also referred to herein as “alkylaminoalkyl”), or aryl.
Thus, as used herein, the term “substituted alkyl” includes alkyl groups, as defined herein, in which one or more atoms or functional groups of the alkyl group are replaced with another atom or functional group, including for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, and mercapto.
The term “aryl” is used herein to refer to an aromatic substituent that can be a single aromatic ring, or multiple aromatic rings that are fused together, linked covalently, or linked to a common group, such as, but not limited to, a methylene or ethylene moiety. The common linking group also can be a carbonyl, as in benzophenone, or oxygen, as in diphenylether, or nitrogen, as in diphenylamine. The term “aryl” specifically encompasses heterocyclic aromatic compounds. The aromatic ring(s) can comprise phenyl, naphthyl, biphenyl, diphenylether, diphenylamine and benzophenone, among others. In particular embodiments, the term “aryl” means a cyclic aromatic comprising about 5 to about 10 carbon atoms, e.g., 5, 6, 7, 8, 9, or 10 carbon atoms, and including 5- and 6-membered hydrocarbon and heterocyclic aromatic rings.
The aryl group can be optionally substituted (a “substituted aryl”) with one or more aryl group substituents, which can be the same or different, wherein “aryl group substituent” includes alkyl, substituted alkyl, aryl, substituted aryl, aralkyl, hydroxyl, alkoxyl, aryloxyl, aralkyloxyl, carboxyl, acyl, halo, nitro, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acyloxyl, acylamino, aroylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylthio, alkylthio, alkylene, and —NR1R″, wherein R1 and R″ can each be independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, and aralkyl.
Thus, as used herein, the term “substituted aryl” includes aryl groups, as defined herein, in which one or more atoms or functional groups of the aryl group are replaced with another atom or functional group, including for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, and mercapto. Specific examples of aryl groups include, but are not limited to, cyclopentadienyl, phenyl, furan, thiophene, pyrrole, pyran, pyridine, imidazole, benzimidazole, isothiazole, isoxazole, pyrazole, pyrazine, triazine, pyrimidine, quinoline, isoquinoline, indole, carbazole, and the like.
“Cyclic” and “cycloalkyl” refer to a non-aromatic mono- or multicyclic ring system of about 3 to about 10 carbon atoms, e.g., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The cycloalkyl group can be optionally partially unsaturated. The cycloalkyl group also can be optionally substituted with an alkyl group substituent as defined herein, oxo, and/or alkylene. There can be optionally inserted along the cyclic alkyl chain one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms, wherein the nitrogen substituent is hydrogen, alkyl, substituted alkyl, aryl, or substituted aryl, thus providing a heterocyclic group. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Multicyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl.
“Alkoxyl” refers to an alkyl-O— group wherein alkyl is as previously described. The term “alkoxyl” as used herein can refer to, for example, methoxyl, ethoxyl, propoxyl, isopropoxyl, butoxyl, f-butoxyl, and pentoxyl. The term “oxyalkyl” can be used interchangeably with “alkoxyl”. In some embodiments, the alkoxyl has 1, 2, 3, 4, or 5 carbons.
“Aralkyl” refers to an aryl-alkyl group wherein aryl and alkyl are as previously described, and included substituted aryl and substituted alkyl. Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.
“Alkylene” refers to a straight or branched bivalent aliphatic hydrocarbon group having from 1 to about 20 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. The alkylene group can be straight, branched or cyclic. The alkylene group also can be optionally unsaturated and/or substituted with one or more “alkyl group substituents.” There can be optionally inserted along the alkylene group one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms (also referred to herein as “alkylaminoalkyl”), wherein the nitrogen substituent is alkyl as previously described. Exemplary alkylene groups include methylene (—CH2—); ethylene (—CH2—CH2—); propylene (—(CH2)3—); cyclohexylene (—C6H10—); —CH═CH—CH═CH—; —CH═CH—CH2—; wherein each of q and r is independently an integer from 0 to about 20, e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, and R is hydrogen or lower alkyl; methylenedioxyl (—O—CH2—O—); and ethylenedioxyl (—O—(CH2)2—O—). An alkylene group can have about 2 to about 3 carbon atoms and can further have 6-20 carbons.
“Arylene” refers to a bivalent aryl group. An exemplary arylene is phenylene, which can have ring carbon atoms available for bonding in ortho, meta, or para positions with regard to each other, i.e., respectively. The arylene group can also be napthylene. The arylene group can be optionally substituted (a “substituted arylene”) with one or more “aryl group substituents” as defined herein, which can be the same or different.
“Aralkylene” refers to a bivalent group that contains both alkyl and aryl groups. For example, aralkylene groups can have two alkyl groups and an aryl group (i.e., -alkyl-aryl-alkyl-), one alkyl group and one aryl group (i.e., -alkyl-aryl-) or two aryl groups and one alkyl group (i.e., -aryl-alkyl-aryl-).
The term “amino” and “amine” refer to nitrogen-containing groups such as NR3, NH3, NHR2, and NH2R, wherein R can be alkyl, branched alkyl, cycloalkyl, aryl, alkylene, arylene, aralkylene. Thus, “amino” as used herein can refer to a primary amine, a secondary amine, or a tertiary amine. In some embodiments, one R of an amino group can be a cation stabilized diazeniumdiolate (i.e., NONO−X+).
The terms “cationic amine” and “quaternary amine” refer to an amino group having an additional (i.e., a fourth) group, for example a hydrogen or an alkyl group bonded to the nitrogen. Thus, cationic and quartemary amines carry a positive charge.
The term “alkylamine” refers to the -alkyl-NH2 group.
The term “carbonyl” refers to the —(C═O)— group.
The term “carboxyl” refers to the —COOH group and the term “carboxylate” refers to an anion formed from a carboxyl group, i.e., —COO−.
The terms “halo”, “halide”, or “halogen” as used herein refer to fluoro, chloro, bromo, and iodo groups.
The term “hydroxyl” and “hydroxy” refer to the —OH group.
The term “hydroxyalkyl” refers to an alkyl group substituted with an —OH group.
The term “mercapto” or “thio” refers to the —SH group. The term “silyl” refers to groups comprising silicon atoms (Si).
As used herein the term “alkoxysilane” refers to a compound comprising one, two, three, or four alkoxy groups bonded to a silicon atom. For example, tetraalkoxysilane refers to Si(OR)4, wherein R is alkyl. Each alkyl group can be the same or different. An “alkylsilane” refers to an alkoxysilane wherein one or more of the alkoxy groups has been replaced with an alkyl group. Thus, an alkylsilane comprises at least one alkyl-Si bond. The term “fluorinated silane” refers to an alkylsilane wherein one of the alkyl groups is substituted with one or more fluorine atoms. The term “cationic or anionic silane” refers to an alkylsilane wherein one of the alkyl groups is further substituted with an alkyl substituent that has a positive (i.e., cationic) or a negative (i.e. anionic) charge, or can become charged (i.e., is ionizable) in a particular environment (i.e., in vivo).
The term “silanol” refers to a Si—OH group.
Provided according to some embodiments of the invention are wound dressings that include a polymer matrix and nitric oxide (NO)-releasing polysiloxane macromolecules within and/or on the polymer matrix. The appropriate combination of polymer matrix and NO-releasing polysiloxane macromolecules may allow for a wound dressing that stably stores NO and may provide for controlled release of NO to the wound.
The Polymer Matrix
As used herein, the term “polymer matrix” is meant to encompass any natural or synthetic polymeric material that may retain at least some of the NO-releasing polysiloxane macromolecules therein or thereon. As such, the polymer matrix may be a homopolymer, heteropolymer, random copolymer, block copolymer, graft copolymer, mixture or blend of any suitable polymer(s), and it may be in any suitable physical form, such as a foam, film, woven or non-woven material, hydrogel, gel matrix, mixtures and blends thereof, and the like. As described in further detail below, the choice of polymeric matrix and its physico-chemical properties for a particular wound dressing may depend on factors such as the NO-releasing polysiloxane macromolecules within and/or on the polymer matrix, and the type of therapeutic action desired.
In some embodiments of the invention, the polymer matrix includes at least one of hydrophilic polyurethanes, hydrophilic polyacrylates, co-polymers of carboxymethylcellulose and acrylic acid, N-vinylpyrrolidone, poly(hydroxy acids), polyanhydrides, polyorthoesters, polyamides, polycarbonates, polyalkylenes (e.g., polyethylene and polypropylene), polyalkylene glycols (e.g., poly(ethylene glycol)), polyalkylene oxides (e.g., polyethylene oxide), polyalkylene terephthalates (e.g., polyethylene terephthalate), polyvinyl alcohols, polyvinyl ethers, polylvinyl esters, polyvinyl halides (e.g., poly(vinyl chloride)), polyvinylpyrrolidone, polysiloxanes, poly(vinyl acetates), polystyrenes, polyurethane copolymers, cellulose, derivatized celluloses, alginates, poly(acrylic acid), poly(acrylic acid) derivatives, acrylic acid copolymers, methacrylic acid, methacrylic acid derivatives, methacrylic acid copolymers, poly(butyric acid), poly(valeric acid), poly(lactide-co-caprolactone), copolymers thereof and blends thereof.
In some embodiments of the invention, the polymer matrix may include a superabsorbent polymer (SAP). A polymer is considered superabsorbent, as defined per IUPAC, as a polymer that can absorb and retain large amounts of water relative to its own mass. SAPs may absorb water more than 500 times their own weight and may swell more than 1000 times their original volume. Exemplary SAPs include sodium polyacrylate, the polyurethane Tecophilic® TG-T2000, polyacrylamide copolymer, ethylene maleic anhydride copolymer, cross-linked carboxy-methyl-cellulose, polyvinyl alcohol copolymers, and cross-linked polyethylene oxide.
In some embodiments of the invention, polymers that are relatively hydrophobic, as defined by a water uptake value less than 10% by weight, may be used. Any suitable hydrophobic polymer may be used. However, exemplary polymers that are relatively hydrophobic include aromatic polyurethanes, silicone rubber, polycaprolactone, polycarbonate, polyvinylchloride, polyethylene, poly-L-lactide, poly-DL-glycolide, polyether etherketone (PEEK), polyamide, polyimide and polyvinyl acetate.
In addition, in some embodiments of the invention, the polymer matrix is modified to reduce swelling of the polymer and therefore prevent macromolecule leaching (e.g., NO-releasing polysiloxane macromolecule migration from the polymer matrix to the wound bed). Such modifications may include crosslinking of the polymer chains. The polymer matrix may also be modified by reacting the polymer with additional reagents. For example, the polymer may be modified to add hydrophilic groups, such as anionic, cationic and/or zwitterionic moeities, or to add hydrophobic groups such as silicone moeities, to the polymer chain.
In some embodiments, the polymer matrix includes polymer foam. The term “polymer foam” is meant to encompass any natural or synthetic polymer that is present as a foam and that may retain at least some NO-releasing polysiloxane macromolecules therein or thereon. In some embodiments, the polymer foam has an average cell size in a range of 50 μm and 600 μm. Furthermore, in some embodiments, the foam may be an open-celled foam. In some embodiments, the open cell walls of the foam may include pores of average size smaller than 100 μm, with the individual pore sizes between 10 and 80 μm. As used herein, the term “open-celled foam” refers to a foam that has cells that are substantially interconnected, such as foams wherein at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the cells are connected to at least one other cell. In some embodiments, the foam may be flexible. As used herein, the term “flexible” refers to foam that has a flexural strength of at least 40 MPa.
In some embodiments of the invention, the polymer foam is polyurethane foam. Any suitable polyurethane foam may be used. However, in some embodiments, the polyurethane foam may include at least one polyisocyanate segment and at least one polyol segment. Polyurethanes may be formed from the reaction of polyisocyanates and polyols. The polyisocyanate segment refers to a portion of the polyurethane formed from at least one polyisocyanate.
In some embodiments of the invention, the at least one polyisocyanate segment is formed from at least one of tolylene diisocyanate, methylphenylene diisocyanate, modified diisocyanates (e.g., uretdiones, isocyanurates, allophanates, biurets, isocyanate prepolymers and carbodiimide-modified isocyanates) and/or mixtures thereof. Exemplary diisocyanate include toluene diisocyanate; 1,4-tetramethylene diisocyanate; 1,4-hexamethylene diisocyanate; 1,6-hexamethylene diisocyanate; 1,12-dodecane diisocyante; cyclobutane-1,3-diisocyanate; cyclohexane-1,3-diisocyanate; cyclohexane-1,4-diisocyanate; 1-isocyanato-3,3,5-trimethyl-5-isocyanatomethyl cyclohexane; 2,4-hexahydrotolylene diisocyanate; 2,6-hexahydrotolylene diisocyanate; 2,6-hexahydro-1,3-phenylene diisocyanate; 2,6-hexahydro-1,4-phenylene diisocyanate; per-hydro-2,4′-diphenyl methane diisocyanate; per-hydro-4,4′-diphenyl methane diisocyanate; 1,3-phenylene diisocyanate; 1,4-phenylene diisocyanate; 2,4-tolylene diisocyanate, 2,6-toluene diisocyanates; diphenyl methane-2,4′-diisocyanate; diphenyl methane-4,4′-diisocyanate; naphthalene-1,5-diisocyanate; 1,3-xylylene diisocyanate; 1,4-xylylene diisocyanate; 4,4′-methylene-bis(cyclohexyl isocyanate); 4,4′-isopropyl-bis-(cyclohexyl isocyanate); 1,4-cyclohexyl diisocyanate; 3-isocyanatomethyl-3,5,5-trimethylcyclohexyl isocyanate (IPDI); 1-methyoxy-2,4-phenylene diisocyanate; 1-chloropyhenyl-2,4-diisocyante; p-(1-isocyanatoethyl)-phenyl isocyanate; m-(3-isocyanatobutyl)-phenyl isocyanate; 4-(2-isocyanate-cyclohexyl-methyl)-phenyl isocyanate; and mixtures thereof.
The polyol segment refers to a portion of the polyurethane foam formed from at least one polyol. The polyols may include polyether polyols and/or polyester polyols. Polyether polyols may have a significant amount of ether linkages in their structure, whereas the polyester polyols may have ester linkages within their structure. Any suitable polyol may be used. However, in some embodiments of the invention, the at least one polyol segment is formed from a diol having from 2 to 18 carbon atoms, and in some embodiments, a diol having 2 to 10 carbon atoms. Exemplary diols include 1,2-ethanediol, 1,3-propanediol, 1,4-butanediol, 1,6-hexanediol, 1,5-pentanediol, 1,10-decanediol, 2-methyl-1,3-propanediol, 2-methyl-2-butyl-1,3-propanediol, 2,2-dimethyl-1,3-propanediol, 2,2-dimethyl-1,4-butanediol, 2-ethyl-2-butyl-1,3-propanediol, neopentyl glycol hydroxypivalate, diethylene glycol and triethylene glycol. Triols and polyols of higher functionality may also be used and include compounds having 3 to 25, and in some embodiments, 3 to 18, and, in particular embodiments, 3 to 6 carbon atoms. Examples of triols which can be used are glycerol or trimethylolpropane. As polyols of higher functionality, it is possible, for example, to employ erythritol, pentaerythritol and sorbitol. In some embodiments, low molecular mass reaction products of the polyols may be used, for example, those of trimethylolpropane with alkylene oxides, such as ethylene oxide and/or propylene oxide. These low molecular mass polyols can be used individually, or as mixtures.
Examples of polyether polyols include the commercially available polyols PETOL28-3B, PETOL36-3BR, and PETOL56-3MB, Multranol®, Arcol® and Acclaim® (Bayer Material Science), and the Caradol® family of polyols (Shell Chemical Corporation).
Examples of polyester polyols that can be suitably used in the current invention include diethylene glycol adipate diol, dicaprylate diol, and in general, the esters of dicarboxylic and hydroxyl acids with glycol, and glycol functionalized polyester oligomers polylactate, polglycolate, polycaprylate, PET, and commercial formulations such as Desmophen® C polycarbonate diols and Desmophen® polyacrylate diols (Bayer).
In some embodiments, certain polyols or mixtures thereof specifically formulated for manufacture of high resiliency flexible polyurethane foams may be used. Examples of such polyols include a polyol having an ethylene oxide content in a range of 50% to 80%, a primary hydroxyl content of at least 40% and a molecular weight in a range of 2500 and 6000; a polymer polyol prepared by in situ polymerization of a high-functionality poly(oxyethylene)-poly(oxypropylene) oligomer, with a second poly(oxyethylene) polyol oligomer with a molecular weight ranging from 450 to 30,000 and an poly(oxyethylene) content greater than 70%; and biobased polyols derived from soybean oil, castor oil, palm oil, linseed oil and canola oil.
Other polyisocyanates and polyols may be used, including polyols and polyisocyanates having other functional groups therein. As such, in some embodiments, the polyisocyanates and polyols may include other functional groups such as ether, ester, urea, acrylate, pyrrolidone, vinyl, phenyl, and amino linkages, provided that the resulting polyurethane is suitable for forming a foam.
In some embodiments of the invention, the polymer foam includes a superabsorbent polymer (SAP). The superabsorbancy can be introduced in the foam structure by the inclusion of polymer segments that have superabsorbency in the polyol or the ‘soft’ segment of the foam. Examples of polymer segments having superabsorbency may include polyacrylamide copolymer, ethylene maleic anhydride copolymer, cross-linked carboxy-methyl-cellulose, polyvinyl alcohol copolymers, and cross-linked polyethylene oxide.
In some embodiments of the invention, polymer foams that are relatively hydrophobic may be used. Any suitable hydrophobic polymer may be used. Hydrophobicity can be introduced by selection of the polyisocyanate or the ‘soft’ segment of the polyurethane foam. Choosing highly hydrophobic polyisocyanates may results in a rigid foam, while lack of adequate hydrophobicity may prevent the formation of the foam structure.
Commonly used polyisocyanates for hydrophobic foams include diphenylmethane diisocyanate and its isomers, toluene diisocyanate, hexamethylene diisocyanate and mixtures thereof. In some embodiments, the polyisocyanates may also include copolymers of the diisocyanates previously mentioned. In some embodiments, the isocyanate groups may also be introduced at the termini of polymer segments of relatively hydrophobic polymers to provide a better control of the foam. Other polymers that are relatively hydrophobic include silicone rubber, polycaprolactone, polycarbonate, polyvinylchloride, polyethylene, poly-L-lactide, poly-DL-glycolide, polyetheretherketone (PEEK), polyamide, polyimide and polyvinyl acetate.
The polymer to be foamed may also be modified by reacting the polymer with additional reagents. For example, the polymer may be modified to add hydrophilic groups, such as anionic, cationic and/or zwitterionic moeities, or to add hydrophobic groups, such as silicone groups, to the polymer chain.
The polymer foam may be prepared by the reaction between the isocyanate moieties of the polyisocyanate ‘hard’ segments and the nucleophilic terminal groups of the polyols or ‘soft’ segments. The nucleophilic groups may include hydroxyl, amine and/or carboxylic groups.
In some embodiments, the foam may also contain chain extending segments in addition to the polyols and polyisocyanate building blocks. Polyamine co-reactants, due to the their reactivity with isocyanates, are the most commonly used chain extenders used to increase the chain length and flexibility of the foam. The most commonly used polyamines are polyaspartic esters, polyaldimines, butylenes diamines, and other short chain alkyl diamines.
Nitric Oxide-Releasing Polysiloxane Macromolecules
The term “NO-releasing polysiloxane macromolecules” refers to a structure synthesized from monomeric silane constituents that results in a larger molecular framework with a molar mass of at least 500 Da and a nominal diameter ranging from 0.1 nm-100 μm and may comprise the aggregation of two or more macromolecules, whereby the macromolecular structure is further modified with an NO donor group. For example, in some embodiments, the NO donor group may include diazeniumdiolate nitric oxide functional groups. In some embodiments, the NO donor group may include S-nitrosothiol functional groups.
In some embodiments of the invention, the NO-releasing polysiloxane macromolecules may be in the form of NO-releasing particles, such as those described in U.S. Publication No. 2009/0214618, the disclosure of which is incorporated by reference herein in its entirety. Such particles may be prepared by methods described therein.
As an example, in some embodiments of the invention, the NO-releasing particles include NO-loaded precipitated silica ranging from 20 nm to 10 μm in size. The NO-loaded precipitated silica may be formed from nitric oxide donor modified silane monomers into a co-condensed siloxane network. In some embodiments of the invention, the nitric oxide donor is an N-diazeniumdiolate.
In some embodiments of the invention, the nitric oxide donor may be formed from an aminoalkoxysilane by a pre-charging method, and the co-condensed siloxane network may be synthesized from the condensation of a silane mixture that includes an alkoxysilane and the aminoalkoxysilane to form a nitric oxide donor modified co-condensed siloxane network. As used herein, the “pre-charging method” means that aminoalkoxysilane is “pretreated” or “precharged” with nitric oxide prior to the co-condensation with alkoxysilane. In some embodiments, the precharging nitric oxide may be accomplished by chemical methods. In another embodiment, the “pre-charging” method can be used to create co-condensed siloxane networks and materials more densely functionalized with NO-donors.
The co-condensed siloxane network can be silica particles with a uniformed size, a collection of silica particles with a variety of size, amorphous silica, a fumed silica, a nanocrystalline silica, ceramic silica, colloidal silica, a silica coating, a silica film, organically modified silica, mesoporous silica, silica gel, bioactive glass, or any suitable form or state of silica.
The composition of the siloxane network, (e.g., amount or the chemical composition of the aminoalkoxysilane) and the nitric oxide charging conditions (e.g., the solvent and base) may be varied to optimize the amount and duration of nitric oxide release. Thus, in some embodiments, the composition of the silica particles may be modified to regulate the half-life of NO release from silica particles.
In some embodiments, the alkoxysilane is a tetraalkoxysilane having the formula Si(OR)4, wherein R is an alkyl group. The R groups can be the same or different. In some embodiments the tetraalkoxysilane is selected as tetramethyl orthosilicate (TMOS) or tetraethyl orthosilicate (TEOS). In some embodiments, the aminoalkoxysilane has the formula: R″—(NH—R′)n—Si(OR)3, wherein R is alkyl, R′ is alkylene, branched alkylene, or aralkylene, n is 1 or 2, and R″ is selected from the group consisting of alkyl, cycloalkyl, aryl, and alkylamine.
In some embodiments, the aminoalkoxysilane can be selected from N-(6-aminohexyl)aminopropyltrimethoxysilane (AHAP3); N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (AEAP3); (3-trimethoxysilylpropyl)di-ethylenetriamine (DET3); (amino ethylaminomethyl)phenethyltrimethoxysilane (AEMP3); [3-(methylamino)propyl]trimethoxysilane (MAP3); N-butylamino-propyltrimethoxysilane (n-BAP3); t-butylamino-propyltrimethoxysilane (t-BAP3); N-ethylaminoisobutyltrimethoxysilane (EAiB3); N-phenylamino-propyltrimethoxysilane (PAP3); and N-cyclohexylaminopropyltrimethoxysilane (cHAP3).
In some embodiments, the aminoalkoxysilane has the formula: NH[R′—Si(OR)3]2, wherein R is alkyl and R′ is alkylene. In some embodiments, the aminoalkoxysilane can be selected from bis(3-triethoxysilylpropyl)amine, bis-[3-(trimethoxysilyl)propyl]amine and bis-[(3-trimethoxysilyl)propyl]ethylenediamine.
In some embodiments, as described herein above, the aminoalkoxysilane is precharged for NO-release and the amino group is substituted by a diazeniumdiolate. Therefore, in some embodiments, the aminoalkoxysilane has the formula: R″—N(NONO−X+)—R′—Si(OR)3, wherein R is alkyl, R′ is alkylene or aralkylene, R″ is alkyl or alkylamine, and X+ is a cation selected from the group consisting of Na+, K+ and Li+.
In some embodiments of the invention, the co-condensed siloxane network further includes at least one crosslinkable functional moiety of formula (R1)x(R2)ySiR3, wherein R1 and R2 is each independently C1-5 alkyl or C1-5 alkoxyl, X and Y is each independently 0, 1, 2, or 3, and X+Y equal to 3, and R3 is a crosslinkable functional group. In a further embodiment, R1 is C1-3 alkoxyl, and R2 is methyl. In another embodiment, R3 is selected from the group consisting of acrylo, alkoxy, epoxy, hydroxy, mercapto, amino, isocyano, carboxy, vinyl and urea. R3 imparts an additional functionality to the silica which results in a multifunctional device. Yet, in another embodiment, the crosslinkable functional moiety is selected from the group consisting of methacryloxymethyltrimethoxysilane, methacryloxypropyltrimethoxysilane, methacryloxypropyltriethoxysilane, 3-acryloxypropyl)trimethoxysilane, N-(3-methyacryloxy-2-hydroxypropyl)-3-aminopropyltriethoxysilane, 5,6-epoxyhexyltriethoxysilane, 2-(3,4-epoxycyclohexyl)ethyl trimethoxysilane, 3-glycidoxypropyl)trimethoxysilane, 3-aminopropyltrimethoxysilane, 3-aminopropyltriethoxysilane, 3-isocyanatopropyltriethoxysilane, 3-mercaptopropyltrimethoxysilane, mercaptopropyltriethoxysilane, 11-mercaptoundecyltrimethoxysilane, 2-cyanoethyltriethoxysilane, ureidopropyltriethoxysilane, ureidopropyltrimethoxysilane, vinylmethyldiethoxysilane, vinylmethyldimethoxysilane, vinyltriethoxysilane, vinyltrimethoxysilane, vinyltriisopropoxysilane and vinyltris(2-methoxyethoxy)silane. In some embodiments, R3 may be used to cross-link the NO-donor modified silica with or within polymeric matrices.
The NO-releasing polysiloxane macromolecules may be present within and/or on the polymer matrix at any suitable concentration, but in some embodiments, the NO-releasing polysiloxane macromolecules are present within the polymer matrix at a concentration sufficient to increase the rate of wound healing, decrease inflammation and/or exert an antimicrobial effect. In particular embodiments, the concentration of the NO-releasing polysiloxane macromolecules may be in a range of about 0.1 to about 20 weight percent.
In some embodiments, the NO-releasing polysiloxane macromolecules may be uniformly distributed within the polymer matrix. Thus, in such embodiments, the concentration of NO-releasing polysiloxane macromolecules is substantially constant throughout the polymer matrix.
Interaction Between the Polymer Matrix and NO-Releasing Polysiloxane Macromolecules
Wound healing occurs in several different phases, and may take place over 0-12 (or more) months. Wound healing phases include:
The phase of wound healing plays a role in the selection of the NO-releasing polysiloxane macromolecules and the polymer matrix chosen. Nitric oxide may play a role in wound healing by a number of different mechanisms. First, extended exposure to low concentrations of nitric oxide may promote wound healing whereby nitric oxide acts as a signaling molecule in a number of wound healing cascades. Additionally, nitric oxide may also play a role in mitigating inflammation following injury. Modulation of inflammatory cytokines and cells of the inflammatory response via nitric oxide may significantly alter the wound healing phases described above. Additionally, wound complications and pain may be significantly reduced with topical administration of nitric oxide as an anti-inflammatory agent. Furthermore, nitric oxide may act as a broad spectrum antimicrobial agent, particularly at relatively high concentrations. The antimicrobial effects of nitric oxide are broad ranging and different wound types may be colonized with different wound pathogens (e.g., gram negative bacteria, gram positive bacteria, fungi, etc.). Additionally, different pathogens may be more sensitive to nitric oxide than other pathogens. In some embodiments, nitric oxide may act as an antimicrobial agent by directly killing planktonic bacteria and other organisms; directly killing biofilm embedded bacteria and other organisms; indirectly killing microorganisms through nitrosative/oxidative stress; increasing drug permeability across microbial membranes; and/or preventing recurrence of infection or biofilm formation.
Therefore, in some embodiments, the nitric oxide released from a particular wound dressing may provide a particular therapeutic action, such as act as a signaling molecule in a wound healing cascade, act as an anti-inflammatory agent and/or act as an antimicrobial agent. The desired therapeutic action may determine which NO-releasing polysiloxane macromolecules and polymer matrix are used in a particular wound dressing. For example, two particular classes of nitric oxide donors are diazeniumdiolates and nitrosothiols. Both of these nitric oxide donors have at least one mechanism for the release of nitric oxide. Diazeniumdiolate may be triggered to release nitric oxide by exposure to water or another proton source, and an O2-protected diazeniumdiolate may be triggered to release nitric oxide by exposure to light, enzymatic action and/or pH adjustment. Nitrosothiols may be triggered to release nitric oxide via thermal and radiative processes, and/or via interaction with copper and other thiols (e.g., glutathione). Therefore, the mechanism of release of nitric oxide from the NO-releasing polysiloxane macromolecules may affect which polymer matrix is chosen.
Specifically, because different NO-releasing polysiloxane macromolecules may release nitric oxide by different mechanisms, the polymer matrix chosen should complement the particular NO-releasing macromolecule employed. Several properties of the polymer matrix may be tailored based on the NO-releasing polysiloxane macromolecules used and the desired therapeutic action of the wound dressing. Such properties include:
(i) Moisture Uptake/Retention
The rate of moisture uptake may be tunable to meet the requirements of the NO-release kinetics of the macromolecule in order to achieve the desired therapeutic action of the wound dressing. The equilibrium moisture retention can vary from 5 percent for certain aliphatic polymers to over 2000 percent for hydrogels and superabsorbent polymers. Thus, in some embodiments, the polymer matrix has a low equilibrium moisture retention in a range of 0 to 10 percent. In some embodiments, the polymer matrix has a moderate equilibrium moisture retention in a range of 10 to 100 percent. Further, in some embodiments, the polymer matrix has a high equilibrium moisture retention of 100 percent or higher.
(ii) Moisture Vapor Transfer Rate (MVTR)
The MVTR may be tunable in breathable polymer films to match the requirements of a water reactive NO-releasing polysiloxane macromolecules in a thin film yet still maintain adequate MVTR for the desired wound or injury area. Wound dressings that maintain a moist wound bed are termed as occlusive. An optimum MVTR maintains a moist wound environment which activates debriding enzymes and growth factors that promote wound healing. Occlusive dressings also act as a barrier towards exogenous microbes, thereby preventing infection. Occlusive dressings are characterized by an MVTR of less than 35 g water/m2/h
(iii) Ability to Swell
The ability of the wound dressing to swell without dissolution upon contact with wound moisture is beneficial in highly exudating wounds. The wound dressing serves to imbibe excess moisture that may otherwise cause wound maceration and foul odor.
(iv) Surface Energy
Hydrophobic wound dressings are characterized by low surface energy whereas charged and/or hydrophilic wound dressings have a high surface energy. Low surface energy is desirable to allow easy removal of the dressing without damaging the wound bed.
(v) Oxygen Permeability
Adequate oxygen level facilitates neovascularization, aids in collagen synthesis, and may prevent or minimize microbial infection of the wound. Due to damaged vasculature in wounds, there may be a low oxygen tension in the wound bed, leading to hypoxia and anaerobic metabolism that can delay the healing process. Wound dressings may be oxygen permeable so that the wound receives adequate topical oxygen for healing.
(vi) Nitric Oxide Permeability
The polymer matrix of the wound dressing may have adequate permeability towards nitric oxide such that the nitric oxide generated by the NO-releasing polysiloxane macromolecules is available to the wound bed at a desired therapeutic rate. Hydrophilic materials typically have a lower NO permeability towards nitric oxide as compared to hydrophobic materials. The NO permeability of the dressing may be matched to the release kinetics of the NO-releasing polysiloxane macromolecules and the rate of water uptake by the polymer, in order to provide for optimal release of NO from the dressing.
(vii) Biodegradability/Bioabsorbability
Biodegradability refers to the property of the wound dressing to break down into smaller molecular weight components under physiological conditions. Bioresorbability refers to the property by which the wound dressing can break down into smaller molecular weight segments and the segments are completely taken into the body without any biological reaction. This property is desirable if the dressing is to be used over a long-term for cavity-type wounds.
(viii) Tensile Strength
Tensile strength is the ability of the wound dressing to withstand breakage upon elongation in any direction. The wound dressing material needs to have adequate tensile strength in order to withstand stresses occurring as a result of normal patient wear.
(ix) Biocompatibility
The polymer matrix of the wound dressing may be biocompatible, non-toxic, and non-irritable.
(x) Ionic Character
The ionic character of the dressing may affect the dressing surface energy and biocompatibility. The ionic character of the dressing can be quantified by measurement of the zeta potential of the wound dressing material under physiological conditions. In some embodiments, the zeta potential of surfaces may be between −30 mV and +20 mV, and in some embodiments, between −10 mV and +10 mV, and in some embodiments, approximately zero. Surfaces with highly negative (<−30 mV) or highly positive (>+20 mV) zeta potential may be undesirable as they may have an anti- or pro-coagulant effect on the wound and may increase dressing surface energy.
(xi) Transparency
The ability of the wound dressing material to allow passage of visible light may allow for visual monitoring of the wound healing process. As used herein, a wound dressing or polymer matrix is transparent if it has an optical transparency value of 80 percent or more transmittance as measured via solid state spectrophotometry.
As moisture facilitates nitric oxide release from diazeniumdiolate-functionalized polysiloxane macromolecules, a wound dressing that includes diazeniumdiolate-modified polysiloxane macromolecules within and/or on a hydrophilic polymer will allow for the release of nitric oxide to a wound at a greater rate than would a hydrophobic polymer. Thus, the level of nitric oxide desired to be applied to the wound can be tailored by increasing or decreasing the hydrophilicity of the polymer. Therefore, by combining a hydrophilic polymer with a diazeniumdiolate-modified macromolecule, a concentrated dose of nitric oxide may be provided to a wound, and by combining a diazeniumdiolate-modified macromolecule with a relatively hydrophobic polymer, an “extended release” dose of nitric oxide may be provided to the wound. An extended release formulation may allow for release of nitric oxide over a predetermined time, such as 0-7 days. Additionally, as thermal and/or light energy may facilitate nitrosothiol modified polysiloxane macromolecule decomposition, the polymer matrix and/or additional layers above the polymer matrix including the nitrosothiol-modified polysiloxane macromolecules may be transparent so that light may facilitate nitric oxide release from the nitrosothiol. The transparency may be modified to control the level of nitric oxide release.
Thus, the polymer matrix and the NO-releasing polysiloxane macromolecules may be selected based on at least one property of the polymer matrix and at least one property of the NO-releasing polysiloxane macromolecules such that the interaction of the properties of the polymer matrix and the NO-releasing polysiloxane macromolecules provides a predetermined characteristic to the wound dressing. In some embodiments of the invention, the at least one property of the polymer matrix may include moisture uptake/retention, moisture vapor transfer rate (MVTR), surface energy, oxygen permeability, nitric oxide permeability, pore size biodegradability/bioabsorbability, tensile strength, biocompatibility, ionic character and/or transparency. In some embodiments of the invention, the at least one property of the NO-releasing polysiloxane macromolecules may include the nitric oxide release mechanism (e.g., water, heat, light, enzymatic, pH, and the like), total quantity of nitric oxide stored in moles NO/mg silica, the hydrophobicity/hydrophilicity of the co-condensed silica, and the biodegradability/bioresorbability of the macromolecular framework. The predetermined characteristic may be the ability of the nitric oxide in the wound dressing to signal one or more wound healing cascades, to act as an anti-inflammatory agent and/or to act as an antimicrobial agent.
As used herein, the term “interaction of the properties” refers to the ability of particular properties of the polymer matrix and particular properties to the NO-releasing polysiloxane macromolecules to combine to produce a wound dressing that has a predetermined characteristic, as defined herein. For example, the particular hydrophilicity of the polymer matrix may interact with a particular concentration of water reactive NO-releasing polysiloxane macromolecules to produce the desired release rate of nitric oxide from the polymer matrix.
An exemplary calculation of the reaction rate of the nitric oxide release as a function of the rate of a dressing's water uptake, is shown below. In this calculation, the absorption of water over time in the wound dressing is modeled. It is assumed that the release of NO from the NO-releasing polysiloxane macromolecules commences immediately upon contact with the diffused water.
Referring to
wherein D=diffusion coefficient of water in the polymer matrix, C(t)=concentration of water at time t, and C∞ is the concentration of water at equilibrium, which is the surrounding concentration (55 M). In a given time, t, the water diffuses only up to a certain thickness into the polymer film, thereby ‘activating’ the NO-releasing polysiloxane macromolecules up to that depth, z(t). The concentration of water in the film is the concentration of water only up to this depth and is equal to the mass of water diffused up to that depth, divided by volume of the penetration, thus
where ϵ is the porosity of the polymer film, which is considered because the water will only penetrate into the interconnecting pores between the polymer chains. The mass of water in the film at given time (t) can be calculated by measuring the rate of water uptake (U) by the bulk polymer, which can be determined experimentally, and is defined as:
where Mpoly=mass of the polymer film. Thus substituting this expression for the mass of water, in the expression for depth of penetration as defined above,
If the intrinsic rate of NO release of the NO-releasing polysiloxane macromolecules is known as a function of time, ƒ(t), the rate of NO release from the silica in the polymer that have been ‘activated’ at time t is given by:
NO(t)=mNO(t)׃(t)
Where [NO(t)]=micromoles of NO released at time t, mNO=mass of NO-releasing silica activated at time (t). By definition the NO-releasing co-condensed silica loading in the polymer wound dressing is expressed as,
where Mpoly=mass of the entire polymer film, of which only Mpoly(t) has been ‘wet’ by the water at time (t). Therefore,
mNO(t)=0.01GMpoly(t)
mNO(t)=0.01GAρpolyz(t)
and hence,
Therefore, the overall rate of NO release from the polymer is given by
In some embodiments of the invention, the storage of nitric oxide in the dressing is in a range of 0.1 pmol NO cm−2 to 100 pmol NO cm−2. In some embodiments, the storage of nitric oxide release in the dressing is in a range of 10 pmol NO cm−2 to 1 nmol NO cm−2. In some embodiments, the storage of nitric oxide in the dressing is in a range of 1 nmol NO cm−2 to 10 μmol NO cm−2. Total nitric oxide storage (t[NO]) and surface flux can be measured in real-time via the chemiluminescent detection of nitric oxide reviewed by Hetrick et al. (Hetrick et al. Analytical Chemistry of Nitric Oxide, Annu. Rev. Anal. Chem. 2009, 2, 409-433, which is hereby incorporated by reference in its entirety). Additional kinetic parameters for nitric oxide release that can be measured during this technique are the time to release the maximum flux of NO (tm), quantity of NO at the maximum flux ([NO]m), half-life of nitric oxide release (t1/2), and nitric oxide release duration (td).
In some embodiments of the invention, the instantaneous flux of nitric oxide release from the hydrated dressing surface is in a range of 0.1 pmol NO cm−2 s−1 to 100 pmol NO cm−2 s−1 and constitutes a slow initial rate of release. In some embodiments, the instantaneous flux of nitric oxide release from the hydrated dressing surface is in a range of 100 pmol NO cm−2 s−1 to 1000 pmol NO cm−2 s−1 and constitutes an intermediate rate of release. In some embodiments, the instantaneous flux of nitric oxide from the hydrated dressing surface is in a range of 1 nmol NO cm−2 s−1 to 10 μmol NO cm−2 s−1 and constitutes a rapid burst or fast NO-release kinetics.
Stability
According to some embodiments of the invention, the wound dressings can stably store the NO so that NO is not released prior to its intended therapeutic use. In some embodiments, 95 percent or more of the original NO loading is retained after one week at 25° C. Furthermore, in some embodiments, 85 percent of the NO loading is retained up to 2 years at 25° C.
In some embodiments of the invention, the wound dressings form a stable matrix whereby the leaching of silica particles is minimized. The thermodynamics of particulate leaching from polymeric matrices has not been a challenge previously encountered in the prior art. Leaching of siloxane based macromolecules can be determined via static light scattering or elemental analysis for Si in the soak solutions. In some embodiments, greater than 98 percent of the embedded NO-releasing polysiloxane macromolecules is retained following incubation under physiological conditions (pH=7.4, 37° C., phosphate buffered saline) for 48 hours. In other embodiments, greater than 95 percent of the embedded NO-releasing polysiloxane macromolecules is retained following incubation under physiological conditions (pH=7.4, 37° C., phosphate buffered saline) for greater than 30 days.
Example Embodiments
In some embodiments, the NO-releasing polysiloxane macromolecule is Nitricil™ (Novan, Inc.), which is a diazeniumdiolate-modified precipitated silica.
In some embodiments, the polymer matrix is an aliphatic polyether polyurethane that absorbs water in an amount of about 6 percent to about 100 percent of its dry weight. In some embodiments, the aliphatic polyether polyurethane absorbs water in an amount of about 10 to 60 percent of its dry weight, and in some embodiments, 10 to 20 percent of its dry weight.
In some embodiments, the polymer matrix is a superabsorbent polymer that absorbs water in an amount of at least 100 percent and ranging up to 5000 percent of its dry weight. In some embodiments, the polymer matrix includes Tecophilic® Aliphatic thermoplastic polyurethane from Lubrizol, Inc.
Additives
In addition to the NO-releasing polysiloxane macromolecules, other additives may be present within and/or on the polymer matrix. Such additives may alter the properties of the polymeric matrix. For example, in some embodiments, the wound dressing may further include a water-soluble porogen. The water-soluble porogen is an additive that may facilitate water uptake and diffusion in the polymer matrix. Any suitable porogen may be used, but in some embodiments, the porogen may include sodium chloride, sucrose, glucose, lactose, sorbitol, xylitol, polyethylene glycol, polyvinylpyrrollidone, polyvinyl alcohol and mixtures thereof.
The properties of the polymer matrix and water uptake may also affect the release of nitric oxide, and so additives that affect the properties of the polymer matrix and/or water uptake may in turn affect the rate of release of nitric oxide from the NO-releasing polysiloxane macromolecules.
Additives may also be included in the polymer foam that directly affect the release of nitric oxide from the NO-releasing macromolecules. For example, in some embodiments, basic or other anionic species may be used to buffer the pH of the polymer foam to slow diazeniumdiolate decomposition and resulting nitric oxide release. In other embodiments, chelating agents may be used to scavenge metal ions like Fe2+, Cu2+ and Cu+ to preserve nitrosothiol NO donor stability and prevent rapid nitric oxide release. In some embodiments, additives may be added to enhance NO donor decomposition given the inherently slow NO-release kinetics of the NO-releasing polysiloxane macromolecule. For example, acidic or carboxylic acid functionalized additives may be added to the foam to create a low internal foam pH upon hydration and accelerate the decomposition of N-diazeniumdiolate donors. In another embodiment, cysteine or glutathione may be impregnated into the foam matrix to facilitate transnitrosation and subsequent thiol mediated decomposition of nitrosothiol containing macromolecules.
In addition, other additives useful for foam formation and processing may be included. For example, surface-active agents may be added to enhance mixing, act as mold-release agents and/or to influence the final cellular structure of the foam. Furthermore, blowing agents, and byproducts therefrom, may also be present within the polymer foam. Blowing agents are described in further detail below. Additives that may be useful in forming foams include surface-active agents to enhance mixing as well as to influence the final foam structure, and mold-release agents. Examplary surface active agents may be found in U.S. Pat. No. 6,316,662, the disclosure of which is incorporated herein in its entirety.
Additives may be present in the polymer matrix that may act to provide additional therapeutic effects to the wound dressing, either acting synergistically or separately from the NO-releasing polysiloxane macromolecules. For example, in some embodiments, the wound dressings may also include at least one therapeutic agent such as antimicrobial agents, anti-inflammatory agents, analgesic agents, anesthetic agents, antihistamine agents, antiseptic agents, immunosuppressants, antihemorrhagic agents, vasodilators, wound healing agents, anti-biofilm agents and mixtures thereof.
Examples of antimicrobial agents include penicillins and related drugs, carbapenems, cephalosporins and related drugs, erythromycin, aminoglycosides, bacitracin, gramicidin, mupirocin, chloramphenicol, thiamphenicol, fusidate sodium, lincomycin, clindamycin, macrolides, novobiocin, polymyxins, rifamycins, spectinomysin, tetracyclines, vanomycin, teicoplanin, streptogramins, anti-folate agents including sulfonamides, trimethoprim and its combinations and pyrimethamine, synthetic antibacterials including nitrofurans, methenamine mandelate and methenamine hippurate, nitroimidazoles, quinolones, fluoroquinolones, isoniazid, ethambutol, pyrazinamide, para-aminosalicylic acid (PAS), cycloserine, capreomycin, ethionamide, prothionamide, thiacetazone, viomycin, eveminomycin, glycopeptide, glyclyclycline, ketolides, oxazolidinone; imipenen, amikacin, netilmicin, fosfomycin, gentamycin, ceftriaxone, Ziracin, Linezolid, Synercid, Aztreonam, and Metronidazole, Epiroprim, Sanfetrinem sodium, Biapenem, Dynemicin, Cefluprenam, Cefoselis, Sanfetrinem celexetil, Cefpirome, Mersacidin, Rifalazil, Kosan, Lenapenem, Veneprim, Sulopenem, ritipenam acoxyl, Cyclothialidine, micacocidin A, carumonam, Cefozopran and Cefetamet pivoxil. Examples of antihistamine agents include diphenhydramine hydrochloride, diphenhydramine salicylate, diphenhydramine, chlorpheniramine hydrochloride, chlorpheniramine maleate isothipendyl hydrochloride, tripelennamine hydrochloride, promethazine hydrochloride, methdilazine hydrochloride, and the like. Examples of local anesthetic agents include dibucaine hydrochloride, dibucaine, lidocaine hydrochloride, lidocaine, benzocaine, p-buthylaminobenzoic acid 2-(die-ethylamino) ethyl ester hydrochloride, procaine hydrochloride, tetracaine, tetracaine hydrochloride, chloroprocaine hydrochloride, oxyprocaine hydrochloride, mepivacaine, cocaine hydrochloride, piperocaine hydrochloride, dyclonine and dyclonine hydrochloride.
Examples of antiseptic agents include alcohols, quaternary ammonium compounds, boric acid, chlorhexidine and chlorhexidine derivatives, iodine, phenols, terpenes, bactericides, disinfectants including thimerosal, phenol, thymol, benzalkonium chloride, benzethonium chloride, chlorhexidine, povidone iode, cetylpyridinium chloride, eugenol and trimethylammonium bromide.
Examples of anti-inflammatory agents include nonsteroidal anti-inflammatory agents (NSAIDs); propionic acid derivatives such as, ibuprofen and naproxen; acetic acid derivatives such as indomethacin; enolic acid derivatives such as meloxicam, acetaminophen; methyl salicylate; monoglycol salicylate; aspirin; mefenamic acid; flufenamic acid; indomethacin; diclofenac; alclofenac; diclofenac sodium; ibuprofen; ketoprofen; naproxen; pranoprofen; fenoprofen; sulindac; fenclofenac; clidanac; flurbiprofen; fentiazac; bufexamac; piroxicam; phenylbutazone; oxyphenbutazone; clofezone; pentazocine; mepirizole; tiaramide hydrochloride; steroids such as clobetasol propionate, bethamethasone dipropionate, halbetasol proprionate, diflorasone diacetate, fluocinonide, halcinonide, amcinonide, desoximetasone, triamcinolone acetonide, mometasone furoate, fluticasone proprionate, betamethasone diproprionate, triamcinolone acetonide, fluticasone propionate, desonide, fluocinolone acetonide, hydrocortisone valerate, prednicarbate, triamcinolone acetonide, fluocinolone acetonide, hydrocortisone and others known in the art, predonisolone, dexamethasone, fluocinolone acetonide, hydrocortisone acetate, predonisolone acetate, methylpredonisolone, dexamethasone acetate, betamethasone, betamethasone valerate, flumetasone, fluorometholone, beclomethasone diproprionate, fluocinonide, topical corticosteroids, and may be one of the lower potency corticosteroids such as hydrocortisone, hydrocortisone-21-monoesters (e.g., hydrocortisone-21-acetate, hydrocortisone-21-butyrate, hydrocortisone-21-propionate, hydrocortisone-21-valerate, etc.), hydrocortisone-17,21-diesters (e.g., hydrocortisone-17,21-diacetate, hydrocortisone-17-acetate-21-butyrate, hydrocortisone-17,21-dibutyrate, etc.), alclometasone, dexamethasone, flumethasone, prednisolone, or methylprednisolone, or may be a higher potency corticosteroid such as clobetasol propionate, betamethasone benzoate, betamethasone dipropionate, diflorasone diacetate, fluocinonide, mometasone furoate, triamcinolone acetonide.
Examples of analgesic agents include alfentanil, benzocaine, buprenorphine, butorphanol, butamben, capsaicin, clonidine, codeine, dibucaine, enkephalin, fentanyl, hydrocodone, hydromorphone, indomethacin, lidocaine, levorphanol, meperidine, methadone, morphine, nicomorphine, opium, oxybuprocaine, oxycodone, oxymorphone, pentazocine, pramoxine, proparacaine, propoxyphene, proxymetacaine, sufentanil, tetracaine and tramadol.
Examples of anesthetic agents include alcohols such as phenol; benzyl benzoate; calamine; chloroxylenol; dyclonine; ketamine; menthol; pramoxine; resorcinol; troclosan; procaine drugs such as benzocaine, bupivacaine, chloroprocaine; cinchocaine; cocaine; dexivacaine; diamocaine; dibucaine; etidocaine; hexylcaine; levobupivacaine; lidocaine; mepivacaine; oxethazaine; prilocalne; procaine; proparacaine; propoxycaine; pyrrocaine; risocaine; rodocaine; ropivacaine; tetracaine; and derivatives, such as pharmaceutically acceptable salts and esters including bupivacaine HCl, chloroprocaine HCl, diamocaine cyclamate, dibucaine HCl, dyclonine HCl, etidocaine HCl, levobupivacaine HCl, lidocaine HCl, mepivacaine HCl, pramoxine HCl, prilocalne HCl, procaine HCl, proparacaine HCl, propoxycaine HCl, ropivacaine HCl, and tetracaine HCl.
Examples of antihemorrhagic agents include thrombin, phytonadione, protamine sulfate, aminocaproic acid, tranexamic acid, carbazochrome, carbaxochrome sodium sulfanate, rutin and hesperidin.
Wound Dressing Devices
Any suitable configuration of wound dressing device may be used. Referring to
Referring to
Referring to
The perforated layer 115 may be formed by any suitable method. However, in some embodiments, the perforated layer 115 is formed by using a mold, or by pressing an object into the polymer matrix 103 to form at least one hole 117, wherein the hole edges may also be melted and fused depending on the nature of the polymer matrix 103. Additionally, in some embodiments, the holes 117 of the perforated layers 115 may be formed by curing the polymer matrix 103 in a mold, e.g., in an anhydrous and/or low temperature process, and then peeled out or packaged in individual perforated trays.
According to some embodiments of the invention, additional therapeutic agents, such as those described herein, may be present in any of the layers of the wound dressing. As an example, in some embodiments, a layer that is substantially free of NO-releasing polysiloxane macromolecules may include at least one therapeutic agent. As an additional example, a layer that includes NO-releasing polysiloxane macromolecules may also include at least one additional therapeutic agent.
In some embodiments, the wound dressing may further include a polymer backing layer that contacts the polymer matrix or a polymer layer on the polymer matrix. In some embodiments, the wound dressing is an island wound dressing, and the polymer backing layer, and optionally at least a portion of the polymer matrix that contacts the wound bed, may include a medical grade adhesive thereon. For example, the wound dressing may include a polymer backing layer and a polymer matrix layer including a polymer matrix having NO-releasing polysiloxane macromolecules therein or thereon, wherein the polymer matrix layer is attached to a portion of the polymer backing layer, and wherein at least a portion of the polymer backing layer that is facing but not attached to the polymer matrix layer is coated with a medical grade adhesive. Such wound dressings may also include additional layers, such as a wound contact layer, wherein the wound contact layer is on the face of the polymer matrix layer that is not attached to the polymer backing layer.
Each layer of wound dressing according to embodiments of the invention may have any suitable thickness. However, in some embodiments, one or more layers of the wound dressing may have a thickness in a range of about 10 to about 5000 microns. In some embodiments of the invention, at least one layer of the wound dressing may be substantially transparent. Further, in some embodiments, the wound dressing as a whole may be substantially transparent. The term “substantially transparent” refers to a material that has a percent transmittance of 80 percent or more, as determined using a solid state spectrophotometer. Additionally, as described above, in some embodiments, at least one layer of the wound dressing has a medical-grade adhesive thereon. For example, the surface of the wound dressing that contacts the wound may have a medical-grade adhesive thereon.
Examples of medical grade adhesives that can be safely used on skin are acrylate-based adhesives, such as 2-ethylhexyl acrylate, isooctyl acrylate or n-butyl acrylate copolymerized with polar functional monomers such as acrylic acid, methacrylic acid, vinyl acetate, methyl acrylate, N-vinylcaprolactam, or hydroxyethyl methacrylate. Additional examples include octylcyanoacrylate, AcrySure™ adhesives (MACtac), adhesives based on silk protein, silicone gel based adhesives (Silbione® by Bluestar Silicones) and polyurethane based adhesive blends.
Wound Dressing Kits
As described above, wound healing may be effected through prolonged low concentrations of nitric oxide administration whereby nitric oxide acts as a signaling molecule in a number of wound healing cascades. In some embodiments, the instantaneous flux of nitric oxide release from hydrated dressing surface necessary to promote wound healing may be in the range of 0.5 pmol NO cm−2 s−1 to 20 pmol NO cm−2 s−1 upon initial application to the patient and constitutes a slow rate of release. An intermediate NO-releasing wound dressing may mitigate the inflammatory phase immediately following injury, following debridement of a chronic wound, in stalled wounds, or in infected wounds. In the inflammatory phase, the flux of nitric oxide released from the hydrated dressing surface is in a range of 20 pmol NO cm−2 s−1 to 1000 pmol NO cm−2 s−1 upon initial application to the patient and constitutes an intermediate rate of release. High levels of NO-released from a third matrix/NO-releasing polysiloxane macromolecule composition may be necessary to effect antimicrobial activity, using the rapid burst of nitric oxide to kill microorganisms through oxidative/nitrosative intermediates. In these embodiments, the flux of nitric oxide released from the hydrated dressing surface is in a range of 1 nmol NO cm−2 s−1 to 1 μmol NO cm−2 s−1 upon initial application and may constitute the rapid burst of nitric oxide necessary to provide a one or more log reduction against a broad range of microorganisms.
Therefore, provided according to some embodiments of the invention are kits that include wound dressings directed to a course of therapy with three unique dressing types of compositions designed to target these three wound processes. For a particular wound, a regiment may be implemented for a specified number of days whereby the three unique dressings are administered in sequence or repeated at some frequency (e.g., to keep microbial burden low).
Methods of Treating a Wound
In some embodiments of the invention, provided are methods of treating a wound by applying a wound dressing according to an embodiment of the invention. Such methods may be used in combination with any other known methods of wound treatment, including the application of medicaments, such as those that have anti-inflammatory, pain-relieving, immunosuppressant, vasodilating, wound healing and/or anti-biofilm forming properties. For the methods used herein, additional therapeutic agents and methods may be used prior to, concurrently with or after application with a gel according to embodiments of the invention. Wound dressings according to embodiments of the invention may also be used in combination with other wound dressings known to those of skill in the art.
In some embodiments of the invention, the wound dressings provided herein may be used in conjunction with at least one agent that can disrupt biofilm macrostructure prior to or in conjunction with the application of the wound dressing. In some embodiments, the anti-biofilm agent may disrupt the extracellular matrix of the biofilm. Examples of anti-biofilm agents that may act in this manner include lactoferrin, periodate, xylitol, DNase, protease, and an enzyme that degrades extracellular polysaccharides. In some embodiments of the invention, the formulation of the anti-biofilm agent may be acidic to promote enzyme activity of the DNase (e.g., mammalian DNases such as DNase II) and the acidic conditions simultaneously may also enhance the rate NO release from diazeniumdiolate modified silica. In some embodiments, the protease may include at least one of proteinase K, trypsin, Pectinex Ultra SP (PUS) and pancreatin. In some embodiments, enzymes that degrade extracellular polysaccharides may include N-acetylglucosaminidases (e.g., dispersin B).
In some embodiments of the invention, the anti-biofilm agent may act by affecting the transcriptional, translational and/or post-translational regulation of quorum-sensing genes or gene products in the infecting organism(s). For example, the anti-biofilm agents may include at least one of hamamelitannin, cyclic di-GMP and sublethal concentrations of nitric oxide.
The anti-biofilm agents may also act by other mechanisms. For example, the anti-biofilm agent may cause the infecting organism to transition from a sessile state to a metabolically active state. As another example, the anti-biofilm agent may act by causing the infecting organism(s) to transition from a non-motile state to a motile phenotype.
In some embodiments of the invention, the wound dressings provided herein may be used in conjunction with a wound debridement procedure. For example, in some embodiments, wounds may first be treated with a debridement procedure; and then a wound dressing according to an embodiment of the invention may be applied to the debrided wound. The wound dressings according to embodiments of the invention may increase the rate of wound healing, decrease inflammation and/or exert an antimicrobial effect. The wound dressings according to embodiments of the invention may be used in conjunction with any suitable debridement procedure. For example, the debridement procedure may be selective or nonselective.
In some embodiments, the debridement procedure may include at least one of surgical, enzymatic, autolytic, sharp, mechanical and biological processes. Any suitable surgical method may be used, but in some embodiments, the surgical method may involve a surgeon cutting away nonviable tissue in the wound. Any suitable enzymatic method may be used, but in some embodiments, the enzymatic method may involve the use of one or more proteases, their required cofactors, and optionally any enhancing agents, to digest the nonviable tissue in the wound. Exemplary proteases include trypsin, papain or other vegetable-derived proteases and collagenase. Any suitable autolytic method may be used, but in some embodiments, the autolytic method may involve maintaining a moist wound environment in order to promote the breakdown of nonviable tissue by enzymes that are naturally produced by the body. Any suitable mechanical method may be used, but in some embodiments, the mechanical methods may include wet-to-dry gauze, irrigation, pulsatile lavage, whirlpool therapy and/or low frequency ultrasound. Any suitable sharp method may be used, but in some embodiments, the sharp method may involve cutting away nonviable tissue by qualified clinical staff (e.g. RN or nurse practitioner). Any suitable biological method may be used, but in some embodiments, the biological method may involve the use of maggots, which selectively digest the nonviable tissue in the wound. These debridement methods may be used alone or in combination.
After the wound is debrided, a wound dressing according to an embodiment of the invention may be applied. Additional processes may be performed and therapeutic agents may be applied. For example, after wound debridement, an anti-biofilm agent may be applied to the wound prior to or in conjunction with the application of the wound dressing. Exemplary anti-biofilm agents include acetylsalicylic acid (aspirin), cyclic di-GMP, lactoferrin, gallium, selenium, as described above. Other compounds, such as hamamelitannin (witch hazel extract), arginine and c-di-GMP, may also be applied.
Also provided according to some embodiments of the invention are methods of using a wound dressing according to an embodiment of the invention in conjunction with negative pressure wound therapy (NPWT).
Subjects suitable to be treated with wound dressings or methods according to an embodiments of the invention include, but are not limited to, avian and mammalian subjects. Mammals of the present invention include, but are not limited to, canines, felines, bovines, caprines, equines, ovines, porcines, rodents (e.g. rats and mice), lagomorphs, primates, humans, and the like, and mammals in utero. Any mammalian subject in need of being treated according to the present invention is suitable. Human subjects are preferred. Human subjects of both genders and at any stage of development (i.e., neonate, infant, juvenile, adolescent, adult) can be treated according to the present invention.
Illustrative avians according to the present invention include chickens, ducks, turkeys, geese, quail, pheasant, ratites (e.g., ostrich) and, domesticated birds (e.g., parrots and canaries), and birds in ovo.
The invention can also be carried out on animal subjects, particularly mammalian subjects such as mice, rats, dogs, cats, livestock and horses for veterinary purposes, and for drug screening and drug development purposes.
Methods of Forming Wound Dressings
The wound dressings described herein may be formed by any suitable method. However, provided according to some embodiments of the invention are methods of forming wound dressings. In some embodiments, incorporation of NO-releasing polysiloxane macromolecules can be achieved through physically embedding the particles into polymer surfaces, via electrostatic association of particles onto polymeric surfaces, and/or by covalent attachment or cross-linking of particles onto reactive groups on the surface of a polymer, within the polymer and/or within cells of a foam. In some embodiments, methods of forming wound dressings include combining NO-releasing polysiloxane macromolecules and at least one monomer; and polymerizing the at least one monomer to form a polymer matrix comprising the NO-releasing polysiloxane macromolecules. The monomer may be polymerized by any suitable method, but in some embodiments, the monomer is polymerized by photocuring and/or moisture curing, with or without an initiator. In some embodiments, the monomer may be polymerized upon contact with the wound environment, e.g., via the moisture in the wound. In some embodiments, a single layer wound dressing may be formed by a method that includes solvent casting a solution of polymer and NO-releasing polysiloxane macromolecules.
In some embodiments, the polymerization occurs via liquid casting or molten polymer extrusion. In some embodiments, a liquid monomer, NO-releasing polysiloxane macromolecules and an initiator are deposited on a surface and polymerization proceeds upon activation of the initiator. Polymerizable groups can also be used to functionalize the exterior of the particles, whereupon, the particles can be co-polymerized into a polymer during the polymerization process.
In some embodiments of the invention, methods of forming the wound dressings include dispersing the NO-releasing polysiloxane macromolecules in a mixture of foam forming monomers; polymerizing the foam forming monomers to form a polymer; and then foaming the polymer.
In some embodiments, methods of forming wound dressings include reacting functional groups on the NO-releasing polysiloxane macromolecules with at least one foam forming monomer; polymerizing the at least one foam forming monomer to form a polymer including the NO-releasing polysiloxane macromolecules therein; and then foaming the polymer.
In some embodiments, polyurethane foam dressings may be prepared by the reaction of polyols with polyisocyanates added in stoichiometric excess, with other co-reactants added as required. In conventional foam manufacture, a stoichiometric amount of water is added to the reactant mix. The water may react with the isocyanate groups to form CO2 which bubbles through the polymerizing mass, creating a cellular structure of flexible foams.
For water reactive NO-releasing polysiloxane macromolecules, water may not be used in the preparation of NO-releasing foam dressings as water may activate the NO-releasing polysiloxane macromolecules, resulting in a premature release of NO and a decrease in the therapeutic value of the foam dressing.
Entirely non-aqueous foams may be synthesized by substituting or complementing the polyhydrols with amino alcohols or alkanolamines. Examplary amino alcohols and alkanolamines may be found in U.S. Pat. No. 5,859,285, the disclosure of which is incorporated herein in its entirety. The alkanolamines may chemically store CO2 on their amine groups, and this CO2 may be released by heating. The alkanolamines may be dissolved in a polar solvent, preferably a diol or a triol, and contacted with CO2 to form carbamates. In some embodiments, the polar solvent may be the polyol itself that can be the soft segment in the foam dressing.
The carbamate solution may be used to react with polyisocyanate to form the polyurethane foams. The carbamates may act as catalysts in foam formation, thereby avoiding the necessity of using other catalysts.
While any suitable alkanolamines may be used to produce carbamates, in some embodiments, the carbamates may be produced by using the following alkanolamines: 2-(2-aminoethylamino)ethanol, (3-[(2-aminoethyl)amino)]propanol), (2-[(3-aminopropyl)amino]ethanol), (1-[(2-aminoethyl)amino]-2-propanol, (2-[(3-aminopropyl)methylamino]ethanol, 1-[(2-amino-1-methylethyl)amino]-2-propanol, 2-[((2-amino-2-methylpropyl)amino]-2-methyl-1-propanol, 2-[(4-amino-3-methylbutyl)amino]-2-methyl-1-propanol, 17-amino-3,6,9,12,15-pentaazaheptadecan-1-ol and/or 3,7,12,16-tetraazaoctadecane-1,18-diol, in particular those based on 2-(2-aminoethylamino)ethanol as alkanolamine. The carbamate solution can be further blended with polyhydroxyl or polyamine containing compounds that have been pre-charged with NO and as a result, possess single or multiple NO-releasing functional group.
In addition to chemically storing the CO2 blowing agent in the above manner, physical blowing agents may also be used in foam production. Examples of physical blowing agents include: hydrohalo-olefin (See U.S. Patent Application Publication No. 20090099272, the contents of which are incorporated herein by reference in their entirety); alkanes, such as 2-methylbutane, pentane, heptanes (See U.S. Pat. No. 5,194,325, the contents of which are incorporated by reference in their entirety), and other inert, low-boiling compounds such as pentene and acetone. Carbon dioxide, including supercritical carbon dioxide, may be used as a physical blowing agent as well.
In some embodiments of the invention, provided are methods of forming multilayer wound dressings that include combining one or more polymeric layers, wherein one or more polymer layers may include a polymer matrix and NO-releasing polysiloxane macromolecules therein or thereon. The combining of polymeric layers may be achieved by any suitable method, but in some embodiments, the polymer layers are laminated to each other. Exemplary laminating techniques include ultrasonic welding, annealing with anhydrous organic solvents and application of a pressure sensitive adhesive.
The present invention will now be described in more detail with reference to the following examples. However, these examples are given for the purpose of illustration and are not to be construed as limiting the scope of the invention.
The water uptake for three hydrophilic polyurethanes thin film dressings soaked in phosphate buffered saline at physiological temperature and pH is shown in
The water uptake for Tecophilic® Aliphatic thermoplastic polyurethane HP-60D-20 (“T20”) loaded with increasing weight percent of poly(ethylene glycol) 8000 MW as a porogen is shown in
The water uptake for Tecophilic® Hydrogel thermoplastic polyurethane TG-2000 solvent cast into polymer films thin film dressings and soaked in phosphate buffered saline at physiological temperature and pH is shown in
P. aeruginosa
Upon application to the wound, the polyurethane material of the wound dressing comes in contact with moisture in the wound bed. The polyurethane has some affinity for moisture due to the presence of polyether soft-chain segments in its structure, which results in a finite amount of water being taken up by the dressing. This inward diffusion of moisture within the polymer matrix leads to an increase in distance between the polymer chains and is observed as swelling of the polymer. The increasing distance between the chains and the concentration gradient between the water in the polymer and the bulk water in the wound bed, allow greater space of movement for the embedded silica particles. As a result, the particles may diffuse out of the polymer, with the smaller particles having a greater propensity. This phenomenon manifests itself as particle leaching.
Wound dressings comprising NO-releasing silica particles were engineered to minimize leaching upon exposure to moisture in the wound bed. The dressing polymer composition and silica loading wt/wt both affect the cumulative amount leached. Light scattering is a commonly used technique to characterize particle suspensions, particularly to measure the size and polydispersity of micrometer and nanometer particle sizes. In static light scattering mode, the time-averaged intensity of light scattered by a particle suspension is measured and is highly dependent upon the particle size, its concentration, and molecular weight. Thus, for a dilute suspension of fairly monodisperse particles, the time averaged light intensity should be directly proportional to the particle concentration, and the static light scattering is said to occur in the Rayleigh mode. A plot of scattering intensity vs. particle concentration yields a straight line and provide an accurate method for determining unknown particle concentrations that leach into solution from wound dressing prototypes.
To measure the potential concentrations of NO-releasing silica that accumulate in solution following incubation under physiologically buffered solutions for 24 and 48 hr, the light scattering intensity of each unknown particle sample was measured and converted to its concentration using a calibration curve. Nitric oxide-releasing silica-loaded polyurethane dressings were cut into three 0.75″×0.75″ square samples. The samples were weighed and placed into polypropylene vials and 10 mL of filtered phosphate buffered saline (PBS, 10 mM sodium dihydrogen phosphate, 137 mM NaCl, 2.3 mM KCl, pH 7.4) pre-warmed at 37° C. was added to each. The vials were then incubated in a water bath maintained at 37° C. After 24 hours, each of the vials were agitated and an aliquot was removed, which was transferred to a polystyrene cuvette. The static light scattering intensity of this aliquot was determined against filtered PBS as blank. The silica concentration of the leachate was then determined using the calibration curve to convert the obtained kcps value to mg/ml, and the obtained mg/ml values were expressed as a percentage of the amount of silica calculated to be initially loaded in the dressing sample.
The leaching values for representative compositions are shown below and illustrate the dependence on polymer hydrophilicity and homogeneity of the silica polyurethane composite: 5% w/v T20 in tetrahydrofuran, 80 mg silica/g polymer, slurry prepared via magnetic stirring (TABLE 2), 5% w/v T20 in tetrahydrofuran, 80 mg silica/g polymer, slurry prepared via sonication, (TABLE 3), 10% w/v T100 in tetrahydrofuran, 160 mg silica/g polymer, slurry prepared via magnetic stirring (TABLE 4), 10% w/v T20 in tetrahydrofuran, 160 mg silica/g polymer, slurry prepared via sonication (TABLE 5). All wound dressing polymers were solvent cast into thin films and dried under vacuum.
P. aeruginosa biofilms were grown for 48 h in partial thickness wounds in a porcine animal model. After 2 days of growth, the baseline levels of bacteria from a flush of the wound with sterile buffer and a vigorous scrub of the wound with bacteria/tissue in sterile buffer were recorded. The planktonic bacteria were approximately 108 CFU/mL and the biofilm embedded bacteria were above 1010 CFU/mL prior to treatment with NO-releasing wound dressings. The efficacy of various NO-releasing wound dressing compositions on both the levels of planktonic bacteria flushed from the wound and the levels of biofilm bacteria scrubbed from the wound are shown in comparison to Tegaderm™ covered controls in
A medical grade, aliphatic polyether polyurethane that absorbs a small amount of water in the amount of 20% of its dry weight is combined with 14% w/w Nitricil™ 70 (nitric oxide-loaded precipitated silica) such that the Nitricil™ is permanently incorporated throughout the polyurethane matrix. The resulting polyurethane film device is transparent. The polymer blend is cast onto a transparent, siliconized PET release liner (FRA-308, Fox River Associates, LLC), which is pre-printed with a 1″ square grid.
The testing performed to evaluate design and technical characteristics is summarized in the TABLE 6 below.
Chemiluminescent detection of NO was used to characterize the nitric oxide release behavior from the polyurethane film device. The nitric oxide in the device is liberated upon exposure to moisture.
The nitric oxide-releasing wound dressing of Example 9 was used to treat partial thickness wounds in a porcine model. This study was designed to assess healing potential and whether or not the proposed device has a negative impact on normal wound repair in comparison to topical nitric oxide formulations previously reported (evidenced by delayed wound healing or significant erythema/edema). 160 rectangular wounds measuring 10 mm×7 mm×0.5 mm deep were divided into four treatment groups (40 wounds each). Wounds were dressed immediately after wounding and dressings were changed on days 2, 3, 5, and 7. Five wounds from each of the four groups were excised each day beginning on Day 4 after wounding and prepared according to the sodium bromide salt-split technique to assess epidermal migration. Epithelialization is considered complete (healed) if no defects or holes are present after the separation of the dermis and epidermis. Wounds in each of the groups were evaluated until 100% complete epithelialization was observed. The test materials were non-adherent to wound bed upon removal (no re-injury was observed) and none of the wounds from any of the treatment groups developed erythema, swelling or signs of infection. On Day 4, none of the treatment groups were completely re-epithelialized but Day 6, 100% of the wounds in the nitric oxide treated group were re-epithelialized in comparison to only 60% of the Tegaderm covered occlusive wound environment (
Testing has been performed by an independent laboratory, in accordance with Good Laboratory Practices, to evaluate the biocompatibility of the wound dressing in Example 9, as recommended by FDA's Blue Book Memo, G95-1, Use of International Standards ISO-10993, and Biological Evaluation of Medical Devices Part 1: Evaluation and Testing. Following are the tests that have been conducted along with a brief summary of results.
The wound dressing of Example 9 passed the requirements of all biocompatibility tests; thus it can be concluded that the product is biocompatible and non-toxic, providing a topical nitric oxide releasing solution with proven safety and effectiveness.
This example describes a process of manufacturing NO-releasing flexible polyurethane foam starting from 100 kg of 2-(2-aminoethylamino) ethanol as a basis. The foam is prepared using Desmodur N-100 (22% NCO groups, Bayer Material Science, Pittsburgh, Pa.) as the polyisocyanate and Desmodur N- and Desmophen-R-221-75 (3.3% OH groups, Bayer Material Science, Pittsburgh, Pa.) as the polyol. Nitricil™-70 (Novan, Inc.) as an NO-releasing macromolecule is incorporated in the foam at a loading of 1% (w/w).
There are two moles of NH per mole of 2-(2-aminoethylamino)ethanol. The number of moles of 2-(2-aminoethylamino) ethanol in 100 kg=961.54. The number of moles of NH=2*961.54=1923.07 moles. Thus, the weight of CO2 required=1923.07 moles*44 g/mol=84.6 kg. A 1.2 fold excess gas to ensure complete conversion to carbamates. Thus, the amount of CO2 required=101.52 kg.
Calculation for Catalyst Addition
Two types of catalyst are used in combination, a gel catalyst to accelerate the urethane formation reaction, and a blowing catalyst to reduce the rising time of the foam. The gel catalyst (e.g., stannous octoate) is present as 0.3 parts per 100 parts polyol (w/w). The blowing catalyst (e.g., antimony tris 2-ethylhexoate) is present as 0.3 parts per 100 parts polyol (w/w). Therefore, the catalyst calculation is based on the total mass of reacting hydroxyl compounds, and includes the hydroxyls in 2-(2-aminoethylamino) ethanol. Thus, the weight of gel catalyst=300 kg/100 kg* 0.3=9 kg. The weight of blowing catalyst=9 kg.
Calculation for Desmodur N-100 Addition
Enough Desmodur N-100 needs to be added such that the NCO are adequate to react with OH in 2-(2-aminoethylamino) ethanol and in Desmophen. The equivalent of OH groups in Desmophen-R-221-75 (3.3% OH)=17*100/3.3=515. The moles of OH groups in 100 kg 2-(2-aminoethylamino)ethanol=961.54. The equivalents in 200 kg=200/515=0.388. The equivalent NCO groups in Desmodur N-100 (22% NCO)=42*100/22=191. The equivalents of NCO required for 1:1 reaction with 200 kg Desmophen-R-221-75=191*0.388=74.2 kg. The percent OH in 2-(2-aminoethylamino)ethanol=17/104=16.3%. The equivalent of OH groups=17*100/16.3=104. The equivalents in 100 kg=100/104=0.9615. The equivalents of NCO required for 1:1 reaction=0.9615*191=183.65 kg. Thus, the total required=183.65 6+74.2=257.83 kg. A 2% excess is used to ensure complete reaction. Therefore, the Desmodur N-100 total requirement=1.02*257.83=262.985 kg.
Calculation for Nitricil™-70
The total reactant weight (not including CO2)=262.985 kg (Desmodur N-100)+200 kg (Desmophen-R-221-75 polyol)+100 kg (2-(2-aminoethylamino) ethanol)+18 kg (weight of blowing catalyst and gel catalyst)=580.985 kg. Thus, 1% Nitricil™-70=5.81 kg.
In the drawings and specification, there have been disclosed typical embodiments of the invention and, although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation, the scope of the invention being set forth in the following claims.
The present application is a 35 U.S.C. § 371 national phase application of PCT International Application No. PCT/US2010/046209, having an international filing date of Aug. 20, 2010, claiming priority to U.S. Provisional Application Serial No. 61/235,927, filed Aug. 21, 2009, and U.S. Provisional Application Serial No. 61/235,948, filed Aug. 21, 2009. The disclosures of each application are incorporated herein by reference in their entireties. The above PCT International Application was published in the English language and has been accorded International Publication No. WO 2011/022680 A2.
Research for this application was partially funded through a Phase I NIH SBIR grant entitled “Nitric Oxide-Releasing Antibacterial Wound Dressing” (grant number 5R43AI074098-02). The government may have certain rights to this application.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/US2010/046209 | 8/20/2010 | WO | 00 | 10/4/2011 |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2011/022680 | 2/24/2011 | WO | A |
| Number | Name | Date | Kind |
|---|---|---|---|
| 4182827 | Jones | Jan 1980 | A |
| 4507466 | Tomalia et al. | Mar 1985 | A |
| 4550126 | Lorenz | Oct 1985 | A |
| 4558120 | Tomalia et al. | Dec 1985 | A |
| 4568737 | Tomalia et al. | Feb 1986 | A |
| 4587329 | Tomalia et al. | May 1986 | A |
| 4600001 | Gilman | Jul 1986 | A |
| 4631337 | Tomalia et al. | Dec 1986 | A |
| 4694064 | Tomalia et al. | Sep 1987 | A |
| 4713975 | Tomalia et al. | Dec 1987 | A |
| 4737550 | Tomalia | Apr 1988 | A |
| 4857599 | Tomalia et al. | Aug 1989 | A |
| 4871779 | Killat et al. | Oct 1989 | A |
| 4985023 | Blank et al. | Jan 1991 | A |
| 4990338 | Blank et al. | Feb 1991 | A |
| 5035892 | Blank et al. | Jul 1991 | A |
| 5045322 | Blank et al. | Sep 1991 | A |
| 5061487 | Blank et al. | Oct 1991 | A |
| 5079004 | Blank et al. | Jan 1992 | A |
| 5380758 | Stamler et al. | Jan 1995 | A |
| 5405919 | Keefer et al. | Apr 1995 | A |
| 5418301 | Hult et al. | May 1995 | A |
| 5428070 | Cooke et al. | Jun 1995 | A |
| 5504117 | Gorfine | Apr 1996 | A |
| 5519020 | Smith et al. | May 1996 | A |
| 5525357 | Keefer et al. | Jun 1996 | A |
| 5574068 | Stamler et al. | Nov 1996 | A |
| 5593876 | Stamler et al. | Jan 1997 | A |
| 5599984 | Bianchi et al. | Feb 1997 | A |
| 5629322 | Guthikonda et al. | May 1997 | A |
| 5632981 | Saavedra et al. | May 1997 | A |
| 5650442 | Mitchell et al. | Jul 1997 | A |
| 5650447 | Keefer et al. | Jul 1997 | A |
| 5665077 | Rosen et al. | Sep 1997 | A |
| 5676963 | Keefer et al. | Oct 1997 | A |
| 5691423 | Smith et al. | Nov 1997 | A |
| 5693676 | Gorfine | Dec 1997 | A |
| 5700830 | Korthuis et al. | Dec 1997 | A |
| 5718892 | Keefer et al. | Feb 1998 | A |
| 5726156 | Girten et al. | Mar 1998 | A |
| 5750573 | Bianchi et al. | May 1998 | A |
| 5753684 | Bianchi et al. | May 1998 | A |
| 5760001 | Girten et al. | Jun 1998 | A |
| 5770645 | Stamler et al. | Jun 1998 | A |
| 5786332 | Girten et al. | Jul 1998 | A |
| 5789447 | Wink, Jr. et al. | Aug 1998 | A |
| 5797887 | Rosen et al. | Aug 1998 | A |
| 5810010 | Anbar | Sep 1998 | A |
| 5814666 | Green et al. | Sep 1998 | A |
| 5814667 | Mitchell et al. | Sep 1998 | A |
| 5821261 | Durette et al. | Oct 1998 | A |
| 5837736 | Mitchell et al. | Nov 1998 | A |
| 5840759 | Mitchell et al. | Nov 1998 | A |
| 5849794 | Bianchi et al. | Dec 1998 | A |
| 5852058 | Cooke et al. | Dec 1998 | A |
| 5854289 | Bianchi et al. | Dec 1998 | A |
| 5859062 | Bianchi et al. | Jan 1999 | A |
| 5861168 | Cooke et al. | Jan 1999 | A |
| 5863890 | Stamler et al. | Jan 1999 | A |
| 5891459 | Cooke et al. | Apr 1999 | A |
| 5891472 | Russell | Apr 1999 | A |
| 5910316 | Keefer et al. | Jun 1999 | A |
| 5914125 | Andrews | Jun 1999 | A |
| 5932538 | Garvey et al. | Aug 1999 | A |
| 5958427 | Salzman et al. | Sep 1999 | A |
| 5961466 | Anbar | Oct 1999 | A |
| 5962520 | Smith et al. | Oct 1999 | A |
| 5968528 | Deckner et al. | Oct 1999 | A |
| 5994294 | Garvey et al. | Nov 1999 | A |
| 5994444 | Trescony et al. | Nov 1999 | A |
| 5999843 | Anbar | Dec 1999 | A |
| 6008255 | Bianchi et al. | Dec 1999 | A |
| 6022900 | Bianchi et al. | Feb 2000 | A |
| 6035225 | Anbar | Mar 2000 | A |
| 6043358 | Caldwell et al. | Mar 2000 | A |
| 6045827 | Russell | Apr 2000 | A |
| 6070928 | Campbell | Jun 2000 | A |
| 6087479 | Stamler et al. | Jul 2000 | A |
| 6103266 | Tapolsky et al. | Aug 2000 | A |
| 6103275 | Seitz et al. | Aug 2000 | A |
| 6110453 | Keefer et al. | Aug 2000 | A |
| 6143037 | Goldstein et al. | Nov 2000 | A |
| 6147068 | Smith et al. | Nov 2000 | A |
| 6151522 | Alfano et al. | Nov 2000 | A |
| 6160021 | Lerner et al. | Dec 2000 | A |
| 6171232 | Papandreou et al. | Jan 2001 | B1 |
| 6174539 | Stamler et al. | Jan 2001 | B1 |
| 6180082 | Woltering et al. | Jan 2001 | B1 |
| 6180676 | Bianchi et al. | Jan 2001 | B1 |
| 6190704 | Murrell | Feb 2001 | B1 |
| 6200558 | Saavedra et al. | Mar 2001 | B1 |
| 6207855 | Toone et al. | Mar 2001 | B1 |
| 6218016 | Tedeschi et al. | Apr 2001 | B1 |
| 6232336 | Hrabie et al. | May 2001 | B1 |
| 6232434 | Stamler et al. | May 2001 | B1 |
| 6238683 | Burnett et al. | May 2001 | B1 |
| 6248787 | Bianchi et al. | Jun 2001 | B1 |
| 6255277 | Stamler et al. | Jul 2001 | B1 |
| 6261594 | Smith et al. | Jul 2001 | B1 |
| 6270779 | Fitzhugh et al. | Aug 2001 | B1 |
| 6287601 | Russell | Sep 2001 | B1 |
| 6290981 | Keefer et al. | Sep 2001 | B1 |
| 6291424 | Stamler et al. | Sep 2001 | B1 |
| 6294517 | Garvey et al. | Sep 2001 | B1 |
| 6299980 | Shah et al. | Oct 2001 | B1 |
| 6323211 | Garvey et al. | Nov 2001 | B1 |
| 6350467 | Demopoulos et al. | Feb 2002 | B1 |
| 6352709 | Stamler et al. | Mar 2002 | B1 |
| 6358536 | Thomas | Mar 2002 | B1 |
| 6359167 | Toone et al. | Mar 2002 | B2 |
| 6359182 | Stamler et al. | Mar 2002 | B1 |
| 6369071 | Haj-Yehia | Apr 2002 | B1 |
| 6372733 | Caldwell et al. | Apr 2002 | B1 |
| 6377321 | Khan et al. | Apr 2002 | B1 |
| 6379660 | Saavedra et al. | Apr 2002 | B1 |
| 6379691 | Tedeschi et al. | Apr 2002 | B1 |
| 6391895 | Towart et al. | May 2002 | B1 |
| 6403759 | Stamler et al. | Jun 2002 | B2 |
| 6410622 | Endres | Jun 2002 | B1 |
| 6417162 | Garvey et al. | Jul 2002 | B1 |
| 6432077 | Stenzler | Aug 2002 | B1 |
| 6433182 | Garvey et al. | Aug 2002 | B1 |
| 6436975 | Del Soldato | Aug 2002 | B1 |
| 6441254 | Dobert | Aug 2002 | B1 |
| 6448267 | Anggard et al. | Sep 2002 | B1 |
| 6451337 | Smith et al. | Sep 2002 | B1 |
| 6455542 | Anggard et al. | Sep 2002 | B1 |
| 6469065 | Garvey et al. | Oct 2002 | B1 |
| 6471978 | Stamler et al. | Oct 2002 | B1 |
| 6472390 | Stamler et al. | Oct 2002 | B1 |
| 6474508 | Marsh | Nov 2002 | B1 |
| 6488951 | Toone et al. | Dec 2002 | B2 |
| 6492405 | Haj-Yehia | Dec 2002 | B2 |
| 6511991 | Hrabie et al. | Jan 2003 | B2 |
| 6514934 | Garvey et al. | Feb 2003 | B1 |
| 6538033 | Bing | Mar 2003 | B2 |
| 6560478 | Alfano et al. | May 2003 | B1 |
| 6562344 | Stamler et al. | May 2003 | B1 |
| 6562785 | Shapiro | May 2003 | B1 |
| 6583113 | Stamler et al. | Jun 2003 | B2 |
| 6583311 | Toone et al. | Jun 2003 | B2 |
| 6605447 | Weiss et al. | Aug 2003 | B2 |
| 6610660 | Saavedra et al. | Aug 2003 | B1 |
| 6627602 | Stamler et al. | Sep 2003 | B2 |
| 6642208 | Cooke et al. | Nov 2003 | B2 |
| 6642260 | Haj-Yehia | Nov 2003 | B2 |
| 6645518 | Tedeschi et al. | Nov 2003 | B2 |
| 6646006 | Cooke et al. | Nov 2003 | B2 |
| 6656217 | Herzog, Jr. et al. | Dec 2003 | B1 |
| 6673338 | Arnold et al. | Jan 2004 | B1 |
| 6673891 | Stamler et al. | Jan 2004 | B2 |
| 6699846 | Elliott et al. | Mar 2004 | B2 |
| 6703046 | Fitzhugh et al. | Mar 2004 | B2 |
| 6706274 | Hermann et al. | Mar 2004 | B2 |
| 6709681 | Benjamin et al. | Mar 2004 | B2 |
| 6723703 | Gaston et al. | Apr 2004 | B2 |
| 6737447 | Smith et al. | May 2004 | B1 |
| 6747062 | Murrell | Jun 2004 | B2 |
| 6750254 | Hrabie et al. | Jun 2004 | B2 |
| 6758214 | Fine et al. | Jul 2004 | B2 |
| 6759430 | Anggard et al. | Jul 2004 | B2 |
| 6780849 | Herrmann et al. | Aug 2004 | B2 |
| 6793644 | Stenzler | Sep 2004 | B2 |
| 6796966 | Thomas | Sep 2004 | B2 |
| 6841166 | Zhang et al. | Jan 2005 | B1 |
| 6855366 | Smith et al. | Feb 2005 | B2 |
| 6875840 | Stamler et al. | Apr 2005 | B2 |
| 6887485 | Fitzhugh et al. | May 2005 | B2 |
| 6887994 | Stamler et al. | May 2005 | B2 |
| 6894073 | Lee et al. | May 2005 | B2 |
| 6896899 | Demopolos et al. | May 2005 | B2 |
| 6897218 | Casella et al. | May 2005 | B2 |
| 6911433 | Saavedra et al. | Jun 2005 | B2 |
| 6911478 | Hrabie et al. | Jun 2005 | B2 |
| 6946484 | Adams et al. | Sep 2005 | B2 |
| 6949530 | Hrabie et al. | Sep 2005 | B2 |
| 6951902 | McDonald et al. | Oct 2005 | B2 |
| 6964984 | Stamler et al. | Nov 2005 | B2 |
| 6974801 | Honda et al. | Dec 2005 | B2 |
| 7012098 | Manning et al. | Mar 2006 | B2 |
| 7015347 | Toone et al. | Mar 2006 | B2 |
| 7025869 | Fine et al. | Apr 2006 | B2 |
| 7030238 | Stamler et al. | Apr 2006 | B2 |
| 7033999 | Stamler et al. | Apr 2006 | B2 |
| 7040313 | Fine et al. | May 2006 | B2 |
| 7048951 | Seitz et al. | May 2006 | B1 |
| 7049308 | Stamler et al. | May 2006 | B2 |
| 7052711 | West et al. | May 2006 | B2 |
| 7070798 | Michal et al. | Jul 2006 | B1 |
| 7081524 | Saavedra et al. | Jul 2006 | B2 |
| 7087588 | Del Soldato | Aug 2006 | B2 |
| 7087709 | Stamler et al. | Aug 2006 | B2 |
| 7122018 | Stenzler et al. | Oct 2006 | B2 |
| 7122027 | Trescony et al. | Oct 2006 | B2 |
| 7122529 | Ruane et al. | Oct 2006 | B2 |
| 7128904 | Batchelor et al. | Oct 2006 | B2 |
| 7135189 | Knapp | Nov 2006 | B2 |
| 7135498 | Chopp et al. | Nov 2006 | B1 |
| 7157500 | Stamler et al. | Jan 2007 | B2 |
| 7169809 | Berthelette et al. | Jan 2007 | B2 |
| 7176237 | Honda et al. | Feb 2007 | B2 |
| 7179475 | Burnett et al. | Feb 2007 | B1 |
| 7189761 | Gorfine | Mar 2007 | B2 |
| 7199154 | Berthelette et al. | Apr 2007 | B2 |
| 7204980 | Clark | Apr 2007 | B2 |
| 7226586 | Fitzhugh et al. | Jun 2007 | B2 |
| 7234079 | Cheng | Jun 2007 | B2 |
| 7259250 | Stamler et al. | Aug 2007 | B2 |
| 7279176 | West et al. | Oct 2007 | B1 |
| 7282519 | Garvey et al. | Oct 2007 | B2 |
| 7303575 | Ogle | Dec 2007 | B2 |
| 7314857 | Madhyastha | Jan 2008 | B2 |
| 7335383 | Meyerhoff et al. | Feb 2008 | B2 |
| 7345053 | Garvey | Mar 2008 | B2 |
| 7348319 | Hrabie et al. | Mar 2008 | B2 |
| 7394585 | Weber | Apr 2008 | B2 |
| 7396829 | Garvey et al. | Jul 2008 | B2 |
| 7417109 | Stamler et al. | Aug 2008 | B2 |
| 7425218 | Keefer et al. | Sep 2008 | B2 |
| 7432301 | Gaston et al. | Oct 2008 | B2 |
| 7452916 | Cooke | Nov 2008 | B2 |
| 7468435 | Waterhouse et al. | Dec 2008 | B2 |
| 7485324 | Miller et al. | Feb 2009 | B2 |
| 7520866 | Stenzler et al. | Apr 2009 | B2 |
| 7531164 | Daaka et al. | May 2009 | B2 |
| 7569559 | Arnold et al. | Aug 2009 | B2 |
| 7582623 | Mascharak | Sep 2009 | B2 |
| 7595313 | Garvey et al. | Sep 2009 | B2 |
| 7622501 | Dufresne et al. | Nov 2009 | B2 |
| 7622502 | Berthelette et al. | Nov 2009 | B2 |
| 7645748 | Orchansky et al. | Jan 2010 | B2 |
| 7645749 | Orchansky et al. | Jan 2010 | B2 |
| 7651697 | West et al. | Jan 2010 | B2 |
| 7655423 | Chopp et al. | Feb 2010 | B2 |
| 7678391 | Graham et al. | Mar 2010 | B2 |
| 7678830 | Honda et al. | Mar 2010 | B2 |
| 7696247 | Herrmann et al. | Apr 2010 | B2 |
| 7704518 | Tamarkin et al. | Apr 2010 | B2 |
| 7745656 | Toone et al. | Jun 2010 | B2 |
| 7763283 | Batchelor et al. | Jul 2010 | B2 |
| 7785616 | Stamler et al. | Aug 2010 | B2 |
| 7795286 | Lucet-Levannier | Sep 2010 | B2 |
| 7799335 | Herrmann et al. | Sep 2010 | B2 |
| 7807716 | Farber | Oct 2010 | B2 |
| 7811600 | Cheng et al. | Oct 2010 | B2 |
| 7820284 | Terry | Oct 2010 | B2 |
| 7829553 | Arnold et al. | Nov 2010 | B2 |
| 7838023 | Garvey et al. | Nov 2010 | B2 |
| 7846400 | Hyde et al. | Dec 2010 | B2 |
| 7862598 | Hyde et al. | Jan 2011 | B2 |
| 7892198 | Stenzler | Feb 2011 | B2 |
| 7897399 | Hyde et al. | Mar 2011 | B2 |
| 7928079 | Hrabie et al. | Apr 2011 | B2 |
| 7928096 | Waterhouse et al. | Apr 2011 | B2 |
| 7947299 | Knapp | May 2011 | B2 |
| 7972137 | Rosen | Jul 2011 | B2 |
| 7975699 | Hyde et al. | Jul 2011 | B2 |
| 8003811 | Almirante | Aug 2011 | B2 |
| 8017074 | Arnold | Sep 2011 | B2 |
| 8021679 | Chen | Sep 2011 | B2 |
| 8034384 | Meyerhoff | Oct 2011 | B2 |
| 8043246 | Av-Gay et al. | Oct 2011 | B2 |
| 8241650 | Peters | Aug 2012 | B2 |
| 8343945 | Tamarkin et al. | Jan 2013 | B2 |
| 8362091 | Tamarkin et al. | Jan 2013 | B2 |
| 8399005 | Schoenfisch | Mar 2013 | B2 |
| 8486374 | Tamarkin et al. | Jul 2013 | B2 |
| 8617100 | Eini et al. | Dec 2013 | B2 |
| 8636982 | Tamarkin et al. | Jan 2014 | B2 |
| 8795635 | Tamarkin et al. | Aug 2014 | B2 |
| 8795693 | Tamarkin et al. | Aug 2014 | B2 |
| 8900553 | Tamarkin et al. | Dec 2014 | B2 |
| 9101662 | Tamarkin et al. | Aug 2015 | B2 |
| 9161916 | Tamarkin et al. | Oct 2015 | B2 |
| 9265725 | Tamarkin et al. | Feb 2016 | B2 |
| 9320705 | Tamarkin et al. | Apr 2016 | B2 |
| 20010012851 | Lundy et al. | Aug 2001 | A1 |
| 20010025057 | Gorfine | Sep 2001 | A1 |
| 20010038832 | Bonavida et al. | Nov 2001 | A1 |
| 20010053772 | Bonavida et al. | Dec 2001 | A1 |
| 20020013304 | Wilson et al. | Jan 2002 | A1 |
| 20020018757 | Harichian et al. | Feb 2002 | A1 |
| 20020028223 | Vatter et al. | Mar 2002 | A1 |
| 20020028851 | Bianchi et al. | Mar 2002 | A1 |
| 20020049157 | Wu et al. | Apr 2002 | A1 |
| 20020061879 | Garvey et al. | May 2002 | A1 |
| 20020068365 | Kuhrts | Jun 2002 | A1 |
| 20020072550 | Brady | Jun 2002 | A1 |
| 20020090401 | Tucker et al. | Jul 2002 | A1 |
| 20020094985 | Herrmann | Jul 2002 | A1 |
| 20020115586 | Enikolopov | Aug 2002 | A1 |
| 20020122929 | Simpson et al. | Sep 2002 | A1 |
| 20020132234 | Moskowitz | Sep 2002 | A1 |
| 20020133040 | Woo et al. | Sep 2002 | A1 |
| 20020136763 | Demopoulos et al. | Sep 2002 | A1 |
| 20020138051 | Hole et al. | Sep 2002 | A1 |
| 20020143007 | Garvey et al. | Oct 2002 | A1 |
| 20020143062 | Lopez-Berestein et al. | Oct 2002 | A1 |
| 20020155174 | Benjamin et al. | Oct 2002 | A1 |
| 20020161042 | Gorfine | Oct 2002 | A1 |
| 20030027844 | Soldato | Feb 2003 | A1 |
| 20030039697 | Zhao et al. | Feb 2003 | A1 |
| 20030050305 | Tejada | Mar 2003 | A1 |
| 20030072783 | Stamler et al. | Apr 2003 | A1 |
| 20030093143 | Zhao et al. | May 2003 | A1 |
| 20030134779 | Diarra et al. | Jul 2003 | A1 |
| 20030170674 | Moskowitz | Sep 2003 | A1 |
| 20030203915 | Fang et al. | Oct 2003 | A1 |
| 20030219854 | Guarna et al. | Nov 2003 | A1 |
| 20040009238 | Miller et al. | Jan 2004 | A1 |
| 20040013747 | Tucker et al. | Jan 2004 | A1 |
| 20040033480 | Wong | Feb 2004 | A1 |
| 20040037836 | Stamler et al. | Feb 2004 | A1 |
| 20040037897 | Benjamin et al. | Feb 2004 | A1 |
| 20040043068 | Tedeschi et al. | Mar 2004 | A1 |
| 20040076582 | Dimatteo et al. | Apr 2004 | A1 |
| 20040082659 | Cooke et al. | Apr 2004 | A1 |
| 20040105898 | Benjamin et al. | Jun 2004 | A1 |
| 20040110691 | Stamler | Jun 2004 | A1 |
| 20040131703 | Bach et al. | Jul 2004 | A1 |
| 20040143010 | Esteve-Soler et al. | Jul 2004 | A1 |
| 20040147598 | Haj-Yehia | Jul 2004 | A1 |
| 20040157936 | Burnett et al. | Aug 2004 | A1 |
| 20040228889 | Cals-Grierson | Nov 2004 | A1 |
| 20040254419 | Wang et al. | Dec 2004 | A1 |
| 20040265244 | Rosen | Dec 2004 | A1 |
| 20050036949 | Tucker et al. | Feb 2005 | A1 |
| 20050037093 | Benjamin | Feb 2005 | A1 |
| 20050038473 | Tamarkin et al. | Feb 2005 | A1 |
| 20050054714 | Munoz et al. | Mar 2005 | A1 |
| 20050065161 | Garvey et al. | Mar 2005 | A1 |
| 20050069595 | Chen et al. | Mar 2005 | A1 |
| 20050074506 | Natan et al. | Apr 2005 | A1 |
| 20050079132 | Wang et al. | Apr 2005 | A1 |
| 20050080021 | Tucker et al. | Apr 2005 | A1 |
| 20050080024 | Tucker et al. | Apr 2005 | A1 |
| 20050131064 | Gaston et al. | Jun 2005 | A1 |
| 20050142217 | Adams et al. | Jun 2005 | A1 |
| 20050142218 | Tucker et al. | Jun 2005 | A1 |
| 20050152891 | Toone et al. | Jul 2005 | A1 |
| 20050165452 | Sigg et al. | Jul 2005 | A1 |
| 20050171066 | Bunting et al. | Aug 2005 | A1 |
| 20050171199 | Murrell | Aug 2005 | A1 |
| 20050187222 | Garvey et al. | Aug 2005 | A1 |
| 20050220838 | Zhao et al. | Oct 2005 | A1 |
| 20050232869 | Tamarkin et al. | Oct 2005 | A1 |
| 20050245492 | Lephart et al. | Nov 2005 | A1 |
| 20050249818 | Sawan et al. | Nov 2005 | A1 |
| 20050265958 | West et al. | Dec 2005 | A1 |
| 20050271596 | Friedman et al. | Dec 2005 | A1 |
| 20050281867 | Kahn et al. | Dec 2005 | A1 |
| 20060008529 | Meyerhoff et al. | Jan 2006 | A1 |
| 20060009431 | Earl et al. | Jan 2006 | A1 |
| 20060035854 | Goldstein et al. | Feb 2006 | A1 |
| 20060039950 | Zhou et al. | Feb 2006 | A1 |
| 20060058363 | Wang et al. | Mar 2006 | A1 |
| 20060067909 | West et al. | Mar 2006 | A1 |
| 20060095120 | Hermann | May 2006 | A1 |
| 20060100159 | Stamler et al. | May 2006 | A1 |
| 20060142183 | Diarra et al. | Jun 2006 | A1 |
| 20060147553 | Miller et al. | Jul 2006 | A1 |
| 20060147904 | Wong | Jul 2006 | A1 |
| 20060153904 | Smith et al. | Jul 2006 | A1 |
| 20060155260 | Blott et al. | Jul 2006 | A1 |
| 20060159726 | Shell | Jul 2006 | A1 |
| 20060172018 | Fine et al. | Aug 2006 | A1 |
| 20060198831 | Stamler et al. | Sep 2006 | A1 |
| 20060211601 | Stamler et al. | Sep 2006 | A1 |
| 20060240065 | Chen | Oct 2006 | A1 |
| 20060269620 | Morris et al. | Nov 2006 | A1 |
| 20060275218 | Tamarkin et al. | Dec 2006 | A1 |
| 20060286158 | Calvert Murrell et al. | Dec 2006 | A1 |
| 20060286159 | Calvert Murrell et al. | Dec 2006 | A1 |
| 20070003538 | Madhyastha | Jan 2007 | A1 |
| 20070014686 | Arnold et al. | Jan 2007 | A1 |
| 20070014733 | O'Donnell et al. | Jan 2007 | A1 |
| 20070014828 | Fitzhugh et al. | Jan 2007 | A1 |
| 20070037821 | Garvey et al. | Feb 2007 | A1 |
| 20070048344 | Yahiaoui et al. | Mar 2007 | A1 |
| 20070053952 | Chen et al. | Mar 2007 | A1 |
| 20070053955 | Larson et al. | Mar 2007 | A1 |
| 20070053966 | Ang et al. | Mar 2007 | A1 |
| 20070059338 | Knapp | Mar 2007 | A1 |
| 20070059351 | Murrell et al. | Mar 2007 | A1 |
| 20070086954 | Miller | Apr 2007 | A1 |
| 20070087025 | Fitzhugh et al. | Apr 2007 | A1 |
| 20070088345 | Larson et al. | Apr 2007 | A1 |
| 20070089739 | Fine et al. | Apr 2007 | A1 |
| 20070116785 | Miller | May 2007 | A1 |
| 20070129690 | Rosenblatt et al. | Jun 2007 | A1 |
| 20070148136 | Whitlock | Jun 2007 | A1 |
| 20070154570 | Miller et al. | Jul 2007 | A1 |
| 20070166227 | Liu et al. | Jul 2007 | A1 |
| 20070166255 | Gupta | Jul 2007 | A1 |
| 20070172469 | Clark | Jul 2007 | A1 |
| 20070191377 | Worcel | Aug 2007 | A1 |
| 20070196327 | Kalivretenos et al. | Aug 2007 | A1 |
| 20070197543 | Esteve-Soler et al. | Aug 2007 | A1 |
| 20070202155 | Ang et al. | Aug 2007 | A1 |
| 20070203242 | Calton | Aug 2007 | A1 |
| 20070207179 | Andersen et al. | Sep 2007 | A1 |
| 20070219208 | Kalyanaraman et al. | Sep 2007 | A1 |
| 20070225250 | Brown | Sep 2007 | A1 |
| 20070239107 | Lundberg et al. | Oct 2007 | A1 |
| 20070243262 | Hurley et al. | Oct 2007 | A1 |
| 20070248676 | Stamler et al. | Oct 2007 | A1 |
| 20070264225 | Cheng et al. | Nov 2007 | A1 |
| 20070270348 | Kahn et al. | Nov 2007 | A1 |
| 20070275100 | Miller | Nov 2007 | A1 |
| 20070292359 | Friedman et al. | Dec 2007 | A1 |
| 20070292461 | Tamarkin et al. | Dec 2007 | A1 |
| 20080025972 | Daaka et al. | Jan 2008 | A1 |
| 20080031907 | Tamarkin et al. | Feb 2008 | A1 |
| 20080033334 | Gurtner et al. | Feb 2008 | A1 |
| 20080039521 | Yasuda et al. | Feb 2008 | A1 |
| 20080044444 | Tamarkin et al. | Feb 2008 | A1 |
| 20080045909 | Fossel | Feb 2008 | A1 |
| 20080063607 | Tamarkin et al. | Mar 2008 | A1 |
| 20080069779 | Tamarkin et al. | Mar 2008 | A1 |
| 20080069848 | Peters | Mar 2008 | A1 |
| 20080069863 | Peters | Mar 2008 | A1 |
| 20080069905 | Peters | Mar 2008 | A1 |
| 20080071206 | Peters | Mar 2008 | A1 |
| 20080089956 | Da et al. | Apr 2008 | A1 |
| 20080138296 | Tamarkin et al. | Jun 2008 | A1 |
| 20080139450 | Madhyastha et al. | Jun 2008 | A1 |
| 20080145449 | Stamler | Jun 2008 | A1 |
| 20080152596 | Friedman et al. | Jun 2008 | A1 |
| 20080166303 | Tamarkin et al. | Jul 2008 | A1 |
| 20080171021 | Bach et al. | Jul 2008 | A1 |
| 20080171351 | Smith | Jul 2008 | A1 |
| 20080175881 | Ippoliti et al. | Jul 2008 | A1 |
| 20080182797 | Nudler et al. | Jul 2008 | A1 |
| 20080193385 | Maibach | Aug 2008 | A1 |
| 20080193566 | Miller et al. | Aug 2008 | A1 |
| 20080206155 | Tamarkin et al. | Aug 2008 | A1 |
| 20080206159 | Tamarkin et al. | Aug 2008 | A1 |
| 20080206161 | Tamarkin et al. | Aug 2008 | A1 |
| 20080207491 | Diarra et al. | Aug 2008 | A1 |
| 20080207713 | Wang et al. | Aug 2008 | A1 |
| 20080214646 | Knaus et al. | Sep 2008 | A1 |
| 20080226751 | Tucker et al. | Sep 2008 | A1 |
| 20080241208 | Shanley et al. | Oct 2008 | A1 |
| 20080253973 | Tamarkin et al. | Oct 2008 | A1 |
| 20080260655 | Tamarkin et al. | Oct 2008 | A1 |
| 20080275093 | Garvey et al. | Nov 2008 | A1 |
| 20080280984 | Fossel | Nov 2008 | A1 |
| 20080286321 | Reneker et al. | Nov 2008 | A1 |
| 20080287861 | Stenzler et al. | Nov 2008 | A1 |
| 20080292560 | Tamarkin et al. | Nov 2008 | A1 |
| 20080299220 | Tamarkin et al. | Dec 2008 | A1 |
| 20080306012 | Hrabie et al. | Dec 2008 | A1 |
| 20080311163 | Peters | Dec 2008 | A1 |
| 20080317626 | Arnold et al. | Dec 2008 | A1 |
| 20080317679 | Tamarkin et al. | Dec 2008 | A1 |
| 20090004298 | Gaston et al. | Jan 2009 | A1 |
| 20090010989 | Peters | Jan 2009 | A1 |
| 20090018091 | Ellis et al. | Jan 2009 | A1 |
| 20090028966 | Chen et al. | Jan 2009 | A1 |
| 20090029028 | Garcin et al. | Jan 2009 | A1 |
| 20090036491 | Tucker et al. | Feb 2009 | A1 |
| 20090041680 | Tamarkin et al. | Feb 2009 | A1 |
| 20090042819 | Ellis et al. | Feb 2009 | A1 |
| 20090048219 | Garvey | Feb 2009 | A1 |
| 20090068118 | Eini et al. | Mar 2009 | A1 |
| 20090069449 | Smith et al. | Mar 2009 | A1 |
| 20090081279 | Jezek et al. | Mar 2009 | A1 |
| 20090088411 | Renzi et al. | Apr 2009 | A1 |
| 20090093510 | Clementi et al. | Apr 2009 | A1 |
| 20090098187 | Peters et al. | Apr 2009 | A1 |
| 20090108777 | Hyde et al. | Apr 2009 | A1 |
| 20090110612 | Hyde et al. | Apr 2009 | A1 |
| 20090110712 | Hyde et al. | Apr 2009 | A1 |
| 20090110933 | Hyde et al. | Apr 2009 | A1 |
| 20090110958 | Hyde et al. | Apr 2009 | A1 |
| 20090112055 | Hyde et al. | Apr 2009 | A1 |
| 20090112193 | Hyde et al. | Apr 2009 | A1 |
| 20090112197 | Hyde et al. | Apr 2009 | A1 |
| 20090118819 | Merz et al. | May 2009 | A1 |
| 20090123528 | Fossel | May 2009 | A1 |
| 20090130029 | Tamarkin et al. | May 2009 | A1 |
| 20090131342 | Ellis | May 2009 | A1 |
| 20090136410 | Smith | May 2009 | A1 |
| 20090137683 | Yasuda et al. | May 2009 | A1 |
| 20090143417 | Smith et al. | Jun 2009 | A1 |
| 20090148482 | Peters | Jun 2009 | A1 |
| 20090175799 | Tamarkin et al. | Jul 2009 | A1 |
| 20090177133 | Kieswetter | Jul 2009 | A1 |
| 20090186859 | Velázquez et al. | Jul 2009 | A1 |
| 20090191284 | Conoci et al. | Jul 2009 | A1 |
| 20090196930 | Surber et al. | Aug 2009 | A1 |
| 20090197964 | Summar et al. | Aug 2009 | A1 |
| 20090203653 | Garvey | Aug 2009 | A1 |
| 20090214618 | Schoenfisch et al. | Aug 2009 | A1 |
| 20090214624 | Smith et al. | Aug 2009 | A1 |
| 20090214674 | Barraud et al. | Aug 2009 | A1 |
| 20090215838 | Garvey et al. | Aug 2009 | A1 |
| 20090221536 | Fossel | Sep 2009 | A1 |
| 20090222088 | Chen et al. | Sep 2009 | A1 |
| 20090226504 | Peters | Sep 2009 | A1 |
| 20090232863 | Cheng et al. | Sep 2009 | A1 |
| 20090232868 | Chen et al. | Sep 2009 | A1 |
| 20090255536 | Av-Gay et al. | Oct 2009 | A1 |
| 20090263416 | Dawson et al. | Oct 2009 | A1 |
| 20090264398 | Bauer | Oct 2009 | A1 |
| 20090270509 | Arnold et al. | Oct 2009 | A1 |
| 20090287072 | Meyerhoff et al. | Nov 2009 | A1 |
| 20090297634 | Friedman et al. | Dec 2009 | A1 |
| 20090304815 | Cossu et al. | Dec 2009 | A1 |
| 20090317885 | Mascharak | Dec 2009 | A1 |
| 20100003338 | Hubbell et al. | Jan 2010 | A1 |
| 20100015253 | Benjamin | Jan 2010 | A1 |
| 20100016767 | Jones et al. | Jan 2010 | A1 |
| 20100016790 | Peters | Jan 2010 | A1 |
| 20100021506 | Jones | Jan 2010 | A1 |
| 20100040703 | Miller et al. | Feb 2010 | A1 |
| 20100062055 | Herrmann et al. | Mar 2010 | A1 |
| 20100076162 | Ameer et al. | Mar 2010 | A1 |
| 20100086530 | Martinov | Apr 2010 | A1 |
| 20100087370 | Jain et al. | Apr 2010 | A1 |
| 20100098733 | Stasko | Apr 2010 | A1 |
| 20100099729 | Almirante et al. | Apr 2010 | A1 |
| 20100112033 | Ganzarolli de Oliveira et al. | May 2010 | A1 |
| 20100112095 | Morris et al. | May 2010 | A1 |
| 20100129474 | Benjamin et al. | May 2010 | A1 |
| 20100152683 | Lindgren et al. | Jun 2010 | A1 |
| 20100159119 | Chen et al. | Jun 2010 | A1 |
| 20100166603 | Opie | Jul 2010 | A1 |
| 20100178319 | Lindgren et al. | Jul 2010 | A1 |
| 20100184992 | Toone et al. | Jul 2010 | A1 |
| 20100196517 | Fossel | Aug 2010 | A1 |
| 20100197702 | Hellberg et al. | Aug 2010 | A1 |
| 20100197802 | Jezek et al. | Aug 2010 | A1 |
| 20100209469 | Bezwada | Aug 2010 | A1 |
| 20100210745 | McDaniel et al. | Aug 2010 | A1 |
| 20100221195 | Tamarkin et al. | Sep 2010 | A1 |
| 20100221308 | Madhyastha et al. | Sep 2010 | A1 |
| 20100233304 | Pan | Sep 2010 | A1 |
| 20100239512 | Morris et al. | Sep 2010 | A1 |
| 20100247611 | Balkus, Jr. et al. | Sep 2010 | A1 |
| 20100247680 | Szabo | Sep 2010 | A1 |
| 20100255062 | Kalivretenos et al. | Oct 2010 | A1 |
| 20100256755 | Chen et al. | Oct 2010 | A1 |
| 20100256777 | Datta | Oct 2010 | A1 |
| 20100261930 | Honda et al. | Oct 2010 | A1 |
| 20100262238 | Chen et al. | Oct 2010 | A1 |
| 20100266510 | Tamarkin et al. | Oct 2010 | A1 |
| 20100268149 | Av-Gay et al. | Oct 2010 | A1 |
| 20100276284 | Meyerhoff et al. | Nov 2010 | A1 |
| 20100280122 | Fossel | Nov 2010 | A1 |
| 20100285100 | Balkus, Jr. et al. | Nov 2010 | A1 |
| 20100297200 | Schoenfisch et al. | Nov 2010 | A1 |
| 20100303891 | Lee et al. | Dec 2010 | A1 |
| 20100310476 | Tamarkin et al. | Dec 2010 | A1 |
| 20100311780 | Farber | Dec 2010 | A1 |
| 20100323036 | Fine | Dec 2010 | A1 |
| 20100324107 | Dos Santos et al. | Dec 2010 | A1 |
| 20100331542 | Smith | Dec 2010 | A1 |
| 20100331968 | Morris et al. | Dec 2010 | A1 |
| 20110008815 | Stamler et al. | Jan 2011 | A1 |
| 20110033437 | Smith et al. | Feb 2011 | A1 |
| 20110038965 | McKay et al. | Feb 2011 | A1 |
| 20110045037 | Tamarkin et al. | Feb 2011 | A1 |
| 20110046182 | Gilmer et al. | Feb 2011 | A1 |
| 20110059036 | Arnold et al. | Mar 2011 | A1 |
| 20110059189 | Cisneros | Mar 2011 | A1 |
| 20110065783 | O'Donnell et al. | Mar 2011 | A1 |
| 20110070318 | Jezek et al. | Mar 2011 | A1 |
| 20110071168 | Chopp et al. | Mar 2011 | A1 |
| 20110076313 | Av-Gay et al. | Mar 2011 | A1 |
| 20110086234 | Stasko et al. | Apr 2011 | A1 |
| 20110097279 | Tamarkin et al. | Apr 2011 | A1 |
| 20110104240 | Jones et al. | May 2011 | A1 |
| 20110106000 | Jones et al. | May 2011 | A1 |
| 20110212033 | Tamarkin et al. | Sep 2011 | A1 |
| 20120003293 | Miller | Jan 2012 | A1 |
| 20120141384 | Tamarkin et al. | Jun 2012 | A1 |
| 20120237453 | Tamarkin et al. | Sep 2012 | A1 |
| 20130164225 | Tamarkin et al. | Jun 2013 | A1 |
| 20130189191 | Tamarkin et al. | Jul 2013 | A1 |
| 20130189193 | Tamarkin et al. | Jul 2013 | A1 |
| 20130189195 | Tamarkin et al. | Jul 2013 | A1 |
| 20130196951 | Schoenfisch | Aug 2013 | A1 |
| 20140193502 | Tamarkin et al. | Jul 2014 | A1 |
| 20140248219 | Tamarkin et al. | Sep 2014 | A1 |
| 20140271494 | Tamarkin et al. | Sep 2014 | A1 |
| 20140369949 | Peters | Dec 2014 | A1 |
| 20150017103 | Tamarkin et al. | Jan 2015 | A1 |
| 20150025060 | Tamarkin et al. | Jan 2015 | A1 |
| 20150118164 | Tamarkin et al. | Apr 2015 | A1 |
| Number | Date | Country |
|---|---|---|
| 1387542 | Dec 2002 | CN |
| 1612804 | May 2005 | CN |
| 1819848 | Aug 2006 | CN |
| 101189032 | May 2008 | CN |
| 101242815 | Aug 2008 | CN |
| 101287505 | Oct 2008 | CN |
| 0 805 678 | Oct 2003 | EP |
| 0 746 327 | Apr 2004 | EP |
| 0 724 436 | Jul 2004 | EP |
| 1 411 908 | May 2005 | EP |
| 1 163 528 | Nov 2005 | EP |
| 1 681 068 | Jul 2006 | EP |
| 1 690 532 | Aug 2006 | EP |
| 1 690 554 | Aug 2006 | EP |
| 1 690 557 | Aug 2006 | EP |
| 1 690 558 | Aug 2006 | EP |
| 1 700 611 | Sep 2006 | EP |
| 1 704 876 | Sep 2006 | EP |
| 1 704 877 | Sep 2006 | EP |
| 1 704 879 | Sep 2006 | EP |
| 1 707 224 | Oct 2006 | EP |
| 1 728 438 | Dec 2006 | EP |
| 1 731 176 | Dec 2006 | EP |
| 1 757 278 | Feb 2007 | EP |
| 1 764 119 | Mar 2007 | EP |
| 1 790 335 | May 2007 | EP |
| 1 790 335 | May 2007 | EP |
| 1 861 130 | Sep 2008 | EP |
| 1 343 547 | Apr 2009 | EP |
| 1 871 433 | Apr 2009 | EP |
| 1 161 248 | May 2009 | EP |
| 1 846 058 | Jul 2009 | EP |
| 2 119 459 | Nov 2009 | EP |
| 2 142 179 | Jan 2010 | EP |
| 2 142 181 | Jan 2010 | EP |
| 1 917 005 | Sep 2010 | EP |
| WO 199507691 | Mar 1995 | WO |
| WO 199510267 | Apr 1995 | WO |
| WO 199512394 | May 1995 | WO |
| WO 199519767 | Jul 1995 | WO |
| WO 199522335 | Aug 1995 | WO |
| WO 199532715 | Dec 1995 | WO |
| WO 199608966 | Mar 1996 | WO |
| WO 199613164 | May 1996 | WO |
| WO 199614844 | May 1996 | WO |
| WO 1996015781 | May 1996 | WO |
| WO 199615797 | May 1996 | WO |
| WO 199627386 | Sep 1996 | WO |
| WO 199632118 | Oct 1996 | WO |
| WO 199632136 | Oct 1996 | WO |
| WO 1996033757 | Oct 1996 | WO |
| WO 199635416 | Nov 1996 | WO |
| WO 199716983 | May 1997 | WO |
| WO 199731654 | Sep 1997 | WO |
| WO 199734014 | Sep 1997 | WO |
| WO 1997047254 | Dec 1997 | WO |
| WO 1998005689 | Feb 1998 | WO |
| WO 199806389 | Feb 1998 | WO |
| WO 199808482 | Mar 1998 | WO |
| WO 199808482 | Mar 1998 | WO |
| WO 199808496 | Mar 1998 | WO |
| WO 199813358 | Apr 1998 | WO |
| WO 199819996 | May 1998 | WO |
| WO 1998020015 | May 1998 | WO |
| WO 199822090 | May 1998 | WO |
| WO 199829101 | Jul 1998 | WO |
| WO 199842661 | Oct 1998 | WO |
| WO 199900070 | Jan 1999 | WO |
| WO 199901427 | Jan 1999 | WO |
| WO 199918949 | Apr 1999 | WO |
| WO 199922729 | May 1999 | WO |
| WO 199933823 | Jul 1999 | WO |
| WO 199937616 | Jul 1999 | WO |
| WO 199944595 | Sep 1999 | WO |
| WO 199944595 | Sep 1999 | WO |
| WO 199951221 | Oct 1999 | WO |
| WO 199967210 | Dec 1999 | WO |
| WO 199967296 | Dec 1999 | WO |
| WO 200003640 | Jan 2000 | WO |
| WO 200006151 | Feb 2000 | WO |
| WO 200030658 | Jun 2000 | WO |
| WO 200033877 | Jun 2000 | WO |
| WO 200056333 | Sep 2000 | WO |
| WO 200059304 | Oct 2000 | WO |
| WO 200076318 | Dec 2000 | WO |
| WO 200112067 | Feb 2001 | WO |
| WO 2001015738 | Mar 2001 | WO |
| WO 2001015738 | Mar 2001 | WO |
| WO 200126702 | Apr 2001 | WO |
| WO 200126702 | Apr 2001 | WO |
| WO 200145732 | Jun 2001 | WO |
| WO 200145732 | Jun 2001 | WO |
| WO 200170199 | Sep 2001 | WO |
| WO 2001085227 | Nov 2001 | WO |
| WO 2001085227 | Nov 2001 | WO |
| WO 2001089572 | Nov 2001 | WO |
| WO 2002017880 | Mar 2002 | WO |
| WO 2002017880 | Mar 2002 | WO |
| WO 2002017881 | Mar 2002 | WO |
| WO 2002017881 | Mar 2002 | WO |
| WO 2002020026 | Mar 2002 | WO |
| WO 2002020026 | Mar 2002 | WO |
| WO 2002032418 | Apr 2002 | WO |
| WO 2002034705 | May 2002 | WO |
| WO 2002043786 | Jun 2002 | WO |
| WO 2002043786 | Jun 2002 | WO |
| WO 2002047675 | Jun 2002 | WO |
| WO 2002051353 | Jul 2002 | WO |
| WO 2002051353 | Jul 2002 | WO |
| WO 2002056864 | Jul 2002 | WO |
| WO 2002056864 | Jul 2002 | WO |
| WO 2002056874 | Jul 2002 | WO |
| WO 2002056904 | Jul 2002 | WO |
| WO 2002070496 | Sep 2002 | WO |
| WO 2002076395 | Oct 2002 | WO |
| WO 2002076395 | Oct 2002 | WO |
| WO 2003004097 | Jan 2003 | WO |
| WO 2003006427 | Jan 2003 | WO |
| WO 2003015605 | Feb 2003 | WO |
| WO 2003015605 | Feb 2003 | WO |
| WO 2003017989 | Mar 2003 | WO |
| WO03026717 | Apr 2003 | WO |
| WO 2003030659 | Apr 2003 | WO |
| WO 2003041713 | May 2003 | WO |
| WO 2003047636 | Jun 2003 | WO |
| WO 2003047636 | Jun 2003 | WO |
| WO 2003080039 | Oct 2003 | WO |
| WO 2003092763 | Nov 2003 | WO |
| WO 2003095398 | Nov 2003 | WO |
| WO 2003095398 | Nov 2003 | WO |
| WO 2004009066 | Jan 2004 | WO |
| WO 2004009253 | Jan 2004 | WO |
| WO 2004011421 | Feb 2004 | WO |
| WO 2004012874 | Feb 2004 | WO |
| WO 2004037798 | May 2004 | WO |
| WO 2004039313 | May 2004 | WO |
| WO 2004039313 | May 2004 | WO |
| WO 2004060283 | Jul 2004 | WO |
| WO 2004064767 | Aug 2004 | WO |
| WO 2004064767 | Aug 2004 | WO |
| WO 2004087212 | Oct 2004 | WO |
| WO 2004098538 | Nov 2004 | WO |
| WO 2004098538 | Nov 2004 | WO |
| WO 2005003032 | Jan 2005 | WO |
| WO 2005004984 | Jan 2005 | WO |
| WO 2005011575 | Feb 2005 | WO |
| WO 2005011575 | Feb 2005 | WO |
| WO 2005030118 | Apr 2005 | WO |
| WO 2005030118 | Apr 2005 | WO |
| WO 2005030135 | Apr 2005 | WO |
| WO 2005030135 | Apr 2005 | WO |
| WO 2005030147 | Apr 2005 | WO |
| WO 2005030147 | Apr 2005 | WO |
| WO 2005034860 | Apr 2005 | WO |
| WO 2005034860 | Apr 2005 | WO |
| WO 2005039664 | May 2005 | WO |
| WO 2005039664 | May 2005 | WO |
| WO 2005067986 | Jul 2005 | WO |
| WO 2005070006 | Aug 2005 | WO |
| WO 2005070006 | Aug 2005 | WO |
| WO 2005070008 | Aug 2005 | WO |
| WO 2005070008 | Aug 2005 | WO |
| WO 2005070874 | Aug 2005 | WO |
| WO 2005070883 | Aug 2005 | WO |
| WO 2005072819 | Aug 2005 | WO |
| WO 2005077962 | Aug 2005 | WO |
| WO 2005077962 | Aug 2005 | WO |
| WO 2005081752 | Sep 2005 | WO |
| WO 2005081752 | Sep 2005 | WO |
| WO 2005081964 | Sep 2005 | WO |
| WO 2005094913 | Oct 2005 | WO |
| WO 2005102282 | Nov 2005 | WO |
| WO 2005107384 | Nov 2005 | WO |
| WO 2005107384 | Nov 2005 | WO |
| WO 2005112954 | Dec 2005 | WO |
| WO 2005115440 | Dec 2005 | WO |
| WO 2005115440 | Dec 2005 | WO |
| WO 2005120493 | Dec 2005 | WO |
| WO 2006023693 | Mar 2006 | WO |
| WO 2006023693 | Mar 2006 | WO |
| WO 2006037105 | Apr 2006 | WO |
| WO 2006037105 | Apr 2006 | WO |
| WO 2006041855 | Apr 2006 | WO |
| WO 2006041855 | Apr 2006 | WO |
| WO 2006045639 | May 2006 | WO |
| WO 2006055542 | May 2006 | WO |
| WO 2006055542 | May 2006 | WO |
| WO 2006058318 | Jun 2006 | WO |
| WO 2006064056 | Jun 2006 | WO |
| WO 2006066362 | Jun 2006 | WO |
| WO 2006084909 | Aug 2006 | WO |
| WO 2006084910 | Aug 2006 | WO |
| WO 2006084911 | Aug 2006 | WO |
| WO 2006084912 | Aug 2006 | WO |
| WO 2006084913 | Aug 2006 | WO |
| WO 2006084914 | Aug 2006 | WO |
| WO 2006100155 | Aug 2006 | WO |
| WO 2006095193 | Sep 2006 | WO |
| WO 2006095193 | Sep 2006 | WO |
| WO 2006096572 | Sep 2006 | WO |
| WO 2006097348 | Sep 2006 | WO |
| WO 2006099058 | Sep 2006 | WO |
| WO 2006099058 | Sep 2006 | WO |
| WO 2006100154 | Sep 2006 | WO |
| WO 2006100156 | Sep 2006 | WO |
| WO 2006100156 | Sep 2006 | WO |
| WO 2006122960 | Nov 2006 | WO |
| WO 2006122961 | Nov 2006 | WO |
| WO 2006125016 | Nov 2006 | WO |
| WO 2006125262 | Nov 2006 | WO |
| WO 2006127591 | Nov 2006 | WO |
| WO 2006127591 | Nov 2006 | WO |
| WO2006128121 | Nov 2006 | WO |
| WO 2006128742 | Dec 2006 | WO |
| WO 2006128742 | Dec 2006 | WO |
| WO 2006128743 | Dec 2006 | WO |
| WO 2006130982 | Dec 2006 | WO |
| WO 2007003028 | Jan 2007 | WO |
| WO 2007005910 | Jan 2007 | WO |
| WO 2007005910 | Jan 2007 | WO |
| WO 2007007208 | Jan 2007 | WO |
| WO 2007012165 | Feb 2007 | WO |
| WO 2007016677 | Feb 2007 | WO |
| WO 2007016677 | Feb 2007 | WO |
| WO 2007023005 | Mar 2007 | WO |
| WO 2007023396 | Mar 2007 | WO |
| WO 2007024501 | Mar 2007 | WO |
| WO 2007024501 | Mar 2007 | WO |
| WO 2007027859 | Mar 2007 | WO |
| WO 2007028657 | Mar 2007 | WO |
| WO 2007030266 | Mar 2007 | WO |
| WO 2007030266 | Mar 2007 | WO |
| WO 2007050379 | May 2007 | WO |
| WO 2007050379 | May 2007 | WO |
| WO 2007053292 | May 2007 | WO |
| WO 2007053578 | May 2007 | WO |
| WO 2007053578 | May 2007 | WO |
| WO 2007054373 | May 2007 | WO |
| WO 2007054818 | May 2007 | WO |
| WO 2007057763 | May 2007 | WO |
| WO 2007057763 | May 2007 | WO |
| WO 2007059311 | May 2007 | WO |
| WO 2007059311 | May 2007 | WO |
| WO 2007059311 | May 2007 | WO |
| WO 2007064895 | Jun 2007 | WO |
| WO 2007064895 | Jun 2007 | WO |
| WO 2007067477 | Jun 2007 | WO |
| WO 2007084533 | Jul 2007 | WO |
| WO 2007084533 | Jul 2007 | WO |
| WO 2007086884 | Aug 2007 | WO |
| WO 2007086884 | Aug 2007 | WO |
| WO 2007088050 | Aug 2007 | WO |
| WO 2007088050 | Aug 2007 | WO |
| WO 2007088123 | Aug 2007 | WO |
| WO 2007088123 | Aug 2007 | WO |
| WO 2007092284 | Aug 2007 | WO |
| WO 2007092284 | Aug 2007 | WO |
| WO 2007100910 | Sep 2007 | WO |
| WO 2007100910 | Sep 2007 | WO |
| WO 2007103190 | Sep 2007 | WO |
| WO 2007103190 | Sep 2007 | WO |
| WO 2007127725 | Nov 2007 | WO |
| WO 2007127725 | Nov 2007 | WO |
| WO 2007133922 | Nov 2007 | WO |
| WO 2007133922 | Nov 2007 | WO |
| WO 2007143185 | Dec 2007 | WO |
| WO 2007143185 | Dec 2007 | WO |
| WO 2007149437 | Dec 2007 | WO |
| WO 2007149520 | Dec 2007 | WO |
| WO 2007149520 | Dec 2007 | WO |
| WO 2008005313 | Jan 2008 | WO |
| WO 2008005313 | Jan 2008 | WO |
| WO 2008013633 | Jan 2008 | WO |
| WO 2008013633 | Jan 2008 | WO |
| WO 2008020218 | Feb 2008 | WO |
| WO 2008027203 | Mar 2008 | WO |
| WO 2008027203 | Mar 2008 | WO |
| WO 2008032212 | Mar 2008 | WO |
| WO 2008038140 | Apr 2008 | WO |
| WO 2008038147 | Apr 2008 | WO |
| WO 2008062160 | May 2008 | WO |
| WO 2008071242 | Jun 2008 | WO |
| WO 2008088507 | Jul 2008 | WO |
| WO 2008088507 | Jul 2008 | WO |
| WO 2008095841 | Aug 2008 | WO |
| WO 2008095841 | Aug 2008 | WO |
| WO 2008098192 | Aug 2008 | WO |
| WO 2008098192 | Aug 2008 | WO |
| WO 2008100591 | Aug 2008 | WO |
| WO 2008100591 | Aug 2008 | WO |
| WO 2008110872 | Sep 2008 | WO |
| WO 2008112391 | Sep 2008 | WO |
| WO 2008112391 | Sep 2008 | WO |
| WO 2008116497 | Oct 2008 | WO |
| WO 2008116925 | Oct 2008 | WO |
| WO 2008130567 | Oct 2008 | WO |
| WO 2008141416 | Nov 2008 | WO |
| WO 2008150505 | Dec 2008 | WO |
| WO 2008152444 | Dec 2008 | WO |
| WO 2008157393 | Dec 2008 | WO |
| WO 2009007785 | Jan 2009 | WO |
| WO 2009014616 | Jan 2009 | WO |
| WO 2009014829 | Jan 2009 | WO |
| WO 2009014829 | Jan 2009 | WO |
| WO 2009019498 | Feb 2009 | WO |
| WO 2009019498 | Feb 2009 | WO |
| WO 2009019499 | Feb 2009 | WO |
| WO 2009026680 | Mar 2009 | WO |
| WO 2009036571 | Mar 2009 | WO |
| WO 2009049208 | Apr 2009 | WO |
| WO 2009056991 | May 2009 | WO |
| WO 2009064861 | May 2009 | WO |
| WO 2009064861 | May 2009 | WO |
| WO 2009072007 | Jun 2009 | WO |
| WO 2009073643 | Jun 2009 | WO |
| WO 2009073643 | Jun 2009 | WO |
| WO 2009073940 | Jun 2009 | WO |
| WO 2009073940 | Jun 2009 | WO |
| WO 2009080795 | Jul 2009 | WO |
| WO 2009086470 | Jul 2009 | WO |
| WO 2009086470 | Jul 2009 | WO |
| WO 2009087578 | Jul 2009 | WO |
| WO 2009088433 | Jul 2009 | WO |
| WO 2009090495 | Jul 2009 | WO |
| WO 2009098113 | Aug 2009 | WO |
| WO 2009098595 | Aug 2009 | WO |
| WO 2009117182 | Sep 2009 | WO |
| WO 2009117182 | Sep 2009 | WO |
| WO 2009117183 | Sep 2009 | WO |
| WO 2009124379 | Oct 2009 | WO |
| WO 2009131931 | Oct 2009 | WO |
| WO 2009155689 | Dec 2009 | WO |
| WO 2009155690 | Dec 2009 | WO |
| WO 2010002450 | Jan 2010 | WO |
| WO 2010002450 | Jan 2010 | WO |
| WO 2010033242 | Mar 2010 | WO |
| WO 2010033242 | Mar 2010 | WO |
| WO 2010045415 | Apr 2010 | WO |
| WO 2010045465 | Apr 2010 | WO |
| WO 2010048724 | May 2010 | WO |
| WO 2010080213 | Jul 2010 | WO |
| WO 2010080213 | Jul 2010 | WO |
| WO 2010096320 | Aug 2010 | WO |
| WO 2010096320 | Aug 2010 | WO |
| WO 2010114669 | Oct 2010 | WO |
| WO 2010120414 | Oct 2010 | WO |
| WO 2010151505 | Dec 2010 | WO |
| Entry |
|---|
| Notification Concerning Transmittal of International Preliminary Report on Patentability (Chapter 1 of the Patent Cooperation Treaty) corresponding to PCT/US2010/046209; dated Mar. 1, 2012. |
| Shin et al. “Synthesis of Nitric Oxide-releasing Silica Nanoparticles”, Journal of American Chemical Society, 129, pp. 4612-4619, 2007. |
| Barbe et al. “Silica Particles: A Novel Drug-Delivery System”, Advanced Materials, 2004, vol. 16(21), pp. 1959-1965. |
| Dobmeier, K. et al. “Antibacterial Properties of Nitric Oxide-Releasing Sol-Gel Microarrays”, Biomacromolecules, 2004, vol. 5(6), pp. 2493-2495. |
| Farias-Eisner et al. “The Chemistry and Tumoricidal Activity of Nitric Oxide/Hydrogen Peroxide and the Implications to Cell Resistance/Susceptibility”, The Journal of Biological Chemistry, 1996, vol. 271(11), pp. 6144-6151. |
| Hatton, H. et al. “Past, Present, and Future of Periodic Mesoporous Organosilicas the PMOs”, Accounts of Chemical Research, vol. 38, No. 4, Apr. 2005, pp. 305-312. |
| Hetrick, E. et al. “Antibacterial Nitric Oxide-Releasing Xerogels: Cell Viability and Parallel Plate Flow Cell Adhesion Studies”, Biomaterials, 2007, vol. 28(11) pp. 1948-1956. |
| Hetrick, E. et al. “Reducing Implant-Related Infections: Active Release Strategies”, Chem. Soc. Rev., 2006, vol. 35, pp. 780-789. |
| Lin, Hong-Ping et al. “Structural and Morphological Control of Cationic Surfactant-Templated Mesoporous Silica”, Accounts of Chemical Research, vol. 35, No. 11, Nov. 2002, pp. 927-935. |
| Marxer, S. et al. “Sol-gel derived nitric oxide-releasing oxygen sensors”, The Analyst, 2005, vol. 130(2), pp. 206-212. |
| Nablo, B. et al. “Inhibition of Implant-Associated Infections via Nitric Oxide Release”, Biomaterials, 2005, vol. 26(34), pp. 6984-6990. |
| Nablo, B. et al. “Nitric oxide-releasing sol-gels as antibacterial coating for orthopedic implants”, Biomaterials, 2005, vol. 26(8), pp. 917-924. |
| Pulfer, S. et al. “Incorporation of nitric oxide-releasing crosslinked polyethyleneimine microspheres into vascular grafts”, J. Biomed Mater Res, 1997, vol. 37(2),pp. 182-189. |
| Reynolds, M. et al. “Nitric Oxide-Releasing Hydrophobic Polymers: Preparation, Characterization, and Potential Biomedical Applications”, Free Radical Biology & Medicine, 2004, vol. 37(7), pp. 926-936. |
| Shin, J. et al. “Nitric Oxide-Releasing Sol-Gel Particle/Polyurethane Glucose Biosensors”, Anal Chem, 2004, vol. 76, pp. 4543-4549. |
| Stein A. et al. “Hybrid Inorganic Organic Mesoporous Silicates Nanoscopic Reactors Coming of Age”, Advanced Materials, Oct. 2000, 1403-1419. |
| Ashutosh K. et al., “Use of nitric oxide inhalationin chronic obstructive pulmonary disease”, Thorax, 2000;55:109-113. |
| Azizzadeh B. et al., “Nitric Oxide Improve Cisplatin Cytotoxicity in Head and Neck Squamous Cell Carcinoma”, Laryngoscope, 111:Nov. 2001, pp. 1896-1900. |
| Barst R.J. et al., “Clinical perspectives with long-term pulsed inhaled nitric oxide for the treatment of pulmonary arterial hypertension”, Pulmonary Circulation, Apr.-Jun. 2012, vol. 2, No. 2, pp. 139-147. |
| Benz S. et al., “Effect of Nitric Oxide in Ischemia/Reperfusion of the Pancreas”, Journal of Surgical Research, vol. 106, Issue 1, pp. 46-53, Jul. 2002. |
| Bian K. et al., “Vascular System: Role of Nitric Oxide in Cardiovascular Diseases”, The Journal of Clinical Hypertension, vol. 10, No. 4, Apr. 2008, pp. 304-310. |
| Bloch K.D. et al. “Inhaled NO as a therapeutic agent”, Cardiovascular Research, 75, 2007, 339-348. |
| Bonavida B. et al., “Novel therapeutic applications of nitric oxide donors in cancer: Roles in chemo- and immunosensitization to apoptosis and inhibition of metastases”, Nitric Oxide, vol. 19, Issue 2, Sep. 2008, pp. 152-157. |
| Bonavida B. et al., “Therapeutic potential of nitric oxide in cancer”, Drug Resist Updat., Jun. 2006;9(3):157-73, Epub Jul. 5, 2006. |
| Boykin J.V. et al., “HBO mediates increased nitric oxide production associated with wound healing”, Wound Repair and Regeneration, vol. 12, No. 2, Mar.-Apr. 2004. |
| Boykin Jr. J.V., “Wound Nitric Oxide Bioactivity: A Promising Diagnostic Indicator for Diabetic Foot Ulcer Management”, Journal of Wound, Ostomy & Continence Nursing, Jan./Feb. 2010, vol. 37, Issue 1, p. 25-32. |
| Bruch-Gerharz D. et al., “Nitric Oxide in Human Skin: Current Status and Future Prospects”, J. Inves Dermatol, 110:1-7, 1998. |
| Cals-Grierson M.M. et al., “Nitric oxide function in the skin”, Nitric Oxide, vol. 10, Issue 4, Jun. 2004, pp. 179-193. |
| Carlsson S. et al., “Intravesical Nitric Oxide Delivery for Prevention of Catheter-Associated Urinary Tract Infections”, Antimicrob. Agents Chemother. 2005, 49(6):2352. |
| De Groote M.A. et al., “NO Inhibitions: Antimicrobial Properties of Nitric Oxide”, Clinical Infectious Diseases, 1995, 21 (Supplement 2), S162-S165. |
| Fang F., “Mechanisms of Nitric Oxide-related Antimicrobial Activity”, J.Clin. Invest., vol. 99, No. 12, Jun. 1997, 2818-2825. |
| Frederiksen L.J. et al., “Chemosensitization of Cancer in vitro and in vivo by Nitric Oxide Signaling”, Clin Cancer Res. 2007;13:2199-2206. |
| Ghaffari A. et al., “Potential application of gaseous nitric oxide as a topical antimicrobial agent”, Nitric Oxide, vol. 14, Issue 1, Feb. 2006, pp. 21-29. |
| Herman A.G. et al., “Therapeutic potential of nitric oxide donors in the prevention and treatment of atherosclerosis”, European Heart Journal, 2005, 26, 1945-1955. |
| Hirst D. et al., “Targeting nitric oxide for cancer therapy”, Journal of Pharmacy and Pharmacology, 2007, 59: 3-13. |
| Howlin R. et al., “Nitric oxide-mediated dispersal and enhanced antibiotic sensitivity in Pseudomonas aeruginosa biofilms from the cystic fibrosis lung”, Archives of Disease in Childhood, 2011;96:A45. |
| Huerta S. et al., “Nitric oxide donors: Novel cancer therapeutics (Review)”, International Journal of Oncology, 33, 909-927, 2008. |
| Johnson T. A. et al., “Reduced ischemia/reperfusion injury via glutathione-initiated nitric oxide-releasing dendrimers”, Nitric Oxide, 2009, 7 Pages. |
| Jones M.L. et al., “Antimicrobial properties of nitric oxide and its application in antimicrobial formulations and medical devices”, Appl Microbiol Biotechnol, 2010, 88:401-407. |
| Kiziltepe T. et al., “JS-K, a GST-activated nitric oxide generator, induces DNA double-strand breaks, activates DNA damage response pathways, and induces apoptosis in vitro and in vivo in human multiple myeloma cells”, Blood, 2007, 110: 709-718. |
| Lamas S. et al., “Nitric oxide signaling comes of age: 20 years and thriving”, Cardiovascular Research, 75, 2007, 207-209. |
| Liu X. et al., “Nitric Oxide Inhalation Improves Microvascular Flow and Decreases Infarction Size After Myocardial Ischemia and Reperfusion”, Journal of the American College of Cardiology, vol. 50, No. 8, 2007. |
| Luo J. et al., “Nitric oxide: a newly discovered function on wound healing”, Acta Pharmacologica Sinica, Mar. 2005; 26 (3): 259-264. |
| McGrowder D. et al., “Therapeutic Uses of Nitric Oxide-donating Drugs in the Treatment of Cardiovascular Diseases”, International Journal of Pharmacology, 2(4): 366-373, 2006. |
| Napoli C. et al., “Nitric oxide and atherosclerosis: An update”, Nitric Oxide, vol. 15, Issue 4, Dec. 2006, pp. 265-279. |
| Phillips L. et al., “Nitric Oxide Mechanism of Protection in Ischemia and Reperfusion Injury”, Journal of Investigative Surgery, 22, 46-55, 2009. |
| Saavedra J.E. et al., “Esterase-Sensitive Nitric Oxide Donors of the Diazeniumdiolate Family: in Vitro Antileukemic Activity”, J. Med. Chem. 2000, 43, 261-269. |
| Schairer D.O. et al., “The potential of nitric oxide releasing therapies as antimicrobial agents”, Virulence, 3:3, 271-279; May/Jun. 2012. |
| Siriussawakul A. et al. “Role of nitric oxide in hepatic ischemia-reperfusion injury”, World Journal of Gastroenterology, Dec. 28, 2010, 16(48): 6079-6086. |
| Schulz R. et al., “Nitric oxide in myocardial ischemia/reperfusion injury”, Cardiovascular Research, 61, 2004, 402-413. |
| Schwentker A. et al., “Nitric oxide and wound repair: role of cytokines?” Nitric Oxide, vol. 7, Issue 1, Aug. 2002, pp. 1-10. |
| Simeone A.M. et al., “N-(4-Hydroxyphenyl) retinamide and nitric oxide pro-drugs exhibit apoptotic and anti-invasive effects against bone metastatic breast cancer cells” Carcinogenesis, vol. 27, No. 3, pp. 568-577, 2006. |
| Stevens E.V. et al., “Nitric Oxide-Releasing Silica Nanoparticle Inhibition of Ovarian Cancer Cell Growth”, Molecular Pharmaceutics, vol. 7, No. 3, 775-785, 2010. |
| Terpolilli N.A. et al., “Inhalation of Nitric Oxide Prevents Ischemic Brain Damage in Experimental Stroke by Selective Dilatation of Collateral Arterioles”, Circulation Research, 2012;110:727-738. |
| Thomas D.D. et al., “Hypoxic inducible factor 1α, extracellular signal-regulated kinase, and p53 are regulated by distinct threshold concentrations of nitric oxide”, PNAS, Jun. 15, 2004, vol. 101, No. 24, 8894-8899. |
| Weller R. “Nitric oxide donors and the skin: useful therapeutic agents?” Clinical Science, 2003, 105, 533-535. |
| Wink D.A. et al., “The multifaceted roles of nitric oxide in cancer”, Carcinogenesis, vol. 19, No. 5, pp. 711-721, 1998. |
| Witte M.B. et al., “Nitric oxide enhances experimental wound healing in diabetes”, British Journal of Surgery, 2002, 89, 1594-1601. |
| Witte M.B. et al., “Role of nitric oxide in wound repair”, The American Journal of Surgery, vol. 183, Issue 4, pp. 406-412, Apr. 2002. |
| Yetik-Anacak G. et al., “Nitric oxide and the endothelium: History and impact on cardiovascular disease”, Vascular Pharmacology, vol. 45, Issue 5, Nov. 2006, pp. 268-276. |
| Zhu H. et al., “Effects of Nitric Oxide on Skin Burn Wound Healing”, Journal of Burn Care & Research, Sep./Oct. 2008, vol. 29, Issue 5, pp. 804-814. |
| Zhu H. et al., “Nitric Oxide Accelerates the Recovery from Burn Wounds”, World Journal of Surgery, 2007, 31: 624-631. |
| Notification of Transmittal of the International Search Report and the Written Opinion of the International Searching Authority, or the Declaration; International Search Report; Written Opinion of the International Searching Authority corresponding to PCT/US2010/046209; dated May 23, 2011; 13 pages. |
| Norio Iwakiri, et al., Synthesis of amphiphilic polysiloxanes and their properties for formation of nano-aggregates, Colloid Polym Sci, 2009, 287:577-582. |
| Amadeu et al., “Nitric Oxide Donor Improves Healing if Applied on Inflammatory and Proliferative Disease,” J. Surgical Research 149: 84-93 (2008). |
| Barraud, N., et al., “Involvement of Nitric Oxide in Biofilm Dispersal of Pseudomonas aeruginosa,” Journal of Bacteriology, 2006, vol. 188(21), pp. 7344-7353. |
| Bohl Masters et al., “Effects of nitric oxide releasing vinyl poly(vinyl alcohol) hydrogel dressings on dermal wound healing in diabetic mice”, Wound Repair and Regeneration 10(5): 286-294 (2002). |
| Brennan et al., “The Role of Nitric Oxide in Oral Diseases”, Archives of Oral Biology, 2003, vol. 48, pp. 93-100. |
| Coban, A., et al., “The Effect of Nitric Oxide Combined with Fluoroquinolones against Salmonellaenterica Serovar Typhimurium in Vitro,” Mem Inst Oswaldo Cruz, Rio de Janeiro, 2003, vol. 98(3), pp. 419-423. |
| Gupta, R., et al., “Bioactive materials for biomedical applications using sol-gel technology,” Biomed Mater., 2008, vol. 3, pp. 1-15. |
| Hetrick et al., “Anti-biofilm efficacy of nitric oxide-releasing silica nanoparticles”, Biomaterials 30:2782-2789 (2009). |
| Hetrick et al., “Bactericidal Efficacy of Nitric Oxide-Releasing Silica Nanoparticles,” Acsnano, 2008, vol. 2(2), pp. 235-246. |
| Hrabie et al., “Chemistry of the nitric oxide-releasing diazeniumdiolate (“nitrosohydroxylamine”) functional group and its oxygen-substituted derivatives,” Chem Rev. 2002, 102, p. 1135-1154. |
| International Search Report and Written Opinion for corresponding PCT application No. PCT/US2009/005643, dated May 24, 2010. |
| Living Water Acid-Alkaline Balance http://www.livingwaterhealthsolutions.com/Articles/alkalize.php Accessed online Nov. 3, 2011. |
| McElhaney-Feser, G., et al., “Synergy of Nitric Oxide and Azoles against Candida Species in Vitro,” Antimicrobial Agents and Chemotherapy, 1998, vol. 42(9), pp. 2342-2346. |
| Notification Concerning Transmittal of International Preliminary Report on Patentability for corresponding PCT Application No. PCT/US2009/005643, dated Apr. 28, 2011. |
| Notification of the Transmittal of the International Search Report and Written Opinion of the International Searching Authority, or the Declaration corresponding to International Application No. PCT/US2010/046173 dated Dec. 6, 2010. |
| Notification of the Transmittal of the International Search Report and the Written Opinion of the International Searchning Authority, or the Declaration; International Search Report; Written Opinion of the International Searching Authority corresponding to PCT/US2010/052460; dated Jan. 24, 2011; 10 pages. |
| Robson, MC, “Wound Infection. A Failure of Wound Healing Caused by an Imbalance of Bacteria,” Surg Clin North Amer, Jun. 1997; 77(3): 637-50. |
| Saaral, NY, “The Equilibrium Between Endothelin-1/Nitric Oxide in Acne Vulgaris,” Istanbul Tip Fakultesi Dergisi Cilt, 2008, 71(4). |
| Salivary pH Testing https://allicincenter.com/pdf/ph_testing.pdf Accessed online Nov. 3, 2011. |
| Sato, EF et al., J. Clin. Biochem. Nutr. (Pub Online Dec. 28, 2007), 42; pp. 8-13. |
| Schaffer, MR, et al., “Diabetes-impaired healing and reduced wound nitric oxide synthesis: a possible pathophysiologic correlation”. Surgery, May 1997; 121(5):513-9. |
| Shi, HP, et al., “The role of iNOS in wound healing”. Surgery, vol. 130, Issue 2, Aug. 2001; pp. 225-229. |
| Slowing, I.I., et al. Adv. Drug Del. Rev. (Aug. 2008), 60; pp. 1278-1288. |
| Stasko, N., et al., “Dendrimers as a Scaffold for Nitric Oxide Release,” J. Am. Chem. Soc., 2006, vol. 128, pp. 8265-8271. |
| Summersgill, J., et al., “Killing of Legionella pneumophila by nitric oxide in γ-interferon-activated macrophages,” Journal of Leukocyte Biology, 1992, vol. 52, p. 625-629. |
| Tang, X., et al., “Synthesis of Beta-Lactamase Activated Nitric Oxide Donors,” Biorgania & Medicinal Chemistry Letters, 2003, vol. 13, pp. 1687-1690. |
| Zhu, D., et al., “Corrosion protection of metals by water-based silane mixtures of bis-[trimethosysilylpropyl]amine and vinyltriacetoxysilane,” Progress in Organic Coatings, 2004, vol. 49, pp. 42-53. |
| Rothrock et al. “Synthesis of Nitric Oxide-Releasing Gold Nanoparticles”, J. Am. Chem. Soc. 127:9362-9363 (2005). |
| Zhang et al. “Nitric Oxide-Releasing Fumed Silica Particles: Synthesis, Characterization, and Biomedical Application”, J. Am. Chem. Soc. 125:5015-5024 (2003). |
| Office Action corresponding to Canadian Application No, 2,606,565 dated Mar. 27, 2013. |
| Supplementary European Search Report corresponding to European Application No. 06771501.1 dated Dec. 14, 2012. |
| Notification of the Transmittal of the International Search Report and the Written Opinion of the International Searching Authority, or the Declaration; International Search Report; Written Opinion of the International Searching Authority corresponding to PCT/US2010/052460; dated Jan. 24, 2011; 10 pages. |
| Saaral, NY, “The Equilibrium Between Endothelin-1/Nitric Oxide in Acne Vulgaris,” Istanbul Tip Falcultesi Dergisi Cilt, 2008, 71(4). |
| Slowing, I.I., et al. Adv. Drug Del. Rev.(Aug. 2008), 60; pp. 1278-1288. |
| Stasko, N., et al., “Dendrimers as a Scaffold for Nitric Oxide Release,” J. Am. Chem Soc., 2006, vol. 128, pp. 8265-8271. |
| Chinese Office Action Corresponding to Chinese Patent Application No. 201080047383.8; dated Oct. 17, 2013, 2013; 23 Pages. |
| Chinese Office Action Corresponding to Chinese Patent Application No. 201080047383.8; dated Jun. 10, 2014. |
| Davies et al., “Chemistry of the Diazeniumdiolates. 2. Kinetics and Mechanism of Dissociation to Nitric Oxide in Aqueous Solution”, Journal of the American Chemical Society, 2001, 123: 5473-5481. |
| European Examination Report Corresponding to European Patent Application No. 10 747 385.2; dated Sep. 2, 2014. |
| Office Action corresponding to Chinese Patent Application No. 201080047383.8; dated Dec. 2, 2014. |
| Office Action corresponding to Chinese Patent Application No. 201080047383.8; dated May 13, 2015. |
| Sangster, James “Octanol-Water Partition Coefficients of Simple Organic Compounds” Journal of Physical and Chemical Reference Data 18(3):1111-1227 (1989). |
| Office Action corresponding to Chinese Patent Application No. 201080047383.8; dated Oct. 27, 2015 (19 pages). |
| Examination Report corresponding to European Patent Application No. 10747385.2 (5 pages) (dated Jul. 21, 2016). |
| Boykin et al. “Hbo Mediates Increased Nitric Oxide Production Associated With Wound Healing” Wound Repaid and Regeneration 12(2):A15 (Abstract 054) (2004). |
| Number | Date | Country | |
|---|---|---|---|
| 20120136323 A1 | May 2012 | US |
| Number | Date | Country | |
|---|---|---|---|
| 61235927 | Aug 2009 | US | |
| 61235948 | Aug 2009 | US |
