The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 31, 2013 is named SequenceListing.txt and is 699 bytes in size.
(a) Field of the Invention
The present invention relates to the composition of molecular assemblies in liquid solution that enables temperature-controlled pH-dependant formation of biopolymeric gels, such as polysaccharide-based, and methods of preparation thereof.
(b) Description of Prior Art
Biopolymers and macromolecules are attractive materials for the preparation and design of self-gelling and/or auto-assembling systems. Numerous attempts tend to develop such systems on the basis of polysaccharides and polypeptides.
In situ formed gels were also proposed with ionic polysaccharides. A composition can be used as a medical device for drug delivery, the application of a diagnostic agent, or the prevention of post-operative adhesions, and is composed of an aqueous liquid vehicle which is capable of being gelled in situ. It includes at least one ionic polysaccharide, at least one film forming polymer, and a medicament or pharmaceutical agent, water, and optionally, a counter-ion capable of gelating the ionic polysaccharide. However, the gelation is reached by interaction between the ionic polysaccharide and the film-forming polymer, or by counter-ion induced cross-linking of the ionic polysaccharide. Other in situ forming gels are based upon polyoxyalkylene composition or polyoxyalkylene/polysaccharide mixture or alginate/cation mixture in situ.
It would be highly desirable to be provided with a biopolymeric gel that is formed while excluding any organic solvent, any organic monomers, any ionic or covalent cross-linking that may be potentially toxic or induce a reduced biological compatibility.
It would be highly desirable to be provided with a biopolymeric gel that is formed by stimulus-induced free interactions between biologically acceptable and well-recognized molecules.
It would be highly desirable to be provided with, a temperature-controlled pH-dependant formed biopolymeric gels that could be used to encapsulate cells and cellular material while retaining their biological activity.
It would be highly desirable to be provided with such gels, which would retain its solid or gel state at the physiological temperature or 37° C.
One aim of the present invention is to provide a way allowing the preparation of a neutral clear liquid solution of a pH-controlled acid-soluble biopolymer while avoiding any unwanted precipitation or heterogeneous gelation.
A second major aim of the present invention is to provide a neutral clear liquid solution of a pH-controlled acid-soluble biopolymer that will thermally form solid homogeneous gels at a temperature close to the physiological temperature.
Another aim is to provide temperature-controlled pH-dependant formed gels, which could be used to encapsulate cells and cellular material while retaining their biological activity.
A further aim of the present invention is to provide gels that would retain its solid or gel state at the physiological temperature or 37° C.
Still one aim of the present invention is to provide a method for the preparation of such gels.
In accordance with the present invention, there is provided a biopolymeric liquid aqueous composition for producing self-gelling systems and gels, which comprises:
The composition can be prepared from organic and/or inorganic acids, such as hydrochloric acid, citric acid, ascorbic acid, lactic acid, lactobionic acid, acetic acid, salicylic acid, formic acid, glutamic acid, phosphoric acid, orthophosphoric acid, or glycerophosphoric acid, or a mixture thereof.
The biopolymer preferably comprises a pH-gelling acid-soluble polysaccharide, polypeptidic or poly(amino acids), or synthetic polymer, such as a solution of chitosan, modified chitosan or chitosan derivative, the solution of chitosan being cationic and bearing amino groups.
The molecule, residue or sequence may be an organic salt selected from the group consisting of mono-phosphate salt, mono-sulfonate salt, mono-sulfate salt and mono-carboxylate salt.
Alternatively, the molecule, residue or sequence may be a salt of polyol selected from the group consisting of mono-phosphate dibasic salt, mono-sulfonate salt, mono-sulfate salt and mono-carboxylate salt of polyol, said polyol being selected from the group consisting of glycerol, histidinol, acetol, diethylstil-bestrol, indole-glycerol, sorbitol, ribitol, xylitol, arabinitol, erythritol, inositol, mannitol, glucitol, palmitoyl-glycerol, linoleoyl-glycerol, oleoyl-glycerol, and arachidonoyl-glycerol, or a mixture thereof.
The glycerol may also be selected from the group consisting of glycerol-2-phosphate, sn-glycerol 3-phosphate and L-glycerol-3-phosphate salt, or a mixture thereof.
In a further embodiment, the molecule, residue or sequence is a salt of a sugar selected from the group consisting of mono-phosphate dibasic salt, mono-sulfonate salt, mono-sulfate salt and mono-carboxylate salt of a sugar, said sugar being selected from the group consisting of fructose, galactose, ribose, glucose, xylose, rhamnulose, sorbose, erythrulose, deoxy-ribose, ketose, mannose, arabinose, fuculose, fructopyranose, ketoglucose, sedoheptulose, trehalose, tagatose, sucrose, allose, threose, xylulose, hexose, methylthio-ribose, and methylthio-deoxy-ribulose, or a mixture thereof.
The molecule, residue or sequence may be selected from the group consisting of sodium, magnesium or iron salt of glycerol-2-phosphate, sn-glycerol-3-phosphate and L-glycerol-3-phosphate, glucose-1-phosphate, glucose-6-phosphate, fructose-1-phosphate and fructose-6-phosphate, or a mixture thereof.
The molecule, residue or sequence is preferably a sodium, magnesium or iron salt selected from the group consisting of N-[carbamoylmethyl]-2-aminoethane sulfonate (ACES), N,N-bis[2-hydroxyethyl]-2-aminoethane sulfonate (BES), 3-[N,N-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulfonate (DIPSO), N-[2-hydroxyethyl]piperazine-N′-3-propane-sulfonate (EPPS), N-[2-hydroxyethyl]piperazine-N′-4-butane-sulfonate (HEPBS), N-[2-hydroxyethyl]piperazine-N′-3-propanesulfonate (HEPES), N-[2-hydroxyethyl]piperazine-N′-2-hydroxypropanesulfonate (HEPSO), 2-[N-morpholino]ethanesulfonate (MES), 4-[N-morpholino]butanesulfonate (MOBS), 3-[N-morpholino]butanesulfonate (MOPS), 3-[N-morpholino]-2-hydroxypropanesulfonate (MOPSO), piperazine-N,N′-bis[2-ethanesulfonate] (PIPES), piperazine-N,N′-bis[2-hydroxypropanesulfonate] (POPSO), 3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonate (TAPSO), and N-tris[hydroxymethyl]methyl-2-minoethanesulfonate (TES), and derivatives or mixtures thereof.
The molecule, residue or sequence is preferably selected from the group consisting of N,N-bis[hydroxyethyl]glycine (BICINE), bis[2-hydroxyethyl]iminotris[hydroxymethyl]methane (BIS-TRIS), Glycyl-glycine (GLY-GLY), Triethanolamine (TEA), N-tris[hydroxymethyl]methylglycine (TRICINE), and Tris[hydroxymethyl]aminomethane (TRIZMA), and derivatives or mixtures thereof.
Still in another embodiment, the molecule, residue or sequence has either one acid group and at least one amino group, or more amino groups than acid groups. The molecule, residue or sequence may also be an amino-acid residue, an amino-acid sequence or a poly(amino acids) having a basic character and a pKa between 6.0 and 8.4.
Examples of amino acid residue can be histidine (HIS), arginine (ARG), lysine (LYS), asparagine (ASP), and glutamine (GLN), or a mixture thereof. The amino acid residue may further be modified with a radical acetyl, t-butyl, benzyl, benzoyl, ethyl, formyl, or methyl.
The molecule, residue or sequence is alternatively a sequence, derivative or polymer of at least one amino acid selected from the group consisting of alanine (ALA), histidine (HIS), arginine (ARG), lysine (LYS), aspartic acid (ASP), glutamine (GLN), glycine (GLY), hydroxyproline (HYP), isoleucine (ILE), leucine (LEU), norleucine (NLE), phenylalanine (PHE), proline (PRO), serine (SER), threonine (THR), tyrosine (TYR), and valine (VAL).
Preferably, the composition further comprises at least one other water-soluble polymer, such as collagen, methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl propylcellulose, hydroxymethyl propyl cellulose, polyethylene oxide, polypropylene oxide, poly(ethylene oxide-co-propylene oxide) copolymers, poly(ethylene oxide-co-propylene oxide-co-ethylene oxide) copolymers, polyvinyl alcohol, or polycaprolactone diols, and derivatives or mixtures thereof.
The composition of the present invention may further comprises a solid particulate or a water-soluble additive, such as a drug or a pharmaceutical agent, microorganisms, plant cells, animal cells or human cells dispersed therein.
The composition of the present invention may be used as a carrier for delivering a pharmaceutical agent in situ.
Still in accordance with the present invention, there is provided a method for preparing a composition as defined above. The method comprises the steps of:
a) dissolving a pH-gelling acid-soluble biopolymer within an aqueous acidic solution of a pH from about 1.0 to about 5.0 to obtain an aqueous biopolymer composition having a concentration of 0.1 to 5% by weight of the biopolymer;
b) dissolving 0.1 to 10% by weight of a water-soluble molecule having a basic character and a pKa between 6.0 and 8.4, or a water-soluble residue or sequence of the molecule having a basic character and pKa between 6.0 and 8.4, within the aqueous biopolymer composition to obtain a clear liquid formulation with a pH ranging between 5.8 and 7.4;
c) heating the liquid formulation at a temperature above 30° C. to obtain a solid gel, the gel having a pH from about 5.8 to about 7.4.
The composition of the present invention may be used in cosmetics, pharmacology, medicine and/or surgery, into an implantable device or an implant for repair, reconstruction and/or replacement of tissues and/or organs, as an implantable, transdermal or dermatological drug delivery system, as an opthalmological implant or a drug delivery system, or in cells-loaded artificial matrices for engineering and culture of bioengineered hybrid materials and tissue equivalents.
The composition may be loaded with cells selected from the group consisting of chondrocytes (articular cartilage), fibrochondrocytes (meniscus), ligament fibroblasts (ligament), skin fibroblasts (skin), tenocytes (tendons), myofibroblasts (muscle), mesenchymal stem cells and keratinocytes (skin). Such composition may be used in culturing and engineering of artificial articular cartilage and cartilaginous tissues and organs, either for surgical or laboratory testing applications.
The composition of the present invention may also be used in processing and engineering of living artificial substitutes for ligaments, tendons, skin, bone muscles and any metabolic organs, either for surgical or laboratory testing applications, in living substitutes for the replacement of articular cartilages, fibrocartilages, cartilaginous organs, ligaments, tendons, bone tissues or skin, to induce an ectopic formation of fibrocartilage-like or cartilage-like tissues, as an injectable or implantable gel biomaterial which acts as supports, carriers, reconstructive devices or substitutes for the formation in situ of bone-like, fibrocartilage-like or cartilage-like tissues, and/or in cosmetics, pharmacology, medicine and/or surgery.
For the purpose of the present invention the following terms and expressions are defined below.
The term “gelating temperature” is intended to mean any temperature ranging from about 20° C. to about 80° C., preferably between 30° C. and 60° C., and more preferably at about the physiological temperature or 37° C.
The term “pH-controlled acid-soluble biopolymer” refers to a biological polymer that is solubilized in an acidic aqueous medium, and precipitates or gels heterogeneously when the pH is increased. For example, chitosan is dissolved in acid/water solution at pH about 4.0, and precipitates or gels heterogeneously when the chitosan solution is neutralized at pHs above 6.2.
The expression “three-dimensional” refers herein to the fact that the polymeric solution is simultaneously gelated and shaped by the mold wherein the solution was initially poured. Gels can be produced in glass or plastic bechers, dishes, tubes or between two plates so as to obtain any expected shapes.
The expression “in situ gelation” refers herein to the formation of gels by injecting the liquid solution within specific sites of mammalian or human environments, e.g. any tissues (muscles, bone, ligaments, cartilages) and organs. Gelation in situ allows complete and precise filling of tissue defects or body cavities. The gelation of biopolymer mixture is induced by the physiological temperature.
The expression “endothermal gelation” refers herein to the thermal mechanism of the solution, which enables the solution to gelate upon standing at the desired temperature. Induction of sol to gel transitions of systems requires energy via, for example, the temperature.
The expression “residue” refers herein to a series of biochemical molecules having a common specific chemical function. Example: the amino acid residues.
The expression “sequence” refers herein to the association of two or several molecules or residues. Example: a sequence of amino acid residues (LYS-ASP-PRO-GLY-LYS).
The expression “basic character” refers herein to the ability of a chemical molecule in aqueous solution to capture protons (H+), thus leading to an increase in pH.
The expression “cells or cellular matters” refers herein to living biologicals, such as isolated cells, cellular dispersion, cell aggregates, cell spheroids or cells adhered to solid microspheres particles, that are encapsulated within the gels.
It is assumed herein that any pH-gelling acid-soluble biopolymers or polymers would behave similarly. As a consequence, the term “biopolymer” in the present invention may be replaced by the term “polymer”, “polypeptide”, “poly(amino acids)”.
The present invention include method of forming different gelated materials, those materials being either molded (customized shapes, tubes, membranes, films . . . ) or formed in situ within biological environments (filling of tissue defects).
In a preferred embodiment, the self-gelling biopolymer aqueous solution has a pH above that for normal precipitation, and turn into solid gel upon thermal stimulation. This biopolymer gel can be used as a carrier for drugs or as a non-living therapeutics delivery systems, as substituting materials for tissues and organs and as encapsulants for living cells or microorganisms. Gel matrices are rapidly formed at temperatures between 30 to 60° C. Such aqueous systems are used as injectable filling materials, injected and gelated in situ for filling and repairing tissue defects.
In a second preferred embodiment, pH-dependant gelling acid water-soluble biopolymers and derivatives having a sufficient hydrophilicity are selected for preparing temperature-stimulated gels.
Biopolymeric gels can be applied to surgical reconstructive and regeneration uses and drug delivery purposes. They provide thermally reversible or irreversible bioerodible polymeric gels with biologically well-known and compatible components for a broad range of medical/biotechnological applications.
In accordance with the present invention there is proposed a new gelation mechanism that combines hydrogen bonding, electrostatic interactions and hydrophilic/hydrophobic interactions. It can only be achieved through complex interactions between biological macromolecules or synthetic polymers, water molecules and specific biochemical molecules having special actions.
In accordance with the present invention, the concerned biopolymer should be insoluble in water under neutral conditions pH=7.
A method is disclosed for preparing a composition which comprises the steps a) of dissolving a pH-gelling acid-soluble biopolymer within an aqueous acidic solution of a pH from about 1.0 to about 5.0 to obtain an aqueous biopolymer composition having a concentration of 0.1 to 5% by weight of said biopolymer, and b) dissolving 0.1 to 10% by weight of a water-soluble molecule having a moderate basic character, or any water-soluble sequence of said molecule, within said aqueous biopolymer composition to obtain a clear liquid formulation with a pH ranging between 6.5 and 7.4. The final step is the heating of liquid formulation at a temperature above 30° C. to obtain a solid gel, wherein said gel has a concentration of 0.1 to 5.0% by weight of said biopolymer, and a concentration of 0.1 to 10% by weight of said molecule, and has a pH from about 6.4 to about 7.4.
The aqueous acidic solution is prepared from organic or inorganic acids that are selected from the group consisting of acetic acid, ascorbic acid, glutamic acid, lactic acid, lactobionic acid, salicylic acid, phosphoric acid, hydrochloric acid, propionic acid, formic acid, and a mixture thereof. Solubilization of pH-controlled acid-soluble biopolymers in aqueous solution requires acidic aqueous solutions having a pH ranging from 1.0 to 5.0.
The selected biopolymer is a pH-gelling acid-soluble polysaccharide, polypeptidic or poly(amino acids), or synthetic polymer.
The preferred polysaccharide is selected from a group comprising chitosan, modified chitosan or chitosan derivative, said chitosan biopolymer being cationic and bearing amino groups.
The acid-soluble polypeptide is selected from collageneous proteins, preferentially collagen.
A second polymer, selected from a groups comprising collagen, methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl propylcellulose, hydroxymethyl propyl cellulose, polyethylene oxide, polypropylene oxide, poly(ethylene oxide-co-propylene oxide) copolymers, poly(ethylene oxide-co-propylene oxide-co-ethylene oxide) copolymers, polyvinyl alcohol, polycaprolactone diols, and derivatives, and any mixture thereof, can be incorporated within the biopolymeric solution.
The admixed molecule is required to play a double role: 1) to increase the pH within the biopolymeric solution up the physiological conditions, and 2) to prevent the immediate gelation or aggregation. The required molecule, preferentially selected from organic salts and amino-acids, should have a moderate basic character and a pKa between 6.0 and 7.6. Typically, the selected molecule should have a great sensitivity in terms of hydrophilicity/hydrophobicity (hydrophobic hydration and dehydration) and thermal sensitivity. Such effects are based upon a competition for hydration between apolar and polar groups of said molecule, which enables the design of molecular machines by free energy conversion.
Other preferred molecules, residues or sequences are organic salts selected from mono-phosphate salts, mono-sulfonate salts, mono-sulfate salts or mono-carboxylate salts; said organic salts being water-soluble and having a basic character and a pKa between 6.0 and 7.6.
The organic salt is preferably a salt of polyol or sugar selected from mono-phosphate dibasic salts, mono-sulfonate salts, mono-sulfate salts or mono-carboxylate salts of polyol, said polyol being selected from the group consisting of glycerol, comprising glycerol-2-phosphate, sn-glycerol 3-phosphate and L-glycerol-3-phosphate salts, and any mixture thereof. Salt of polyol or sugar are known to greatly modify the behavior of biopolymeric acidic aqueous solutions.
The salt of polyol is preferably selected from mono-phosphate dibasic salts, mono-sulfonate salts, mono-sulfate salts or mono-carboxylate salts of polyol, said polyol being selected from a group of polyols comprising histidinol, acetol, diethylstil-bestrol, indole-glycerol, sorbitol, ribitol, xylitol, arabinitol, erythritol, inositol, mannitol, glucitol, palmitoyl-glycerol, linoleoyl-glycerol, oleoyl-glycerol, arachidonoyl-glycerol, and any mixture thereof.
The salt of sugar is preferably selected from mono-phosphate dibasic salts, mono-sulfonate salts, mono-sulfate salts or mono-carboxylate salts of sugar, said sugar being selected from a group of sugars consisting of fructose, galactose, ribose, glucose, xylose, rhamnulose, sorbose, erythrulose, deoxy-ribose, ketose, mannose, arabinose, fuculose, fructopyranose, ketoglucose, sedoheptulose, trehalose, tagatose, sucrose, allose, threose, xylulose, hexose, methylthio-ribose, methylthio-deoxy-ribulose, and any mixture thereof.
Polyols are frequently added to compositions for improving gel properties. Sorbitol and mannitol are currently used as tonicity enhancing agents. Glycerol and polyethylene glycol are proposed as plasticizers. Polyols (-ol: glycerol, sorbitol . . . ) and sugars (-ose: fructose, glucose, galactose . . . ) were used as thermal stabilizing agents for proteins in solutions Depending on the selected molecules, they were found to make or break structuring of water, create hydrogen bonding, electrostatic or hydrophobic interacting, and present endothermic transitions Polyols and sugars stabilize proteins to heat denaturation through their structuring effect on water and the strengthen of hydrophobic interactions.
The molecule, residue or sequence is preferably a salt selected in a group comprising N-[carbamoylmethyl]-2-aminoethane sulfonate (ACES), N,N-bis[2-hydroxyethyl]-2-aminoethane sulfonate (BES), 3-[N,N-bis(2-hydroxy-ethyl)amino]-2-hydroxypropanesulfonate (DIPSO), N-[2-hydroxyethyl]piperazine-N′-3-propanesulfonate (HEPES), 2-[N-morpholino]ethanesulfonate (MES), 4-[N-morpholino]butanesulfonate (MOBS), 3-[N-morpholino]butanesulfonate (MOPS), 3-[N-morpholino]-2-hydroxypropanesulfonate (MOPSO), 3-[N-tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonate (TAPSO), N-tris[hydroxymethyl]methyl-2-minoethanesulfonate (TES), bis[2-hydroxy-ethyl]iminotris[hydroxymethyl]methane (BIS-TRIS), 3-morpholino 1-1-propane diol, and derivatives, and any mixture thereof.
The molecule, residue or sequence is preferably selected from amino-acid residues, amino-acid sequences or poly(amino acids) having a basic character and a pKa between 6.0 and 7.6, preferentially histidine (His).
The molecule, residue or sequence is preferably a sequence, derivative or oligomer of amino acids including alanine, (ALA), histidine (HIS), arginine (ARG), lysine (LYS), aspartic acid (ASP), glutamine (GLN), glycine (GLY), hydroxyproline (HYP), isoleucine (ILE), leucine (LEU), norleucine (NLE), phenylalanine (PHE), proline (PRO), serine (SER), threonine (THR), tyrosine (TYR), and valine (VAL).
A pharmaceutical or bioactive agent can be added to the liquid biopolymer containing solution of step a) or b). It can be highly soluble, sparingly soluble or non-soluble in the aqueous formulation. Solid particulate additives such as non-polymeric microspheres or nanospheres, mineral or ceramic granules or powders, can be added to the biopolymer solution of step a) or b).
The mixture can be dispensed for gelation into a desired receiver, either in a mold or within a tissue, organ or body cavity. It can be kept in a stable ungelled liquid form at a temperature ranging from about 0° C. to about 20° C. The solidifying temperature is ranging from about 37° C. to about 60° C., and preferably about 37° C.
Practically, the mixture is introduced within an animal or human body by injection or endoscopic administration, and gelled in situ at a temperature of about 37° C.
Table 1 below provides composition of some preferred examples for buffering/gelling agents with a 2% w/v chitosan solution (deacetylation 85%).
Formation of Biopolymeric Gels
A selected biopolymer in powder form is dissolved in an aqueous acidic solution until the occurrence of a clear solution is obtained. The proportion of biopolymer varies from 0.5 to 10.0% w/v, preferentially from 1.0 to 3.0% w/v. The pH of the aqueous biopolymer solution ranges from 4.0 to 5.5. Aqueous biopolymer solutions can be sterilized either by autoclaving or filtration with in-line sterile filters (0.22 micrometer). Freshly-prepared aqueous biopolymer solutions are stored preferably at low positive temperature (4° C.). The added molecule with a moderate basic character is dissolved in water, then admixed to the aqueous biopolymer solution at a temperatures ranging from 4 to 15° C., preferably 10° C. When a clear homogeneous aqueous solution with a pH ranging from 5.8 to 7.0 is attained, the said solution is poured into the desired receiver, and hold to appropriate temperature to gel.
The nature of the acid that is used for the acidic biopolymer solutions does not influence fundamentally the sol to gel transition of the system. The final pH within the solution is dependent upon the pH of the water/acid solution as well as the biopolymer and molecule concentrations. As the biopolymer and molecule are two basic components, they tend to increase the pH of the acidic solution wherein they are dissolved. Concentrations in this biopolymer and molecule can be balanced to reach the appropriate pH of the solution, while taking into consideration the solubility limit of both components, and particularly the one of biopolymer.
In Situ Formation of Gels
The selected molecule tested to be incorporated in the polymeric solution was histidine, but similar results were obtained with other amino acids or synthetic molecules having similar functions and basic character. In situ gelation of the biopolymer solution can be conducted by dispensing the solution from a hypodermic syringe. If needed, the solution may be pre-gelated (initiate the thermal gelation) by keeping the syringe and biopolymer solution at desired temperature, ideally 37° C., until the first signs of gelation appear. The ready-to-gel biopolymer mixture is then administrated so as to fill tissue defects or cavities and complete in situ the gelation process (at 37° C.). A needle having a gauge of 20 and below is ideal material for injection of such gel solution. Body cavities and tissue defects act as recipients for the solution, but the liquid materials remain in an open aqueous environment. The conformability and diffusability of the biopolymer solutions is dependent upon the solution and material properties. Increased viscosity results in formation in situ of more compact and less conformable gels.
Therapeutic Use and Other Uses of Biopolymeric Gels
Such a biopolymeric gel as previously described is an ideal material for drug delivery system. Such a in situ gel-like forming vehicle, wherein a solid particulate or water-soluble additive is incorporated prior to the gelation, can be administrated topically, directly to the body site to be treated or diagnosed. Anti-bacterial, anti-fungal, steroidal or non-steroidal anti-inflammatory, anti-cancer, anti-fibrosis, anti-viral, anti-glucoma, miotic and anti-cholinergies, anti-psychotic, anti-histaminic and decongestant, anesthetic and anti-parasitic agents may be incorporated within the composition and gel. In a similar fashion, non-living pharmaceutical agents may be incorporated within the composition or gel for restorative, re-constructive or regenerative purposes.
Living microorganisms, plant cells, animal cells or human cells may be entrapped identically within the biopolymer gel by introduction prior to the gelation. The cells or micro-organisms loaded gels may be applied to biotechnological purposes in medicine or in other industrial areas. Biopolymer in situ forming gels can be formed sub-cutaneously, intramuscularly, intra-peritoneally or within biological connective tissues, bone defects, fractures, articular cavities, body conduits or cavities, eye cul-de-sac, solid tumor vasculatures, etc. . . .
The present invention will be more readily understood by referring to the following examples, which are given to illustrate the invention rather than to limit its scope.
This example shows typical experiments of acidic biopolymer solutions neutralized with an organo-phosphate, preferentially glycerophosphate (GP), and transformed to gel upon standing at 37° C.
In typical experiment, 200 mg of chitosan 85% deacetylated was dissolved in 8.5 mL of aqueous HCl solution (0.1M). The chitosan solution, so obtained had a pH of about 5.0, was cooled down to around 4° C. Then 800 mg of β-glycerophosphate disodium salt pentahydrate dissolved in 1.5 mL of water were added slowly to the chitosan solution, while maintaining the cold temperature. The pH of the resulting homogeneous and clear liquid mixture become 7.1. This mixture was disposed in a glass scintillation vial in the incubator at 37° C. for 2 hours, enough time to achieve bulk gelation process.
Similar results were obtained when the β-glycerophosphate disodium salt was replaced by the α-glycerophosphate disodium salt.
Collagen was isolated from knee joint cartilage of calf, and was made mainly of type II collagen. An aqueous solution of collagen (2% w/v) was prepared by dissolving 0.2 g of collagen in 8.5 ml of an acetic acid solution with a pH about 3.6. Once a clear solution was obtained, it was cooled down to about 4° C., and then a cold solution of 800 mg of β-glycerophosphate disodium salt pentahydrate in 1.5 mL of water was added under continuous stirring. When the resulting neutral solution (pH=7.2) appeared quite homogeneous and clear, it was poured in a Petri dish and placed at 37° C. A homogeneous uniform gel formed within 1 hour.
Collagen (100 mg) of the same origin (Example 1, Experiment 2) was first dissolved in 10 ml of an acetic acid solution (0.1M). Then 100 mg of Chitosan was added to the resulting solution and stirred until all chitosan was completely dissolved. After the whole system was cooled down to around 4° C., and 800 mg of β-glycerophosphate disodium salt, dissolved in 1.5 ml of water, was added under continuous stirring. Once the resulting neutral solution (pH=7.2) was perfectly homogeneous and clear, it was poured in a Petri dish and placed at 37° C. The gel formed within 1 hour.
This example shows the typical experiments of acidic biopolymer solutions neutralized with organo-sulfonate, preferentially N,N-bis[2-hydroxyethyl]-2-aminoethane sulfonate (BES), and transformed to gel upon standing at 37° C.
In this experiment, 200 mg of chitosan 85% deacetylated was dissolved in 8.5 mL of aqueous HCl solution (0.1M). The chitosan solution, so obtained had a pH of about 5.0, was cooled down to around 4° C. Then 200 mg of BES in form of sodium salt was dissolved in 1.5 mL of cold water and added slowly to the cold chitosan solution under vigorous stirring. The pH of the resulting homogeneous and clear solution increases to about 7.17. This solution was disposed in a glass scintillation vial in the incubator at 37° C. Bulk gelation occurs in 10 minutes.
Collagen was of the same origin (Example 1, Experiment 2). An aqueous solution of collagen (2% w/v) was prepared by dissolving 200 mg of collagen in 8.5 ml of an acetic acid solution with a pH about 3.6. Once a clear solution was obtained, it was cooled down to about 4° C., and then a cold solution of 200 mg of BES in 1.5 mL of water was added under continuous stirring. When the resulting neutral solution (pH ˜7.2) appeared quite homogeneous and clear, it was poured in a Petri dish and placed at 37° C. A homogeneous uniform gel formed within 15 minutes.
Collagen (100 mg) of the same origin (Example 1, Experiment 2) was first dissolved in 10 ml of an acetic acid solution (0.1M). Then 100 mg of Chitosan was added to the resulting solution and stirred until all Chitosan was completely dissolved. After the whole system was cooled down to around 4° C., and 200 mg of BES in form of sodium salt dissolved in 1.5 ml of cold water, was added under continuous stirring. Once the solution was perfectly homogeneous and clear, the liquid mixture was poured in a Petri dish and placed at 37° C. The gel formed within 5 minutes.
This example shows the typical experiments of acidic biopolymer solutions neutralized with tertiary hydroxyalkylamine, preferentially bis-[2-hydroxyethyl]iminotris[hydroxymethyl]methane (BIS-TRIS), and transformed to gel upon standing at 37° C.
Chitosan solution with a pH around 5.0 was prepared by dissolving 200 mg of chitosan 85% deacetylated in 8.5 mL of aqueous HCl solution (0.1M). This chitosan solution was cooled down to around 4° C., after which 400 mg of BIS-TRIS dissolved in 1.5 mL of cold water was added slowly to the cold chitosan solution under vigorous stirring. The pH of the resulting homogeneous and clear solution increases to about 7.15. This solution was disposed in a glass scintillation vial in the incubator at 37° C. Bulk gelation occurs within 10 minutes.
While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features hereinbefore set forth, and as follows in the scope of the appended claims.
The present application is a continuation application under 35 U.S.C. §120 of U.S. Ser. No. 10/130,316 filed on Aug. 27, 2007, now abandoned, which is a national stage entry under 35 U.S.C. §317 of International Application No. PCT/CA00/01341 filed on Nov. 10, 2000, which claims benefit of U.S. provisional application Ser. No. 60/165,641 filed on Nov. 15, 1999.
Number | Name | Date | Kind |
---|---|---|---|
2976574 | Keutgen et al. | Mar 1961 | A |
3266906 | Miller et al. | Aug 1966 | A |
3586654 | Lerman et al. | Jun 1971 | A |
3755558 | Scribner | Aug 1973 | A |
3966655 | Kovacs et al. | Jun 1976 | A |
4060081 | Yannas et al. | Nov 1977 | A |
4097935 | Jarcho | Jul 1978 | A |
4185618 | Corey | Jan 1980 | A |
4195175 | Peniston et al. | Mar 1980 | A |
4254207 | Landoll et al. | Mar 1981 | A |
4267313 | Sannan et al. | May 1981 | A |
4337760 | Rubin | Jul 1982 | A |
4391909 | Lim | Jul 1983 | A |
4394373 | Malette et al. | Jul 1983 | A |
4424346 | Hall et al. | Jan 1984 | A |
4454198 | Fickel et al. | Jun 1984 | A |
4474769 | Smith | Oct 1984 | A |
4532134 | Malette et al. | Jul 1985 | A |
4568559 | Nuwayser et al. | Feb 1986 | A |
4605623 | Malette et al. | Aug 1986 | A |
4647536 | Mosbach | Mar 1987 | A |
4659700 | Jackson | Apr 1987 | A |
4678470 | Nashef et al. | Jul 1987 | A |
4722948 | Sanderson | Feb 1988 | A |
4731081 | Tiffany et al. | Mar 1988 | A |
4803075 | Wallace et al. | Feb 1989 | A |
4861627 | Mathiowitz et al. | Aug 1989 | A |
4877775 | Scopelianos | Oct 1989 | A |
4895724 | Cardinal et al. | Jan 1990 | A |
4902792 | Okuma et al. | Feb 1990 | A |
4911926 | Henry et al. | Mar 1990 | A |
4933105 | Fong | Jun 1990 | A |
4956350 | Mosbey | Sep 1990 | A |
4996307 | Itoi et al. | Feb 1991 | A |
5006255 | Uragami | Apr 1991 | A |
5047055 | Bao et al. | Sep 1991 | A |
5049587 | Okamoto et al. | Sep 1991 | A |
5071644 | Viegas et al. | Dec 1991 | A |
5073202 | Wallach et al. | Dec 1991 | A |
5126141 | Henry | Jun 1992 | A |
5204382 | Wallace et al. | Apr 1993 | A |
5266326 | Barry et al. | Nov 1993 | A |
5278201 | Dunn et al. | Jan 1994 | A |
5284833 | McAnalley et al. | Feb 1994 | A |
5294446 | Schlameus et al. | Mar 1994 | A |
5306305 | Lee | Apr 1994 | A |
5318780 | Viegas et al. | Jun 1994 | A |
5368051 | Dunn et al. | Nov 1994 | A |
5414061 | Shimizu et al. | May 1995 | A |
5422116 | Yen et al. | Jun 1995 | A |
5468787 | Braden et al. | Nov 1995 | A |
5489401 | Freeman | Feb 1996 | A |
5587175 | Viegas et al. | Dec 1996 | A |
5599552 | Dunn et al. | Feb 1997 | A |
5612028 | Sackier et al. | Mar 1997 | A |
5618339 | Ito | Apr 1997 | A |
5620706 | Dumitriu et al. | Apr 1997 | A |
5626861 | Laurencin et al. | May 1997 | A |
5655546 | Halpern | Aug 1997 | A |
5658593 | Orly et al. | Aug 1997 | A |
5683461 | Lee et al. | Nov 1997 | A |
5709854 | Griffith-Cima et al. | Jan 1998 | A |
5723331 | Tubo et al. | Mar 1998 | A |
5736372 | Vacanti et al. | Apr 1998 | A |
5749874 | Schwartz | May 1998 | A |
5770193 | Vacanti et al. | Jun 1998 | A |
5770417 | Vacanti et al. | Jun 1998 | A |
5773033 | Cochrum et al. | Jun 1998 | A |
5773608 | Yen | Jun 1998 | A |
5811094 | Caplan et al. | Sep 1998 | A |
5820608 | Luzio et al. | Oct 1998 | A |
5830503 | Chen | Nov 1998 | A |
5837235 | Mueller et al. | Nov 1998 | A |
5842477 | Naughton et al. | Dec 1998 | A |
5843156 | Slepian et al. | Dec 1998 | A |
5855619 | Caplan et al. | Jan 1999 | A |
5866415 | Villeneuve | Feb 1999 | A |
5871985 | Aebischer | Feb 1999 | A |
5874500 | Rhee et al. | Feb 1999 | A |
5894070 | Hansson et al. | Apr 1999 | A |
5900238 | Gombotz et al. | May 1999 | A |
5902741 | Purchio et al. | May 1999 | A |
5902798 | Gouda et al. | May 1999 | A |
5906934 | Grande et al. | May 1999 | A |
5908784 | Johnstone et al. | Jun 1999 | A |
5942487 | Ogawa et al. | Aug 1999 | A |
5944754 | Vacanti | Aug 1999 | A |
5958443 | Viegas et al. | Sep 1999 | A |
5964807 | Gan et al. | Oct 1999 | A |
5977330 | Lohmann et al. | Nov 1999 | A |
5977930 | Fischer et al. | Nov 1999 | A |
5989579 | Darougar et al. | Nov 1999 | A |
6005161 | Brekke et al. | Dec 1999 | A |
6060534 | Ronan et al. | May 2000 | A |
6080194 | Pachence et al. | Jun 2000 | A |
6110209 | Stone | Aug 2000 | A |
6124273 | Drohan et al. | Sep 2000 | A |
6136334 | Viegas et al. | Oct 2000 | A |
6179872 | Bell et al. | Jan 2001 | B1 |
6200606 | Peterson et al. | Mar 2001 | B1 |
6344488 | Chenite et al. | Feb 2002 | B1 |
6372257 | Marchosky | Apr 2002 | B1 |
6417247 | Armonstrong et al. | Jul 2002 | B1 |
6425918 | Shapiro et al. | Jul 2002 | B1 |
6482223 | Nowakowski et al. | Nov 2002 | B1 |
6610669 | Calias et al. | Aug 2003 | B1 |
6632468 | Morgan et al. | Oct 2003 | B2 |
6649192 | Alonso Fernandez et al. | Nov 2003 | B2 |
6706690 | Reich et al. | Mar 2004 | B2 |
6743783 | Vournakis et al. | Jun 2004 | B1 |
6756363 | Nordquist et al. | Jun 2004 | B1 |
6911212 | Gertzman et al. | Jun 2005 | B2 |
7045141 | Merboth et al. | May 2006 | B2 |
7148209 | Hoemann et al. | Dec 2006 | B2 |
7320962 | Reich et al. | Jan 2008 | B2 |
7368126 | Chen et al. | May 2008 | B2 |
7459307 | Ha et al. | Dec 2008 | B2 |
8258117 | Hoemann et al. | Sep 2012 | B2 |
20020068048 | Dreyfus et al. | Jun 2002 | A1 |
20020082220 | Hoemann et al. | Jun 2002 | A1 |
20030143274 | Viegas et al. | Jul 2003 | A1 |
20030147860 | Marchosky et al. | Aug 2003 | A1 |
20030158302 | Chaput et al. | Aug 2003 | A1 |
20030199615 | Chaput et al. | Oct 2003 | A1 |
20040013733 | Chen et al. | Jan 2004 | A1 |
20040022859 | Chen et al. | Feb 2004 | A1 |
20040024069 | Chen et al. | Feb 2004 | A1 |
20040047892 | Desrosiers et al. | Mar 2004 | A1 |
20040091540 | Desrosiers et al. | May 2004 | A1 |
20050244393 | Phillipart et al. | Nov 2005 | A1 |
20060004189 | Gandy | Jan 2006 | A1 |
20060008524 | Chen et al. | Jan 2006 | A1 |
20060018973 | Kim et al. | Jan 2006 | A1 |
20060062768 | Hnojewyj | Mar 2006 | A1 |
20060193892 | Furst et al. | Aug 2006 | A1 |
20060204544 | Sunwoo et al. | Sep 2006 | A1 |
20060204581 | Gower et al. | Sep 2006 | A1 |
20060263401 | Rubsamen | Nov 2006 | A1 |
20060293216 | Klaveness et al. | Dec 2006 | A1 |
20070014867 | Kusanagi et al. | Jan 2007 | A1 |
20070037737 | Hoemann et al. | Feb 2007 | A1 |
20070167400 | Boucher et al. | Jul 2007 | A1 |
20070254007 | Bumgardner et al. | Nov 2007 | A1 |
20080118563 | Muzzarelli et al. | May 2008 | A1 |
20080200430 | Bitterman et al. | Aug 2008 | A1 |
20080248991 | Dyer et al. | Oct 2008 | A1 |
20090004276 | Ben-Shalom et al. | Jan 2009 | A1 |
Number | Date | Country |
---|---|---|
2219399 | Apr 1999 | CA |
2319558 | Sep 1999 | CA |
2329329 | Oct 1999 | CA |
2412605 | Nov 2001 | CA |
0 298 501 | Jan 1980 | EP |
0298501 | Jan 1980 | EP |
0539751 | Jan 1992 | EP |
94112944 | Aug 1994 | EP |
94112944 | Jan 1995 | EP |
0543572 | Jul 1997 | EP |
0884052 | Dec 1998 | EP |
1 077 253 | Feb 2001 | EP |
1 077 253 | Feb 2001 | EP |
0 640 647 | Mar 2001 | EP |
2 246 514 | Feb 1992 | GB |
2 277 915 | Nov 1994 | GB |
WO 9325191 | Dec 1993 | WO |
WO 9400134 | Jan 1994 | WO |
WO 9525549 | Sep 1995 | WO |
WO 9623039 | Jan 1996 | WO |
WO 9602276 | Feb 1996 | WO |
WO 9639202 | Dec 1996 | WO |
WO 9733562 | Sep 1997 | WO |
WO 9741899 | Nov 1997 | WO |
WO 9822114 | May 1998 | WO |
WO 9904720 | Feb 1999 | WO |
WO 9907416 | Feb 1999 | WO |
WO 9922747 | May 1999 | WO |
WO 9947130 | Sep 1999 | WO |
WO 9947186 | Sep 1999 | WO |
WO 0002905 | Jan 2000 | WO |
WO 0044413 | Aug 2000 | WO |
WO 0048550 | Aug 2000 | WO |
0136000 | May 2001 | WO |
WO 0136000 | May 2001 | WO |
WO 0136000 | May 2001 | WO |
WO 0141822 | Jun 2001 | WO |
WO 0141822 | Jun 2001 | WO |
0200272 | Jan 2002 | WO |
WO 0200272 | Jan 2002 | WO |
WO 0240070 | May 2002 | WO |
WO 0240070 | May 2002 | WO |
WO 03042250 | May 2003 | WO |
2004016297 | Feb 2004 | WO |
WO 2004016297 | Feb 2004 | WO |
2008064487 | Jun 2008 | WO |
WO 2008064487 | Jun 2008 | WO |
Entry |
---|
Gelrite 082006. |
Gelrite 2007. |
Aerts et al., Journal of Biomechanics, 28(11):1299-1308 (1995). |
Aiba, Makromol. Chemie, 194(1):65-75 (1993). |
Alexander et al., Journal of Zoology—London (A), 209:405-419 (1986). |
Appling et al., FEBS Letters, 250(2):541-544 (1989). |
Aston et al., Journal of Bone and Joint Surgery, 68-B(1):29-35 (1986). |
Ateshian, Journal of Biomechanical Engineering, 119:81-86 (1997). |
Austin et al., Science, 212:749-753 (1981). |
Balkin, Neale's Common Foot Disorders: Diagnosis and Management, 22:387-400 (1997). |
Bartone et al., Journal of Urology, 140:1134-1137 (1988). |
Bennett et al., Journal of Anatomy, 171:131-138 (1990). |
Bentley et al., Nature, 230:385-388 (1971). |
Bernkop-Schnurch et al., Journal of Pharmaceutical Sciences. 87(4):430-434 (1998). |
Blechschmidt, Foot and Ankle, 2(5):260-283 (1982). |
Bobic et al., The Journal of Bone and Joint Surgery, 82-B(2):165-166 (2000). |
Breinan et al., The Journal of Bone and Joint Surgery, 79-A(10):1439-1451 (1997). |
Breinan et al., Journal of Orthopaedic Research, 18(5):781-789 (2000). |
Brittberg et al., The New England Journal of Medicine, 331(14):889-895 (1994). |
Brittberg et al., Clinical Orthopaedics and Related Research, 326:270-283 (1996). |
Buckwalter et al., The Journal of Bone and Joint Surgery, 79-A(4):612-632 (1997). |
Buschmann et al., Journal of Orthopaedic Research, 10(6):745-758 (1992). |
Buschmann et al., Foot and Ankle, 14(7):389-394 (1993). |
Buschmann et al., Foot and Ankle, 16(5):254-258 (1995). |
Butnariu-Ephrat et al., Clinical Orthopaedics and Related Research, 330:234-243 (1996). |
Caplan et al., Clinical Orthopaedics and Related Research, 342:254-269 (1997). |
Carreño-Gómez et al., International Journal of Pharmaceutics, 148:231-240 (1997). |
Chenite et al., Carbohydrate Polymers, 00:1-9 (2000). |
Chesterman et al., The Journal of Bone and Joint Surgery, 50B(1):184-197 (1968). |
Childers et al., Clinical Orthopaedics and Related Research, 144:114-120 (1979). |
Cho et al., Biomaterials, 20:2139-2145 (1999). |
Chu et al., Journal of Biomedical Materials Research, 29:1147-1154 (1995). |
Chu et al., Clinical Orthopaedics and Related Research, 340:220-229 (1997). |
Cohen et al., British Journal of Haemotology, 31:45-50 (1975). |
D'Ambrosia, Orthopedics, 10(1):137-142 (1987). |
Denuziere et al., Biomaterials, 19:1275-1285 (1998). |
DePalma et al., Clinical Orthopaedics and Related Research, 48:229-242 (1966). |
Dillon et al., J. Biomater. Sci. Polymer Edn., 9(10):1049-1069 (1998). |
Elçin et al., Neurological Research, 20:648-654 (1998). |
Frenkel et al., The Journal of Bone and Joint Surgery, 79-B(5):831-836 (1997). |
Freed et al., Journal of Biomedical Materials Research, 28:891-899 (1994). |
Fukamizo et al., Biochem. Cell Biol., 75:687-696 (1997). |
Gillquist et al., Acta Orthop Scand., 68(2):186-191 (1997). |
Grande et al., Journal of Orthopaedic Research, 7(2):208-218 (1989). |
Green, Clinical Orthopaedics and Related Research, 124:237-250 (1977). |
Hangody et al., Knee Surg., Sports Traumatol., Arthrosc., 5:262-267 (1997). |
Hangody et al., Foot and Ankle International, 18(10):628-634 (1997). |
Hendrickson et al., Journal of Orthopaedic Research, 12(4):485-497 (1994). |
Higaki et al., JSME International Journal, 40(4):776-781 (1997). |
Hirano et al., Biopolymers, 15:1685-1691 (1976). |
Homminga et al., Acta Orthop. Scand., 62(5):415-418 (1991). |
Hunziker et al., The Journal of Bone and Joint Surgery, 78-A(5):721-733 (1996). |
Hyc et al., Cell Transplantation, 6(2):119-124 (1997). |
Itay et al., Cartilage Repair by Cultured Chondrocytes, 220:284-303 (1987). |
Jahss et al., Foot and Ankle, 13(5):227-232 (1992). |
Johnson, Operative Arthroscopy, Chapter 24, pp. 341-360 (1991). |
Jürgensen et al., The Journal of Bone and Joint Surgery, 79-A(2):185-193 (1997). |
Kandel et al., Art. Cells, Blood Subs., and Immob. Biotech., 23(5):565-577 (1995). |
Kawamura et al., Acta Orthop. Scand., 69(1):56-62 (1998). |
Ker, Journal of Experimental Biology, 199:1501-1508 (1996). |
Kopp et al., Int. J. Cancer, 60:275-279 (1995). |
Koyano et al., J. Biomed. Mater. Res., 39:486-490 (1998). |
Kubota et al., Polymer Journal, 29(2):123-127 (1997). |
Kuettner, Clinical Biochemistry, 25:155-163 (1992). |
Lahiji et al., J. Biomed. Mater. Res., 51:586-595 (2000). |
Lee et al., Journal of Controlled Release, 51:213-220 (1998). |
Lee et al., J. Periodontol., 71(3):410-417 (2000). |
Leistikow, Seminars in Thrombosis and Hemostasis, 22(3):289-294 (1996). |
Lu et al., Biomaterials, 20:1937-1944 (1999). |
Mahomed et al., Orthopedics, 15(10):1191-1199 (1992). |
Malette et al., The Annals of Thoracic Surgery, 36(1):55-58 (1983). |
Mankin, The New England Journal of Medicine, pp. 1285-1292 (1974). |
Mattioli-Belmonte et al., Medical and Biological Engineering and Computing, 37:130-134 (1999). |
Messner et al., Acta Orthop. Scand., 67(5):523-529 (1996). |
Minas et al., Articular Cartilage Defects, 20(6):525-538 (1997). |
Muzzarelli et al., Eur. Chitin Soc., Ancona (1993). |
Muzzarelli et al., Enzyme Microb. Technol., 17:541-545 (1995). |
Namba et al., The Journal of Bone and Joint Surgery, 80-A(1):4-10 (1998). |
Narváez et al., Radiographics, 20(2):333-352 (2000). |
Nevo et al., Cell Transplantation, 7(1):63-70 (1998). |
Newman, The American Journal of Sports Medicine, 26(2):309-324 (1998). |
Nixon et al., Journal of Orthopaedic Research, 17(4):475-487 (1999). |
Noguchi et al., Clinical Orthopaedics and Related Research, 302:251-258 (1994). |
O'Driscoll et al., The Journal of Bone and Joint Surgery, 70-A(4):595-606 (1988). |
O'Driscoll et al., The Journal of Bone and Joint Surgery, 76-A(7):1042-1051 (1994). |
Okamoto et al., J. Vet. Med. Sci., 57(5):851-854 (1995). |
Outerbridge et al., The Journal of Bone and Joint Surgery, 77-A(1):65-72 (1995). |
Paletta et al., The American Journal of Sports Medicine, 20(6):725-731 (1992). |
Pechak et al., Bone, 7:459-472 (1986). |
Peluso et al., Biomaterials, 15(15):1215-1220 (1994). |
Pridie, The Journal of Bone and Joint Surgery, 41-B(3):618-619 (1959). |
Robinson et al., Calcif Tissue Int., 46:246-253 (1990). |
Rodrigo et al., Operative Orthopaedics, Chapter 144, pp. 2077-2082 (1993). |
Sall et al., Ann. Ophthalmol., 19:31-33 (1987). |
Sams et al., Osteoarthritis and Cartilage, 3:47-59 (1995). |
Schipper et al., Pharmaceutical Research, 14(7):923-929 (1997). |
Schwarz et al., British Journal of Rheumatology, 37(1):21-26 (1998). |
Sechriest et al., J. Biomed. Mater Res, 49(4):534-541 (2000). |
Sellers et al., The Journal of Bone and Joint Surgery, 79-A(10):1452-1463 (1997). |
Sellers et al., The Journal of Bone and Joint Surgery, 82-A(2):151-160 (2000). |
Shephard et al., XVIIth FECTS Meeting Patras, Greece, Abstract Form (Jul. 1-5, 2000). |
Shigemasa et al., Biotechnology and Genetic Engineering Reviews, 13:383-420 (1995). |
Soulhat et al., Journal of Biomechanical Engineering, 121:340-347 (1999). |
Specchia et al., Bulletin for Hospital for Joint Diseases, 54(4):230-235 (1996). |
Steadman et al., J. Sports Traumatol. Rel. Res., 20(2):61-70 (1998). |
Stone et al., British Journal of Plastic Surgery, 53:601-606 (2000). |
Suh et al., Biomaterials, 21:2589-2597 (2000). |
Terbojevich et al., Carbohydrate Polymers, 29(1):63-68 (1996). |
Ueno et al., Biomaterials, 20:1407-1414 (1999). |
Van Schie et al., Diabetes Care, 23(5):634-638 (2000). |
Vasios et al., 45th Annual Meeting, Orthopaedic Research Society, Anaheim, California, 711 (Feb. 1-4, 1999). |
Wakitani et al., The Journal of Bone and Joint Surgery, 71-B(1):74-80 (1989). |
Wakitani et al., The Journal of Bone and Joint Surgery, 76-A(4):579-592 (1994). |
Wei et al., Journal of Biomedical Materials Research, 34:63-72 (1997). |
Yagi et al., Biol. Pharm. Bull., 20(12):1290-1294 (1997). |
Zoppou et al., Bulletin of Mathematical Biology, 59(5):953-973 (1997). |
Chenite, A. et al., “Novel Injectable Neutral Solutions of Chitosan Form Biodegradable Gels In Situ.” Biomaterials 21(21):2155-2161, 2000. |
Ohya Y. et al. J. Microencapsulation, 10(1):1-9, 1993. |
Zielinski B.A. et al. Biomaterials, 15(13):1049-1056, 1994. |
Chung C.H. et al., Calcif Tissue Int., 51:305-311, 1992. |
Bellows C.G. et al., Bone and Mineral, 17:15-29, 1992. |
Guo J. et al., Connective Tissue Research, 19:277-297, 1989. |
Li, X., Biotechnol. Appl. Biochem., 23:269-271, 1996. |
Gupta S. et al., The International Journal of Artificial Organs, 16(3):155-163, 1993. |
Matthew H.W.T. et al., Journal of Pediatric Surgery, 28(11):1423-1428, 1993. |
Rao, S. Bhaskara et al., Journal of Biomedial Materials Research, 34:21-28, 1997. |
Muzzarelli R. et al., Biomaterials, 9:247-252, 1988. |
Muzzarelli R.A.A. et al., Biomaterials, 15(13):1075-1081, 1994. |
Calvo P. et al., Colloid Polym, Sci., 275:46-53, 1997. |
Aspden T.J. et al., European Journal of Pharmaceutical Sciences, 4:23-31, 1996. |
Abstract, AN 1990:25365, Senoo et al., 1990. |
Peluso et al., Biomaterials, 15(15):1215-1220, 1994. |
Pridie, The Journal of Bone and Joint Surgery, 41-B(3):618-619, 1959. |
Rao et al., Journal of Biomedical Materials Research, 34:21-28, 1997. |
Robinson et al., Calcif Tissue Int., 46:246-253, 1990. |
Rodrigo et al., Operative Orthopaedics, 2077-2082, 1993. |
Sall et al., Ann Ophtalmol., 19:31-33, 1987. |
Sams et al., Osteoarthritis and Cartilage, 3:47-59, 1995. |
Schipper et al., Pharmaceutical Research, 14(7):923-929, 1997. |
Schwarz et al., British Journal of Rheumatology, 37(1):21-26, 1998. |
Sechriest et al., J. Biomed. Mater. Res., 49(4):534-541, 2000. |
Sellers et al., The Journal of Bone and Joint Surgery, 79-A:1452-1463, 1997. |
Sellers et al., The Journal of Bone and Joint Surgery, 82-A(2):151-160, 2000. |
Shephard et al., XVIIth FECTS Meeting Patras, Greece, Abstract Form, Jul. 1-5, 2000. |
Shigemasa et al., Biotechnology and Genetic Engineering Reviews, 13:383-420, 1995. |
Soulhat et al., Journal of Biomechanical Engineering, 121:340-347, 1999. |
Specchia et al., Bulletin for Hospital for Joint Diseases, 54(4):230-235, 1996. |
Steadman et al., J. Sports Traumatol. rel. res., 20(2):61-70, 1998. |
Stone et al., British Journal of Plastic Surgery, 53:601-606, 2000. |
Suh et al., Biomaterials, 21:2589-2598, 2000. |
Terbojevich et al., Carbohydrate Polymers, 29(1):63-68, 1996. |
Ueno et al., Biomaterials, 20:1407-1414, 1999. |
Van Schie et al., Diabetes Care, 23(5):634-638, 2000. |
Vasios et al., 45th Annual Meeting, Orthopaedic Research Society, Anaheim, California, 711, Feb. 1-4, 1999. |
Wakitani et al., The Journal of Bone and Joint Surgery, 71-B(1):74-80, 1989. |
Wakitani et al., The Journal of Bone and Joint Surgery, 76-A(4):579-592, 1994. |
Wei et al, Journal of Biomedical Materials Research, 34:63-72, 1997. |
Yagi et al, Biol. Pharm. Bull, 20(12):1290-1294, 1997. |
Zielinski et al. Biomaterials, 15(13): 1049-1056, 1994. |
Zoppou et al., Bulletin of Mathematical Biology, 59(5):953-973, 1997. |
Hawley's Condensed Chemical Dictionary, 1993, pp. 256. |
Mosbach, 1988, Methods Enzymol., 137: 443. |
Arshady R., 1993, Biomaterials, 14: 5. |
Jalil R. and Nixon J.R., 1990, J. Microencapsul., 7: 297. |
Bodmeier R. and McGinity J.W., 1987, J. Microencapsul., 7: 279. |
Gillquist et al., 1997, Acta Orthop Scand., 68(2): 186-191. |
Chen et al., 2003, Langmuir, 19: 9382-9386. |
Shimizu et al., (Nippon Kagaku Kaishi (1998), (9), 637-641) (Abstract Sent). |
Ruel-Gariepy et al., 2000, International Journal of Pharmaceutics, 203: 89-98. |
Arnoczky, S.P., R.F. Warren and J. M. Spivak. 1988. “Meniscal repair using an exogenous fibrin clot. An experimental study in dogs.” J. Bone Joint Sug Am 70, No. 8, p. 1209-17. |
Clark, Richard A.F. 1996. The molecular and cellular biology of wound repair. 2 ed. New York: Plenum. |
Hall, B.K. 1983. Cartilage. New York, NY: Academic Press. |
Insall, J.N. 1967. Intra-articular surgery for degenerative arthritis of the knee. A report of the work of the late K.H. Pridie. J. Bone Joint Surg Br 49, No. 2, p. 211-28. |
Inui, H., M. Tsujikubo, S. Hirano. 1995. Biosci Biotechnol Biochem, 59: 211-214. |
McCarthy, D.J. and W.J. Koopman. 1993. “Arthritis and allied conditions. A textbook of rheumatology.” Philadelphia: Lea and Febiger. |
Sashiwa, H., H. Saimoto, Y. Shigemasa, R. Ogawa and S. Tokura. 1990. International Journal of Biological Macromolecules, 12: 295-296. |
Yalpani, M and D. Pantaleone. 1994. “An examination of the unusual suceptbilities of aminoglycans to enzymatic hydrolysis.” Carbohydrate Research 256, No. 1, p. 159-75. |
Hsien, T.Y. and Rorrer G.L., 1997, Ind. Eng. Chem. Res., 36: 3631-3638. |
Lavertu et al., 2003, J. Pharmaceutical Biomedical Analysis, 32: 1149-1158. |
Liu et al., 2003, Bioconjugate Chem., 14: 782-789. |
Muzzarelli et al., 2001, Biomacromolecules, 2: 165-169. |
Capitani et al., 2001, Carbohydrate Polymers, 45: 245-252. |
De Angelis, 1998, Macromolecules, 31: 1595-1601. |
Kumbar et al., 2002, J. Microencapsulation, 19: 173-180. |
Li et al., 2002, Journal of Pharmaceutical Sciences, 91: 1669-1677. |
Mi et al., 2002, Biomaterials, 23: 181-191. |
Rogovina et al., 2001, Polymer Science, 43: 265-268. |
Suto et al., 1996, Journal of Applied Polymer Science, 61: 2273-2218. |
Colman et al., 2001, Chapter 1, Hemostasis and Thrombosis, Basic Principles & Clinical Practice, Lippincott Williams & Wilkins, 4th Ed. |
Fan et al., 2005, J Thrombosis and Hemostasis, 3: 1056-1063. |
Buschmann et al., 2006, Cartilage repair with chitosan/glycerol-phosphate stabilised blood clots, Edited by Riley J Williams Humana Press. |
Rivard et al., 2005, J Thromb Haemost, 3: 2039-2043. |
Jamieson et al., 1924, J. Am. Chem. Soc., 46: 775-778. |
Hoemann et al., 2005, J Bone Joint Surg Am, 87: 2671-2686. |
Hoemann et al., 2007, Osteoarthritis Cartilage, 15: 78-89. |
Chevrier et al., 2007, Osteoarthritis Cartilage, 15: 316-327. |
English translation of JP 10-259134 A, Niimura et al., 1998. |
Gooding et al., 1986, Investigative Radiology, 21: 45-48. |
Resnick et al., 1999, Foot & Ankle International, 20: 481-484. |
Ruel-Gariepy et al., 2004, European Journal of Pharmaceutics and Biopharmaceutics, 58: 409-426. |
Determan et al., Polymer, 46: 6933-6946, 2005. |
Ghzaoui et al., Langmuir, 2004, 17: 9348-9353. |
English translation of Japanese Patent Application No. 2002-506814. |
Hoemann et al., 2007, J Biomed Materials Res Part A, 83A: 521-529. |
Eroglu et al., 2002, International Journal of Pharmaceutics, 235: 51-59. |
Dodane et al., 1998, Pharmaceutical Science and Technology Today, 1: 246-253. |
Ruel-Gariepy et al., 2006, ACS Symposium Series Amer Chemical Soc, 1155 Sixteenth ST NW, Washington, DC 20036 USA Series : ACS Symposium Series , pp. 243-224. |
Marchand et al., 2009, Osteoarthritis and Cartilage , 17: 953-960. |
Hangody et al., Knee Surg. Sports Traumatol, Arthrosc., 5:262-267, 1997. |
Hangody et al., Foot & Ankle International, 18(10):628-634, 1997. |
Hendrickson et al., Journal of Orthopaedic Research, 12(4):485-497, 1994. |
Higaki et al., JSME International Journal, 40(4):776-781, 1997. |
Hirano et al., Biopolymers, 15:1685-1691, 1976. |
Homminga et al., Acta Orthop. Scand., 62(5):415-418, 1991. |
Hunziker et al., The Journal of Bone and Joint Surgery, 78-A(5):721-733, 1996. |
Hyc et al., Cell Transplantation, 6(2):119-124, 1997. |
Itay et al., Cartilage Repair by Cultured Chondrocytes, 220:284-303, 1987. |
Jahss et al., Foot & Ankle, 13(5):227-232, 1992. |
Johnson, Operative Arthroscopy, Chapter 24, 341-360, 1991. |
Jürgensen et al., The Journal of Bone and Joint Surgery, 79-A(2):185-193, 1997. |
Kandel et al., Art. Cells, Blood Subs., and Immob. Blofech., 23(5):565-577, 1995. |
Kawamura et al., Acta Orthop. Scand., 69(1):56-62, 1998. |
Ker, Journal of Experimental Biology, 199:1501-1508, 1996. |
Kopp et al., Int. J. Cancer, 60:275-279, 1995. |
Koyano et al., J. Biomed. Mater. Res., 39:486-490, 1998. |
Kubota et al., Polymer Journal, 29(2):123-127, 1997. |
Kuettner, Clinical Biochemistry, 25:155-163, 1992. |
Lahiji et al., Matrix Proteins inHuman Osteoblasts, 586-595, 2000. |
Lee et al., Journal of controlled release, 51:213-220, 1998. |
Lee et al., J. Periodontol., 71(3):410-417, 2000. |
Leistikow, Seminars in Thrombosis and Homostasis, 22(3):289-294, 1996. |
Li, Biotechnol. Appl. Biochem., 23:269-271, 1996. |
Lu et al., Biomaterials, 20:1937-1944, 1999. |
Mahomed et al., Orthopedics, 15(10):1191-1199, 1992. |
Sei-Ichi Aiba, Makromolekulare Chemie, 194(1), 65-75, 1993. |
Malette et al., The Annals of Thoracic Surgery, 36(1):55-58, 1983. |
Mankin, The New England Journal of Medicine, 1285-1293, 1974. |
Matthew et al., Journal of Pediatric Surgery, 28(11):1423-1428, 1993. |
Mattioli-Belmonte et al., Medical & Biological Engineering & Computing, 37:130-134, 1999. |
Messner et al., Acta Orthop. Scand., 67(5):523-529, 1996. |
Minas et al., Articular Cartilage Defects, 20(6):525-538, 1997. |
Muzzarelli et al., Biomaterials, 9:247-252, 1988. |
Muzzarelli et al., Eur. Chitin Soc., Ancona, 1993. |
Muzzarelli et al., Biomaterials, 15(13):1075-1081, 1994. |
Muzzarelli et al., Enzyme Microb. Technol., 17:541-545, 1995. |
Namba et al., The Journal of Bone and Joint Surgery, 80-A(1):4-10, 1998. |
Narvaez et al., Radiographics, 20(2):333-352, 2000. |
Nevo et al., Cell Transplantation, 7(1 ):63-70, 1998. |
Newman, The American Journal of Sports Medicine, 26(2):309-324, 1998. |
Nixon et al., Journal of Orthopaedic Research, 17(4):475-487, 1999. |
Noguchi et al., Clinical Orthopaedics and Related Research, 302:251-258, 1994. |
O'Driscoll et al., The Journal of Bone and Joint Surgery, 70-A(4):595-606, 1988. |
O'Driscoll et al, The Journal of Bone and Joint Surgery, 76-A(7):1042-1051, 1994. |
Ohya et al. J. Microencapsulation, 10(1):1-9, 1993. |
Okamoto et al., J V et Med. Sci., 57(5): 851-854, 1995. |
Outerbridge et al., The Journal of Bone and Joint Surgery, 77-A(1):65-72, 1995. |
Paletta et al., The American Journal of Sports Medicine, 20(6):725-731, 1992. |
Pechak et al., Bone, 7:459-472, 1986. |
Aerts et al., Journal of Biomechanics, 28:1299-1308, 1995. |
Alexander et al., Journal of Zoology—London (A):209:405-419, 1986. |
Appling et al., FEBS Letters, 250(2):541-544, 1989. |
Aspden et al., European Journal of Pharmaceutical Sciences, 4:23-31, 1996. |
Aston et al., The Journal of Bone and Joint Surgery, 68-B(1):29-35, 1986. |
Ateshian, Journal of Biomechanical Engineering, 119:81-86, 1997. |
Austin et al., Science, 212:749-753, May 15, 1981. |
Back et al., Biochemistry, 18(23): 5191-5196, 1979. |
Balkin in Neale's common foot disorders: diagnosis and management, 22:387-400, 1997. |
Bartone et al., The Journal of Urology, 140:1134-1137, 1988. |
Bellows et al., Bone and Mineral, 17:15-29, 1992. |
Bennett et al., Journal of Anatomy, 171:131-138, 1990. |
Bentley et al., Nature, 230:385-388, 1971. |
Bernkop-Schnurch et al., Journal of Pharmaceutical Sciences, 87 (4):430-434, 1998. |
Blechschmidt, Foot & Ankle, 2(5):260-283, 1982. |
Bobic et al., The Journal of Bone and Joint Surgery, 82-B(2):165-166, 2000. |
Breinan et al., The Journal of Bone and Joint Surgery, 79-A(10):1439-1451, 1997. |
Breinan et al., Journal of Orthopaedic Research, 18(5):781-789, 2000. |
Brittberg et al., Acta Orthop Scand., 68(2):186-191, 1997. |
Brittberg et al., The New England Journal of Medicine, 331(14):889-895, 1994. |
Brittberg et al., Clinical Orthopaedics and Related Research, 326:270-283, 1996. |
Buckwalter et al., The Journal of Bone and Joint Surgery, 79-A(4):612-632, 1997. |
Buschmann et al., Journal of Orthopaedic Research, 10(6):745-758, 1992. |
Buschmann et al., Foot & Ankle, 14:389-394, 1993. |
Buschmann et al., Foot & Ankle, 16:254-258, 1995. |
Butnariu-Ephrat et al., Clinical Orthopaedics and Related Research, 330:234-243, 1996. |
Calvo et al., Colloid & Polymer Science, (Abstract) 275(1):46-53, 1997. |
Caplan et al., Clinical Orthopaedics and Related Research, 342:254-269, 1997. |
Carreno-Gomez et al., International Journal of Pharmaceutics, 148:231-240, 1997. |
Chenite et al., Carbohydrate Polymers, 46(1):39-47, 2001. |
Chenite, et al., Biomaterials, 21:2155-2161, 2000. |
Chesterman et al., The Journal of Bone and Joint Surgery, 50B(1):184-197, 1968. |
Childers et al., Clinical Orthopaedics and Related Research, 144:114-120, 1979. |
Cho et al., Biomaterials, 20:2139-2145, 1999. |
Chu et al, Journal of Biomedical Materials Research, 29:1147-1154, 1995. |
Chu et al., Clinical Orthopaedics and Related Research, 340:220-229, 1997. |
Chung et al., Calcif Tissue Int., 51:305-311, 1992. |
Cohen et al., British Journal of Haemotology, 31:45-50, 1975. |
D'Ambrosia, Orthopedics, 10(1):137-142, 1987. |
Denuzière et al., Biomaterials, 19:1275-1285, 1998. |
DePalma et al, Clinical Orthopaedics and Related Research, 48:229-242, 1966. |
Dillon et al., J. Biomater Sci. Polymer Edn., 9(10):1049-1069, 1998. |
Elçin et al., Neurological Research, 20:648-654, 1998. |
Frenkel et al., The Journal of Bone and Joint Surgery, 79-B(5): 831-836, 1997. |
Freed et al., Journal of Biomedical Materials Research, 28:891-899, 1994. |
Fukamizo et al., Biochem. Cell Biol., 75:687-696, 1997. |
Grande et al., Journal of Orthopaedic Research, 7(2):208-218, 1989. |
Green, Clinical Orthopaedics and Related Research, 124:237-250, 1977. |
Guo et al., Connective Tissue Research, 19:277-297, 1989. |
Gupta et al., The International Journal of Artificial Organs, 16(3):155-163, 1993. |
Number | Date | Country | |
---|---|---|---|
20100028434 A1 | Feb 2010 | US |
Number | Date | Country | |
---|---|---|---|
60165641 | Nov 1999 | US |
Number | Date | Country | |
---|---|---|---|
Parent | 10130316 | US | |
Child | 12576354 | US |