Patent Application
20210213181
The present application relates to shear thinning compositions which are useful for reducing and/or preventing tissue adhesion in a subject (e.g., a post-operative subject).
More than 90% of surgical patients develop postoperative adhesions, and the incidence of hospital re-admission due to complications related to this pathology can be as high as 20%. Current adhesion barriers present limited efficacy and low clinical adoption due to difficulties in application on irregular surfaces, and incompatibility with minimally invasive interventions.
The present application provides, inter alia, a composition, comprising:
about 3% to about 15% by weight laponite; and
about 0.5% to about 10% by weight poly(ethylene oxide).
In some embodiments, the composition comprises about 3% to about 10% laponite. In some embodiments, the composition comprises about 5% to about 10% laponite. In some embodiments, the composition comprises about 8% to about 10% laponite. In some embodiments, the laponite comprises a positively charged edge and a negatively charged surface. In some embodiments, the overall charge of the laponite is negative.
In some embodiments, the laponite has an average diameter of from about 5 nm to about 60 nm. In some embodiments, the laponite has an average diameter of from about 10 nm to about 40 nm. In some embodiments, the laponite has an average diameter of from about 20 nm to about 30 nm.
In some embodiments, the laponite has an average thickness of from about 0.5 nm to about 2 nm. In some embodiments, the laponite has an average thickness of about 1 nm.
In some embodiments, the composition comprises about 1% to about 3% by weight poly(ethylene oxide). In some embodiments, the composition comprises about 3% to about 5% by weight poly(ethylene oxide). In some embodiments, the poly(ethylene oxide) has an average molecular weight of about 18,000 to about 22,000 g/mol.
In some embodiments, the composition comprises:
about 8% to about 10% by weight laponite and about 1% to about 5% by weight poly(ethylene oxide); or
about 5% to about 10% by weight laponite and about 1% to about 3% by weight poly(ethylene oxide); or
about 8% to about 10% by weight laponite and about 1% to about 3% by weight poly(ethylene oxide).
In some embodiments, the composition comprises:
about 5% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 5% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 5% by weight laponite and about 3% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 3% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 3% by weight poly(ethylene oxide).
In some embodiments, the composition further comprises water. In some embodiments, the composition comprises about 85% to about 94% by weight water.
In some embodiments, the composition comprises:
about 8% to about 10% by weight laponite, about 1% to about 5% by weight poly(ethylene oxide), and about 85% to about 91% by weight water; or
about 5% to about 10% by weight laponite, about 1% to about 3% by weight poly(ethylene oxide), and about 87% to about 94% by weight water; or
about 8% to about 10% by weight laponite, about 1% to about 3% by weight poly(ethylene oxide), and about 87% to about 91% by weight water.
In some embodiments, the composition consists of laponite, poly(ethylene oxide), and water.
In some embodiments, the water is deionized water.
In some embodiments, the composition is a gel. In some embodiments, the composition is a hydrogel.
In some embodiments, the laponite consists of about 66% SiO2, about 30% MgO, about 3% Na2O, and about 1% LiO2.
The present application further provides a composition comprising about 5% to about 10% by weight laponite, about 1% to about 5% by weight poly(ethylene oxide), and about 85% to about 94% by weight water, wherein the composition is prepared according to a process comprising:
(a) combining the laponite and water to form a first mixture;
(b) adding the poly(ethylene oxide) to the first mixture to form the composition.
In some embodiments, the yield stress of the composition is from about 100 Pa to about 2000 Pa.
The present application further provides a kit comprising a composition provided herein. In some embodiments, the kit further comprises one or more sterile syringes. In some embodiments, the composition is preloaded into the one or more sterile syringes.
In some embodiments, the kit further comprises one or more sterile bandages. In some embodiments, the composition is preloaded onto a surface of the one or more sterile bandages.
In some embodiments, the kit further comprises one or more sterile surgical staples. In some embodiments, the composition is preloaded onto a surface of the one or more sterile surgical staples.
In some embodiments, the kit further comprises one or more sterile surgical sutures. In some embodiments, the composition is preloaded onto a surface of the one or more sterile surgical sutures.
The present application further comprises a sterile syringe comprising a composition provided herein.
The present application further comprises a sterile bandage, comprising a composition provided herein.
The present application further comprises a sterile surgical staple, comprising a composition provided herein.
The present application further comprises a sterile surgical suture, comprising a composition provided herein.
The present application further comprises a coating, comprising a pharmaceutically acceptable amount of a composition provided herein. In some embodiments, the coating is preloaded into a sterile syringe. In some embodiments, the coating is preloaded onto a surface of a sterile bandage. In some embodiments, the coating is preloaded onto a surface of a sterile surgical staple. In some embodiments, the coating is preloaded onto a surface of a sterile surgical suture.
The present application further provides a method of reducing or preventing tissue adhesion in a subject, comprising administering to the subject a composition provided herein.
In some embodiments, the tissue adhesion is associated with a surgical procedure, an injury, an anatomical defect, a cosmetic defect, or any combination thereof.
In some embodiments, the composition forms a barrier between two or more tissues in the subject, thereby reducing or preventing the tissue adhesion.
The present application further provides a method of reducing or preventing fibrotic adhesion formation in a subject, comprising administering to the subject a composition provided herein.
In some embodiments, the composition is administered by injection. In some embodiments, the composition is administered as a sprayable composition. In some embodiments, the composition is administered using a syringe.
In some embodiments, the method provided herein is associated with one or more of surgical, cosmetic, orthopedic, ophthalmic, and dermal applications.
In some embodiments, the composition provided herein is administered during a surgical procedure. In some embodiments, the surgical procedure is associated with one or more of abdominal, thoracic, pelvic, vascular, cardiovascular, neurological, and dermal surgical procedures.
In some embodiments, the method provided herein comprises local administration at a tissue associated with a surgical procedure, an injury, an anatomical defect, a cosmetic defect, or any combination thereof. In some embodiments, the surgical procedure is selected from the group consisting of a laparoscopic surgical procedure and an arthroscopic guided surgical procedure. In some embodiments, the surgical procedure is selected from the group consisting of laparoscopic myomectomy, ileal pouch anal-anastomosis, cesarean section, nerve repair, hernia repair, cardiac surgery, spinal surgery.
In some embodiments, the composition provided herein is administered using a catheter.
In some embodiments, the method comprises reducing or preventing the closure of permanent surgical central access lines, peripheral access lines, catheter lines, or drain lines, in the subject.
In some embodiments, the composition provided herein is locally administered at the site of a surgical suture or surgical staple in the subject.
In some embodiments, the composition forms a barrier between two or more tissues in the subject, thereby reducing or preventing the fibrotic adhesion formation.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.
A challenging medical problem associated with surgical intervention has been the formation of adhesions, as about 93% of patients who undergo open pelvic or abdominal surgery develop this pathology (see e.g., Menzies & Ellis, Ann. R. Coll. Surg. Engl. 1990, 72(1); 60-63; Parker et al, Colorectal. Dis. 2004, 6(6):506-511; and Parker et al, Dis. Colon Rectum 2001, 44(6):822-829). Postoperative adhesions are pathologic formations of fibrotic tissue that occur after peritoneal injury, and adhere the inner peritoneal lining of the abdominopelvic wall to internal organs within the abdominal or pelvic cavities (intestines, liver, gallbladder, urinary bladder, uterus, fallopian tubes, ovaries) (see e.g., Arung et al, World J. Gastroenterol. 2011, 17(41):4545-4553). A multifactorial cascade that involves ischemia, inflammation, angiogenesis, and tissue repair is known to cause its formation (see e.g., Reed et al, J. Surg. Res. 2002, 108(1):165-172; and Holmdahl et al, Eur. J. Surg. 1999, 165(11):1012-1019). Adhesions are associated with significant decreased quality of life, morbidity and mortality, and the incidence of hospital re-admissions due to its complications (intestinal obstruction, chronic abdominopelvic pain and secondary infertility) is as high as 20% (see e.g., Menzies & Ellis, Ann. R. Coll. Surg. Engl. 1990, 72(1); 60-63; Parker et al, Colorectal. Dis. 2004, 6(6):506-511; and Parker et al, Dis. Colon Rectum 2001, 44(6):822-829; Ouaissi et al J. Visc. Surg. 2012, 149(2), e104-114; Klingensmith et al, Surg. Endosc. 1996, 10(11):1085-7; Hallfeldt et al, Zentralbl. Chir. 1995, 120(5):387-391; Kolmorgen et al, Zentralbl. Gyakol. 1991, 113(6): 291-295; Swank et al, Lancet 2003, 361(9365):1247-1251; Tulandi et al, Am. J. Obstet. Gynecol. 1990, 162(2):354-357; Marana et al, Hum. Reprod. 1999, 14(12):2991-2995; Milingos et al, Ann. N.Y. Acad. Sci. 2000, 900:272-285; and Vrijland et al, Surg. Endosc. 2003, 17(7):1017-1022.
Physical barriers in the form of films have been reported for use in preventing adhesions, however, their application to irregular surfaces and cavities is challenging or impossible, as the films are fragile, difficult to handle, incompatible with minimally invasive laparoscopic or catheter-based procedures, and limited efficacy (e.g., ˜25%) decreases clinical adoption (see e.g., Wilson, M. S. Colorectal. Dis. 2007, 9 Suppl. 2:60-65; and Hirschelmann et al, Arch. Gynecol. Obster. 2012, 285(4):1089-1097). The development of new technologies to solve these clinical limitations is desirable.
Hydrogel formulations for the prevention of postoperative adhesions may be particularly attractive as a substitute for commercially available barriers. Without being bound by theory, an exemplary hydrogen biomaterial would include mechanical and biological properties to prevent cell adherence, infiltration, and adhesion formation. The hydrogel would also be injectable to be compatible with minimally invasive procedures such as laparoscopies and arthroscopies, and sprayable to uniformly cover large and irregular areas during laparotomies or thoracotomies, a limitation that current ‘film-based’ barriers have.
To achieve the desired mechanical and biological properties, the present application provides a hydrogel composed of silicate nanoplatelets (SNP) and poly(ethylene oxide) (PEO). The disc-shaped SNPs (e.g., thickness ˜0.92 nm and diameter ˜25 nm) possess unique electrostatic properties with negatively charged surfaces and positively charged edges that result in a nanoscale surface-to-edge attraction and spontaneous formation of a superstructure. The dual electrostatic charges in the surface of SNPs nanoplatelets confer non-Newtonian and shear-thinning behavior to the material, allowing its injectability and sprayability after subjecting the material to stress, with subsequent mechanical recovery immediately after delivery (
Without being bound by theory, it was hypothesized that the synergistic combination of the mechanical and biological features of SNPs and PEO would provide a shear-thinning hydrogel barrier (STHB) that can be administered via multiple facile delivery methods (e.g., spraying, injecting, spreading, and the like). The compositions provided herein were designed to provide physical separation between tissues and inhibit infiltration of collagen-secreting cells that lead to adhesion formation, thereby providing a universal solution to prevent the formation of postoperative adhesions over a wide range of surgical procedures.
Accordingly, the present application provides compositions comprising silicate nanoplatelets (SNPs) and poly(ethylene oxide). In some embodiments, the present application provides compositions comprising laponite (an exemplary SNP) and poly(ethylene oxide). In some embodiments, the compositions provided herein exhibit shear-thinning behavior (i.e., the compositions are shear-thinning compositions). The expression “shear-thinning” or “shear-thinning behavior”, refers to a decrease in viscosity (i.e., increasing flow rate) of a composition with increasing application of shear stress. For example, a shear-thinning composition (i.e. a composition exhibiting shear-thinning behavior) can exhibit a decrease in viscosity (i.e. increase in flow) upon application of an increasing rate of shear stress.
In some embodiments, the composition comprises about 3% to about 12% by weight laponite, for example, about 3% to about 10%, about 3% to about 8%, about 3% to about 5%, about 5% to about 12%, about 5% to about 10%, about 5% to about 8%, about 8% to about 12%, about 8% to about 10%, or about 10% to about 12% by weight laponite. In some embodiments, the composition comprises about 3% to about 10% laponite. In some embodiments, the composition comprises about 5% to about 10% laponite. In some embodiments, the composition comprises about 8% to about 10% laponite.
In some embodiments, the laponite comprises a positively charged edge and a negatively charged surface. In some embodiments, the overall charge of the laponite is negative.
In some embodiments, the laponite has an average diameter of from about 5 nm to about 60 nm, for example, about 5 to about 40, about 5 to about 20, about 5 to about 10, about 10 to about 60, about 10 to about 40, about 10 to about 20, about 20 to about 60, about 20 to about 40, or about 40 to about 60 nm. In some embodiments, the laponite has an average diameter of from about 10 nm to about 40 nm. In some embodiments, the laponite has an average diameter of from about 20 nm to about 30 nm.
In some embodiments, the laponite has an average thickness of from about 0.5 nm to about 2 nm, for example, about 0.5 to about 1.5, about 0.5 to about 1, about 0.5 to about 0.75, about 0.75 to about 2, about 0.75 to about 1.5, about 0.75 to about 1, about 1 to about 2, about 1 to about 1.5, or about 1.5 to about 2 nm. In some embodiments, the laponite has an average thickness of about 1 nm.
In some embodiments, the laponite provided herein comprises about 66% SiO2, about 30% MgO, about 3% Na2O, and about 1% LiO2. In some embodiments, the laponite provided herein consists of about 66% SiO2, about 30% MgO, about 3% Na2O, and about 1% LiO2.
In some embodiments, the composition comprises about 0.5% to about 10% by weight poly(ethylene oxide) (PEO), for example, about 0.5% to about 8%, about 0.5% to about 6%, about 0.5% to about 5%, about 0.5% to about 4%, about 0.5% to about 3%, about 0.5% to about 2%, about 0.5% to about 1%, about 1% to about 10%, about 1% to about 8%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, about 2% to about 10%, about 2% to about 8%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, about 3% to about 10%, about 3% to about 8%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 10%, about 4% to about 8%, about 4% to about 6%, about 4% to about 5%, about 6% to about 10%, about 6% to about 8%, or about 8% to about 10%, by weight poly(ethylene oxide). In some embodiments, the composition comprises about 1% to about 5% by weight poly(ethylene oxide). In some embodiments, the composition comprises about 1% to about 3% by weight poly(ethylene oxide). In some embodiments, the composition comprises about 3% to about 5% by weight poly(ethylene oxide).
In some embodiments, the poly(ethylene oxide) has an average molecular weight of about 18,000 to about 22,000 g/mol, for example, about 18,000 to about 21,000, about 18,000 to about 20,000, about 18,000 to about 19,000, about 19,000 to about 22,000, about 19,000 to about 21,000, about 19,000 to about 20,000, about 20,000 to about 22,000, about 20,000 to about 21,000, about 21,000 to about 22,000 g/mol.
In some embodiments, the composition comprises:
about 3% to about 10% by weight laponite; and
about 1% to about 10% by weight poly(ethylene oxide).
In some embodiments, the composition comprises:
about 5% to about 10% by weight laponite; and
about 1% to about 5% by weight poly(ethylene oxide).
In some embodiments, the composition comprises:
about 8% to about 10% by weight laponite and about 1% to about 5% by weight poly(ethylene oxide); or
about 5% to about 10% by weight laponite and about 1% to about 3% by weight poly(ethylene oxide); or
about 8% to about 10% by weight laponite and about 1% to about 3% by weight poly(ethylene oxide).
In some embodiments, the composition comprises:
about 5% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 5% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 5% by weight laponite and about 3% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 3% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 3% by weight poly(ethylene oxide).
In some embodiments, the composition comprises:
about 5% by weight laponite and about 1% by weight poly(ethylene oxide); or
about 8% by weight laponite and about 2% by weight poly(ethylene oxide); or
about 10% by weight laponite and about 3% by weight poly(ethylene oxide).
In some embodiments, the composition comprises about 10% by weight laponite and about 3% by weight poly(ethylene oxide).
In some embodiments, the compositions provided herein further comprise water.
In some embodiments, the composition comprises about 80% to about 95% by weight water, for example, about 80% to about 90%, about 80% to about 85%, about 85% to about 95%, about 85% to about 90%, or about 90% to about 95% by weight water. In some embodiments, the composition comprises about 85% to about 94% by weight water. In some embodiments, the water is deionized water. In some embodiments, the water is ultra-pure water (e.g., Milli-Q) or buffered water (e.g., phosphate buffered saline).
In some embodiments, the composition comprises:
about 8% to about 10% by weight laponite, about 1% to about 5% by weight poly(ethylene oxide), and about 85% to about 91% by weight water; or
about 5% to about 10% by weight laponite, about 1% to about 3% by weight poly(ethylene oxide), and about 87% to about 94% by weight water; or
about 8% to about 10% by weight laponite, about 1% to about 3% by weight poly(ethylene oxide), and about 87% to about 91% by weight water.
In some embodiments, the composition provided herein consists of laponite, poly(ethylene oxide), and water.
In some embodiments, the composition provided herein is a gel. In some embodiments, the composition provided herein is a hydrogel. As used herein, the term “hydrogel” refers to a gel in which the liquid component comprises water.
The present application further provides a process of preparing a composition provided herein, the process comprising:
(a) combining laponite and water to form a first mixture; and
(b) adding the poly(ethylene oxide) to the first mixture to form the composition.
In some embodiments, the composition provided herein is prepared according to a process described herein. For example, in some embodiments, the present application provides a compositions comprising about 5% to about 10% by weight laponite, about 1% to about 5% by weight poly(ethylene oxide), and about 85% to about 94% by weight water, wherein the composition is prepared according to a process comprising:
(a) combining the laponite and water to form a first mixture;
(b) adding the poly(ethylene oxide) to the first mixture to form the composition.
In some embodiments, the yield stress of the composition provided herein is from about 100 Pa to about 2000 Pa, for example, about 100 Pa to about 1500 Pa, about 100 Pa to about 1000 Pa, about 100 Pa to about 750 Pa, about 100 Pa to about 500 Pa, about 100 Pa to about 250 Pa, about 100 Pa to about 200 Pa, about 200 Pa to about 2000 Pa, about 200 Pa to about 1500 Pa, about 200 Pa to about 1000 Pa, about 200 Pa to about 750 Pa, about 200 Pa to about 500 Pa, about 200 Pa to about 250 Pa, about 250 Pa to about 2000 Pa, about 250 Pa to about 1500 Pa, about 250 Pa to about 1000 Pa, about 250 Pa to about 750 Pa, about 250 Pa to about 500 Pa, about 500 Pa to about 2000 Pa, about 500 Pa to about 1500 Pa, about 500 Pa to about 1000 Pa, about 500 Pa to about 750 Pa, about 750 Pa to about 2000 Pa, about 750 Pa to about 1500 Pa, about 750 Pa to about 1000 Pa, about 1000 Pa to about 2000 Pa, about 1000 Pa to about 1500 Pa, or about 1500 Pa to about 2000 Pa.
In some embodiments, a shear-thinning composition provided herein flows upon application of a pressure greater than the yield stress, for example, application of a pressure about 10% greater, about 20% greater, about 30% greater, about 40% greater, about 50% greater about 60% greater, about 70% greater, about 80% greater, about 90% greater, or about 100% greater than the yield stress.
In some embodiments, a composition provided herein is self-healing. As used herein, the expression “self-healing”, refers to recovery of the elastic gel strength of a composition upon removal of a stress. In some aspects, a self-healing composition may recover elastic gel strength from about 2 seconds to about 1 minute after removal of a stress, for example, from 30 seconds to 1 min., from 30 seconds to 45 seconds, from 15 seconds to 1 minute, from 15 seconds to 45 seconds, from 15 seconds to 30 seconds, from 10 seconds to 15 seconds, from 10 seconds to 30 seconds, from 10 seconds to 45 seconds, from 10 seconds to 1 minute, from 5 seconds to 10 seconds, from 5 seconds to 25 seconds, from 5 seconds to 45 seconds, from 5 seconds to 1 minute, from 2 seconds to 10 seconds, from 2 seconds to 25 seconds, from 2 seconds to 45 seconds, or from about 2 seconds to 1 minute.
As used herein, the term “consisting essentially of” is used to define a composition or method that includes materials, steps, features, components, or elements, in addition to those explicitly provided herein, provided that these additional included materials, steps, features, components, or elements do not materially affect the basic and novel characteristic(s) of the claimed invention. In some embodiments, the additional materials, steps, feature, components, or elements do not affect the mode of action to achieve the desired result of the invention described herein. The term “consists essentially of” or “consisting essentially of” occupies a middle ground between “comprising” and “consisting of”.
In some embodiments, the present application provides a composition consisting essentially of silicate nanoplatelets (SNPs), poly(ethylene oxide), and one or more additional components. In some embodiments, the present application provides a composition consisting essentially of laponite, poly(ethylene oxide), and one or more additional components. In some embodiments, the present application provides a composition consisting essentially of silicate nanoplatelets (SNPs), poly(ethylene oxide), water, and one or more additional components (e.g., one or more excipients, buffering agents, additional therapeutic agents, and the like). In some embodiments, the one or more additional components do not materially affect the shear-thinning characteristics of the compositions described herein. In some embodiments, the one or more additional components do not materially affect the anti-adhesion characteristics of the compositions described herein. In some embodiments, the one or more additional components do not materially affect the shear-thinning and anti-adhesion characteristics of the compositions described herein.
For the terms “for example” and “such as” and grammatical equivalences thereof, the phrase “and without limitation” is understood to follow unless explicitly stated otherwise. As used herein, the term “about” is meant to account for variations due to experimental error. All measurements reported herein are understood to be modified by the term “about”, whether or not the term is explicitly used, unless explicitly stated otherwise. As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.
The following abbreviations may be used throughout the present application: STHB: Shear-Thinning Hydrogel Barrier; SNP: Silicate Nanoplatelets (e.g., laponite); PEO: Poly(ethylene oxide); FDA: Food and Drug Administration; LVR: Linear Viscoelastic Region; PTFE: Polytetrafluoroethylene; PA: Peritoneal Adhesion Index; H&E: Hematoxylin and eosin.
The present application further provides methods of reducing (e.g., reducing the likelihood) and/or preventing tissue adhesion in a subject. In some embodiments, the method comprises administering the composition to the subject (e.g., an amount effective to reduce and/or prevent tissue adhesion in the subject). In some embodiments, the method is a method of reducing tissue adhesion in the subject. In some embodiments, the method is a method is preventing tissue adhesion in the subject.
As used herein, the term “subject,” refers to any animal, including mammals. For example, mice, rats, other rodents, guinea pigs, rabbits, dogs, cats, swine, cattle, sheep, horses, primates, and humans. In some embodiments, the subject is a human.
In some embodiments, the tissue adhesion is associated with a surgical procedure, an injury, an anatomical defect, a cosmetic defect, or any combination thereof. In some embodiments, the tissue adhesion is associated with a surgical procedure. In some embodiments, the tissue adhesion is associated with an injury. In some embodiments, the tissue adhesion is associated with an anatomical defect. In some embodiments, the tissue adhesion is associated with a cosmetic defect.
In some embodiments, the composition provided herein, upon administration to the subject, forms a barrier between two or more tissues in the subject, thereby reducing or preventing the tissue adhesion.
The present application further provides a method of reducing or preventing fibrotic adhesion formation in a subject. In some embodiments, the method comprises administering the composition to the subject (e.g., an amount effective to reduce and/or prevent fibrotic adhesion formation in the subject). In some embodiments, the method is a method of reducing fibrotic adhesion formation in the subject. In some embodiments, the method is a method is preventing fibrotic adhesion formation in the subject. In some embodiments, the composition forms a barrier between two or more tissues in the subject, thereby reducing or preventing the fibrotic adhesion formation.
In some embodiments, the method provided herein is associated with one or more of surgical, cosmetic (e.g., a cosmetic surgical procedure), orthopedic, ophthalmic, and dermal applications. In some embodiments, the composition is administered during a surgical procedure.
In some embodiments, the surgical procedure is associated with one or more procedures selected from abdominal, thoracic, pelvic, vascular, cardiovascular, neurological, and dermal surgical procedures. In some embodiments, the surgical procedure is selected from the group consisting of a laparoscopic surgical procedure and an arthroscopic guided surgical procedure. In some embodiments, the surgical procedure is selected from the group consisting of laparoscopic myomectomy, ileal pouch anal-anastomosis, cesarean section, nerve repair, hernia repair, cardiac surgery, spinal surgery.
In some embodiments, the method comprises local administration at a tissue associated with a surgical procedure, an injury, an anatomical defect, a cosmetic defect, or any combination thereof. In some embodiments, the method comprises local administration at a tissue associated with a surgical procedure. In some embodiments, the method comprises local administration at a tissue associated with an injury. In some embodiments, the method comprises local administration at a tissue associated with an anatomical defect. In some embodiments, the method comprises local administration at a tissue associated with a cosmetic defect.
As used here, the term “local administration” refers to administration at or within close proximity to the site at which the tissue adhesion is to be reduced and/or prevented in the subject. For example, upon cessation of the administration, a composition provided herein will remain substantially localized at the site of the administration. In some embodiments, the composition is locally administered at the site of a surgical suture or surgical staple in the subject.
In some embodiments, the composition is administered onto one or more surfaces of the subject (e.g., one or more tissues, one or more organs, and the like). In some embodiments, the composition is administered onto one or more internal surfaces of the subject (e.g., one or more tissues, one or more organs, and the like). In some embodiments, the composition is administered onto the skin of the subject.
In some embodiments, the method comprises reducing or preventing the closure of permanent surgical central access lines, peripheral access lines, catheter lines, or drain lines, or any combination thereof, in the subject.
The amount of the composition administered to a subject will vary depending upon the composition being administered, the purpose of the administration, the state of the subject, the manner of administration, and the like. Effective doses will depend on the necessary treatment as well as by the judgment of the attending clinician depending upon factors such as the type and severity of the complication, the age, weight and general condition of the subject, and the like.
The compositions provided herein may also be useful as components in cosmetic compositions. Benefits of shear-thinning compositions in cosmetic applications include, but are not limited to, the biocompatibility and non-toxic character of the compositions provided herein (e.g., a shear-thinning composition may degrade over time in subcutaneous tissue). Exemplary cosmetic applications include, but are not limited to, cosmetic surgical procedures such as dental augmentation, breast augmentation, buttock augmentation, lip augmentation, jaw augmentation, hip augmentation, chin augmentation, brow augmentation, arm augmentation, leg augmentation, and the like. In some embodiments, the composition provided herein is administered during a cosmetic surgical procedure.
The compositions provided herein can be administered by a variety of routes, depending upon the area to be treated. In some embodiments, the composition is an injectable composition and/or a sprayable composition. In some embodiments, the composition is an injectable composition. In some embodiments, the composition is a sprayable composition.
In some embodiments, the composition is administered by parenteral administration. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal intramuscular, injection, or infusion; or intracranial, (e.g., intrathecal or intraventricular administration).
In some embodiments, the composition is administered by injection. In some embodiments, the composition is administered using a syringe. In some embodiments, the composition is administered using a catheter.
In some embodiments, the composition is preloaded onto at least one surface of a medical device (e.g., a syringe, catheter, surgical stable, medical wipe, bandage, a device suitable for spraying the composition (e.g., a container substantially similar to the container shown in
The present application further provides a medical device comprising a compound provided herein. Exemplary medical derives include, but are not limited to syringes, staples, bandages, catheters, and the like. The present application further provides a syringe (e.g., a sterile syringe) comprising a composition provided herein. The present application further provides a bandage (e.g., a sterile bandage) comprising a composition provided herein. The present application further provides a staple (e.g., a sterile staple or a sterile surgical staple) comprising a composition provided herein. In some embodiments, the staple is first applied to the subject, and the composition is subsequently administered to one or more surfaces of the staple. The present application further provides a suture (e.g., a sterile suture or a sterile surgical suture) comprising a composition provided herein. In some embodiments, the suture is first applied to the subject, and the composition is subsequently administered to one or more surfaces of the suture.
In some embodiments, the compositions provided herein are administered in the form of a coating (e.g., a spreadable coating). In some embodiments, the coating is preloaded into a syringe (e.g., a sterile syringe). In some embodiments, the coating is preloaded onto a surface of a bandage (e.g., a sterile bandage). In some embodiments, the coating is preloaded onto a surface of a staple (e.g., a sterile staple or a sterile surgical staple). In some embodiments, the coating is preloaded onto a surface of a suture (e.g., a sterile suture or a sterile surgical suture).
The present application further provides a kit comprising a composition provided herein. In some embodiments, the kit further comprises one or more medical devices described herein. In some embodiments, the components of the kit can be separately packaged or contained.
In some embodiments, the kit further comprises one or more syringes (e.g., one or more sterile syringes). In some embodiments, the composition is preloaded into the one or more syringes. In some embodiments, the composition is preloaded into the one or more devices suitable for spraying the composition.
In some embodiments, the kit further comprises one or more bandages (e.g., one or more sterile bandages). In some embodiments, the composition is preloaded onto a surface of the one or more bandages.
In some embodiments, the kit further comprises one or more staples (e.g., one or more sterile staples). In some embodiments, the composition is preloaded onto a surface of the one or more staples.
In some embodiments, the kit further comprises one or more sutures (e.g., one or more sterile sutures or one or more sterile surgical sutures). In some embodiments, the composition is preloaded onto a surface of the one or more sutures.
In some embodiments, the kit further comprises one or more catheters (e.g., one or more sterile catheters). In some embodiments, the composition is preloaded into the one or more catheters.
Instructions, either as inserts or as labels, indicating quantities of the composition to be administered, guidelines for administration, guidelines for mixing components of the composition, and/or guidelines for preparing a composition according to a process described herein, can also be included in a kit provided herein. In some embodiments, the instructions further comprise instructions for performing one or more of the methods provided herein. In some embodiments, the instructions further comprise instructions for quantities of the composition to be administered, guidelines for administration, guidelines for mixing components of the composition, and/or guidelines for preparing a composition according to a process described herein.
The kits provided herein can further include, if desired, one or more conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art.
The invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes, and are not intended to limit the invention in any manner.
3T3 fibroblasts (ATCC, Manassas, Va.) were cultured in Dulbecco's modified eagle medium (DMEM) (Invitrogen, Carlsbad, Calif.) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Flowery Branch, Ga.) and 1% penicillin-streptomycin (PS) (MediaTech Inc., Manassas, Va.) under 5% CO2 at 37° C. Cells were passaged approximately 2 times per week and media was exchanged every 2 days.
All the results are expressed as mean±SD. An unpaired Student t test was used to determine statistical significance of all samples and groups. *P<0.05, **P<0.01 will be considered statistically significant.
Biomaterials with shear-thinning properties are essential for the development of viscoelastic gel coatings that transform their mechanical properties to liquid-like states upon delivery under shear stress. This property enables their injectability or sprayability as a flowable liquid, and subsequent recovery to their original viscoelastic solid state when shear is removed after delivery. Previous attempts to provide sprayable hydrogel coatings as adhesion barriers have employed in-situ chemistry (polymerization or crosslinking) to achieve the desired viscoelastic properties of hydrogel coatings delivered from a liquid precursor formulation. The STHB properties described herein enable the development of injectable and sprayable hydrogel compositions for standard and minimally invasive medical interventions without the need of additional polymerization or crosslinking (see e.g, Gaharwar et al, ACS Nano. 2014, 8(10):9833-9842; and Avery et al, Sci. Transl. Med. 2016, 8(365):365ra156). STHB technology achieves these goals with far greater simplicity and elegance than 2-part in-situ crosslinked chemical systems.
STHBs were formulated with different percentages of poly(ethylene oxide) (PEO) (Sigma-Aldrich, St. Louis, Mo.) and silicate nanoplatelets (SNPs) Laponite XLG (BYK, Wesel, Germany). PEO and SNPs were sterilized using UV light and dissolved in ultra-filtered deionized water in separate vials. For dissolution, SNPs were stirred (400 rpm) at 60° C., and after 1 minute of stirring, PEO solution was added and stirred for four additional minutes until complete homogenization was achieved. The hydrogel compositions that were fabricated are 5 wt % SNP (5L), 8 wt % SNP (8L), 10 wt % SNP (10L), 5 wt % SNP 1 wt % PEO (5L1P), 8 wt % SNP 1 wt % PEO (8L1P), 10 wt % SNP 1 wt % PEO (10L1P), 5 wt % SNP 2 wt/o PEO (5L2P), 8 wt % SNP 2 wt % PEO (8L2P), 10 wt % SNP 2 wt % PEO (10L2P), 5 wt % SNP 3 wt % PEO (5L3P), 8 wt % SNP 3 wt % PEO (8L3P), and 10 wt % SNP 3 wt % PEO (10L3P) (see e.g., (
After formulation, the rheological properties of all STHB compositions were determined, and higher viscosity and stronger gel formation was observed on formulations with higher SNP concentrations (see e.g.,
To characterize the rheology and mechanical properties of STHB formulations, a MCR 301 rheometer was used (Anton Paar, Graz, Austria) as previously described (see e.g., Gaharwar et al, ACS Nano. 2014, 8(10):9833-9842). Temperature sweeps were performed on a 25-mm diameter plate (gap height: 500 μm), and mineral oil was placed around to prevent water evaporation. Equilibration time was set to 10 min before testing, followed by steady shear at 10 s−1 for 2 minutes. Shear rate sweeps (0.001 to 100 s−1 with 10 points/decade) and strain sweeps (0.01 to 1000% at 1 Hz) were performed at 37° C. Recovery testing was performed by applying a value outside of the linear viscoelastic range (100% strain), followed by a value inside of the linear viscoelastic range (1% strain) at 1 Hz.
It was found that 10L3P presented higher viscosity (˜727 Pa·s) compared to formulations with lower SNP concentration, where 5L3P and 8L3P displayed a viscosity of ˜81 Pa·s and ˜436 Pa·s when subjected to a shear of ˜1.6 s−1, as shown in
The injection force required to extrude STHB formulations was analyzed using a mechanical tester (Instron Model 5542) (Instron, Norwood, Mass.). STHB formulations were loaded into 3-mL syringes (BD Biosciences, San Jose, Calif.) and injected through 3 different intraluminal diameter needle sizes, 0.838 mm (18 G), 0.337 mm (23 G), and 0.210 mm (27 G) (BD Biosciences, San Jose, Calif.). The syringe plungers were pressed by an upper compressive platen, and the lower housing of the syringe was placed into the tensile grip of the instrument to prevent movement. The injection rate used was 2 mL/min and the force on the plunger was measured with a 100-N load cell. All STHB samples were tested in triplicate. Bluehill version 3 software (Instron, Norwood, Mass.) was used to analyze the data.
The viscoelastic behavior of STHBs was further characterized by strain sweeps (0.01 to 1000% at 1 Hz). The linear viscoelastic region (LVR) of STHBs was obtained within a small strain region (0.1-10.0 strain (%) at 1 Hz), as shown in
Low (1%) and high (100%) strain at 1 Hz was applied to STHBs over multiple cycles, demonstrating self-recovery to their original modulus, as shown in
To determine the sprayability of STHB formulations and facilitate visualization, hydrogel compositions were labeled with Alexa Fluor 488 dye (Thermo Fisher Scientific, Waltham, Mass.). STHB formulations were loaded into 10 mL syringes (BD, Franklin Lakes, N.J.) and sprayed through a specialized syringe extrusion setup was mounted on a mechanical testing instrument as shown in
The injectability of STHBs was evaluated on three needles with different intraluminal diameters (18 G, 23 G, 27 G). Test parameters are presented in Table 2.
The force needed to extrude the hydrogel was linearly increased until it reached a plateau defining the maximum extrusion force in each formulation, as shown in
A spraying system equipped with a nozzle size of 22 G was used to spray the 5L3P, 8L3P, and 10L3P formulations, and average sprayed area and spot size was quantified as previously reported, as shown in
To further study STHBs topical applications, a spreading analysis was performed as previously reported (see e.g., Lardy et al, Drug Development and Industrial Pharmacy 2000, 26(7):715-721). The spreadability of the hydrogels was evaluated 48 hours after their preparation. STHB formulations were incubated for 1 hour at 37° C., and after incubation, the studies were quickly performed at room temperature. STHBs were placed between two horizontal transparent glass plates (Bio-Rad, Hercules, Calif.), and a 125 gram weight was placed on the upper plate. After one minute, the weight was removed, and the spreading diameter measured.
Spreadability was quantified as the total diameter covered by the hydrogel within the plates, and its fluidity and stiffness classified according to a previous published work (see e.g., Lardy et al, Drug Development and Industrial Pharmacy 2000, 26(7):715-721). The spreadability was quantified by placing hydrogels in a device where a standard weight (125 grams) was applied, as shown in
The resulting spread diameter of STHB compositions with 5 wt % SNP was in the range of 37.9 to 45.9 mm, 26.9 to 31.4 mm for 8 wt % SNP compositions, and 23.3 to 27.5 mm for 10 wt % SNP compositions, respectively, as shown in
To determine the stability and swelling ratio of STHB formulations, one gram of each formulation was placed in a cell strainer (Corning, Corning, N.Y.) (n=3). Each strainer was submerged in 7 mL of PBS on 6-well plates (Corning, Corning, N.Y.) and incubated at 37° C. Stability and swelling were recorded at 3, 7, 14, and 21 days by quantifying the wet weight and dry weight after lyophilization of STHB compositions. Stability was calculated with the formula: mass loss percentage=(M0−Md)/M0×100%, and swelling ratio (Q) with the formula: Q=(Ms−Md)/Md (see e.g., Tang et al, Journal of Applied Polymer Science 2007, 104(5):2785-2791). M0 represents the original mass of the hydrogel before immersing it into the medium, Ms is the mass of the hydrogel in the swollen state and Md is the mass of the hydrogel in the dry state.
After 21 days, the mass loss of STHB formulations was below 3%, as shown in
Prevention of cell adherence and infiltration to the hydrogel formulations is fundamental for the creation of an effective adhesion barrier. For this purpose, the cell-material interactions between STHBs and the effector cells (fibroblasts) of the formation of postoperative adhesions were investigated.
To determine cellular adherence to the hydrogel compositions, 3T3 fibroblasts were seeded on hydrogel films. 24-well culture plates (Corning Inc., Corning, N.Y.) were coated with 0.2 mL of each hydrogel formulation, and 10×104 cells were seeded. 24 hours after seeding, fibroblasts were washed with PBS to remove unattached cells. Remaining fibroblasts were detached with trypsin (Sigma-Aldrich, Darmstadt, Germany) for posterior quantification. Fibroblast numbers were determined by PrestoBlue Cell Viability Reagent (Thermo Fisher Scientific, Waltham, Mass.) via a microplate reader (BioTek Synergy 2, Winooski, Vt.). Analysis was performed by using BioTek Gen5 software (BioTek Synergy 2, Winooski, Vt.).
As a control, cells were seeded on polytetrafluoroethylene (PTFE) coated substrates suitable for cell adherence and compared to cells seeded on STHB surfaces, as shown in
Cellular morphology was evaluated after seeding 10×104 3T3 fibroblast on the surface of STHB formulations placed on 6-well culture plates (Corning Inc., Corning, N.Y.). After being incubated for 24 hours, cells were fixed using a 4% paraformaldehyde solution (Sigma-Aldrich, St. Louis, Mo.), followed by F-actin (red) (Thermo Fisher Scientific, Waltham, Mass.) and DAPI (Sigma-Aldrich, St. Louis, Mo.) staining. Cellular fluorescent micrographs at different locations of the material surfaces were obtained using a fluorescence microscope (Zeiss, Oberkochen, Germany), and analyzed by Snap 2058-Zen Pro 2012 software (Zeiss, Oberkochen, Germany). Sixty individual cells per group were randomly selected in each micrograph for analysis. The maximum orthogonal length, width and area of each cell was measured using ImageJ software (National Institutes of Health, Bethesda, Md.), and the aspect ratio was calculated as the longer length divided by the shorter length.
The aspect ratio (based on dimensions, geometry and area) of single cells was quantified to understand cell adherence and expansion (see e.g., Collins et al, Proc. Natl. Acad. Sci. U.S.A. 2017, 114(29):E5835-E5844). Single-cell analysis (aspect ratio and morphology) was performed via F-actin fluorescence labeling. Fluorescent micrographs showed that cells seeded on PTFE substrates (control) and SNP-only formulations (5L, 8L, 10L), had normal cell adherence and pseudopodia expansion, as shown in
Cells in control and SNP only groups presented a disparate set of aspect ratios on the graphs shown in
Cytotoxicity of STHB formulations was evaluated, 10×103 3T3 fibroblasts were seeded in 96-well culture plates (Corning Inc., Corning, N.Y.) and incubated for 48 hours with the following ranges of SNPs, PEO and SNPs combined with PEO: 0.001 to 1000 μg/mL. After the incubation period, cell viability was quantified by PrestoBlue Cell Viability Reagent (Thermo Fisher Scientific, Waltham, Mass.) via a microplate reader (BioTek Synergy 2, Winooski, Vt.) and BioTek Gen5 software (BioTek Synergy 2, Winooski, Vt.).
Biocompatibility of STHB compositions was assessed on fibroblasts treated with SNPs, PEO, and a combination of both components (0.001 to 1000 μg/mL). As shown in
To investigate STHB efficacy in preventing postoperative adhesions, a peritoneal injury rat model (300 g male Wistar rats (Charles River Laboratories, Worcester, Mass.) with 8 ischemic peritoneal buttons was used (
The Peritoneal Adhesion Index (PAI) was used as a scoring system to grade the adhesions based on several morphological features such as vascularization, thickness, strength, and damage, as shown in
The severity of adhesions was graded with the following scoring system: 0—no adhesion, 1—filmy adhesion that needs blunt dissection, 2—strong adhesion that needs sharp dissection, 3—very strong vascularized adhesion that needs sharp dissection with damage hardly preventable. Each button (8) was individually graded and the index was calculated based on the sum of the total score of the eight buttons. Percentage of adhesion formation and efficacy of adhesion prevention was calculated based on the number of adhesions formed, each button was considered as 12.5% of the total number of injuries created (8 ischemic buttons).
In the control group, adhesions were found attached to the ischemic button and its surroundings. Seprafilm®, 5L3P, and 8L3P groups had lower adhesion grades, whereas 10L3P group did not develop any observable adhesions, as shown in
To perform a quantitative analysis of the severity of adhesion formation in all the groups, parameters such as adhesion grade, number, percentage of adhesion formation and efficacy of adhesion prevention were examined. The total average PAI score per group was 9.6±0.5 for control, 4.6±0.4 for Seprafilm®, 6.6±1.9 for 5L3P, 2.4±1.4 for 8L3P, and 0 for 10L3P, as shown in
The average percentage of adhesion formation per group was 70%±3 for control, 50%±5 for Seprafilm®, 38%+8 for 5L3P, 15% 7 for 8L3P, and 0% for 10L3P, as shown in
After 14 days, histopathological examination was performed to evaluate tissue remodeling, inflammatory response and STHB absorption. Peritoneal button tissues were extracted, frozen, and sectioned into 7 μm sagittal and transversal slices using a HM550 Cryostat system (Thermo Fischer Scientific, Waltham, Mass.). Sections were stained with Hematoxylin & Eosin (Sigma-Aldrich, St. Louis, Mo.) and Masson's trichrome (Sigma-Aldrich, St. Louis, Mo.) to assess tissue morphological changes and fibrotic formation. Anti-CD68 (Abcam, Cambridge, Mass.), and anti-CD3 (Abcam, Cambridge, Mass.) primary antibodies with Alexa Fluor-conjugated (Invitrogen, Carlsbad, Calif.) secondary antibodies, were used in conjunction with DAPI (Vector Laboratories, Burlingame, Calif.) to perform immunohistology. Slides were examined (n=5 pictures per section) using an Axio Observer microscope (Zeiss, Oberkochen, Germany).
As shown in
The abdominal cavity of the animal was also examined and no hydrogel or Seprafilm® barrier was found. H&E examination confirmed its degradation in a period of 2 weeks. Rapid resorption is an important requirement for an adhesion barrier material, as lengthy residence time in the peritoneal cavity can retard re-mesothelialization of the peritoneal lining.
Masson's trichrome staining was also performed to determine the presence of fibrotic bands in the peritoneal wall, as shown in
Local inflammatory response was analyzed by CD3 and CD68 immunostaining, as shown in
These observations confirmed that STHB formulations were superior in preventing adhesions compared to a commercially available barrier, and biocompatible as no abnormal local immunological response was observed.
It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
This application claims the benefit of U.S. Provisional Application Ser. No. 62/677,350, filed May 29, 2018, the disclosure of which is incorporated herein by reference in its entirety.
This invention was made with Government support under Grant Nos. 1R01EB023052, 1R01HL140618, 1R01HL137193, and 1R01GM126831, awarded by the National Institutes of Health. The Government has certain rights in the invention.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2019/034298 | 5/29/2019 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 62677350 | May 2018 | US |
