Multi-layer biomaterial for tissue regeneration and wound healing

Abstract

The technology described herein is directed to compositions comprising at least a first porous biomaterial layer and a second impermeable biomaterial layer and methods relating thereto. In some embodiments, the compositions and methods described herein relate to wound healing, e.g. repair of wounds and/or tissue defects.

What is claimed herein is: 
     
         1 . A composition comprising a first and second layer;
 the first layer comprising a porous biomaterial matrix; and   the second layer comprising a impermeable biomaterial.   
     
     
         2 . The composition of  claim 1 , wherein the matrix is a structure selected from the group consisting of:
 foams; hydrogels; electro spun fibers; gels; fiber mats; sponges; 3-dimensional scaffolds; non-woven mats; woven materials; knit materials; fiber bundles;   and fibers.   
     
     
         3 . The composition of any of  claims 1 - 2 , wherein the biomaterial is selected from the group consisting of:
 silk fibroin; PGA; collagen; polyethylene oxide, collagen, fibronectin, keratin, polyaspartic acid, polylysin, alginate, chitosan, chitin, and hyaluronic acid.   
     
     
         4 . The composition of any of  claims 1 - 3 , wherein the silk fibroin comprises  Bombyx mori  silk fibroin. 
     
     
         5 . The composition of any of  claims 1 - 4 , wherein the first and second layers comprise the same biomaterial. 
     
     
         6 . The composition of any of  claims 1 - 4 , wherein the first and second layers comprise different biomaterials. 
     
     
         7 . A composition comprising a first and second layer;
 the first layer comprising a porous silk fibroin matrix; and   the second layer comprising a impermeable silk fibroin film.   
     
     
         8 . The composition of any of  claims 1 - 7 , further comprising an agent. 
     
     
         9 . The composition of  claim 8 , wherein the agent is selected from the group consisting of:
 an antibiotic; an agent to attract cells; a cell; a stem cell; a ligand; a growth factor; a platelet; and a component of extracellular matrix.   
     
     
         10 . The composition of any of  claims 1 - 9 , wherein the average pore size of the biomaterial matrix is at least 200 μm. 
     
     
         11 . The composition of any of  claims 1 - 10 , wherein the average pore size of the biomaterial matrix is at least 300 μm. 
     
     
         12 . The composition of any of  claims 1 - 11 , wherein the average pore size of the biomaterial matrix is at least 400 μm. 
     
     
         13 . The composition of any of  claims 1 - 12 , wherein the average pore size of the biomaterial matrix is from about 200 μm to about 600 μm. 
     
     
         14 . The composition of any of  claims 1 - 13 , wherein the average pore size of the biomaterial matrix is from about 300 μm to about 500 μm. 
     
     
         15 . The composition of any of  claims 1 - 14 , wherein the average pore size of the biomaterial matrix is about 400 μm. 
     
     
         16 . The composition of any of  claims 1 - 15 , wherein the average pore size of the biomaterial matrix is about 300 μm. 
     
     
         17 . The composition of any of  claims 1 - 16 , wherein the impermeable layer is from about 10 μm to about 600 μm thick. 
     
     
         18 . The composition of any of  claims 1 - 17 , wherein the impermeable layer is from about 100 μm to about 400 μm thick. 
     
     
         19 . The composition of any of  claims 1 - 18 , wherein the impermeable layer is from about 150 μm to about 250 μm thick. 
     
     
         20 . The composition of any of  claims 1 - 19 , wherein the impermeable layer is from about 25 μm to about 150 μm thick. 
     
     
         21 . The composition of any of  claims 1 - 20 , wherein the impermeable layer is about 200 μm thick. 
     
     
         22 . The composition of any of  claims 1 - 21 , wherein the composition is from about 0.01 cm to about 5 cm thick. 
     
     
         23 . The composition of any of  claims 1 - 22 , wherein the composition is from about 0.1 cm to about 3 cm thick. 
     
     
         24 . The composition of any of  claims 1 - 23 , wherein the composition is from about 0.1 cm to about 2 cm thick. 
     
     
         25 . The composition of any of  claims 1 - 24 , wherein the composition is about 1 cm thick. 
     
     
         26 . The composition of any of  claims 1 - 25 , wherein the composition has a shape selected from the group consisting of:
 a sheet; a tube; and a contoured sheet.   
     
     
         27 . A method of producing a composition of any of  claims 1 - 26 , the method comprising;
 contacting a porous biomaterial matrix with a impermeable biomaterial layer.   
     
     
         28 . A method of producing a composition of any of  claims 1 - 26 , the method comprising;
 (c) casting an admixture of aqueous biomaterial solution and NaCl;   (d) contacting the composition resulting from step (α) with water.   
     
     
         29 . The method of  claim 28 , wherein step (α) is performed with the solution in contact with a pre-existing impermeable biomaterial layer. 
     
     
         30 . The method of any of  claims 28 - 29 , wherein the NaCl is granular. 
     
     
         31 . The method of any of  claims 28 - 30 , wherein the biomaterial solution is a silk fibroin solution. 
     
     
         32 . The method of any of  claims 28 - 31 , wherein the silk fibroin solution has a concentration of from about 2% wt/vol to about 15% wt/vol. 
     
     
         33 . The method of any of  claims 28 - 32 , wherein the silk fibroin solution has a concentration of from about 4% wt/vol to about 10% wt/vol. 
     
     
         34 . The method of any of  claims 28 - 33 , wherein the silk fibroin solution has a concentration of about 6% wt/vol. 
     
     
         35 . The method of any of  claims 28 - 34 , wherein the admixture is cast for from about 12 hours to about 96 hours. 
     
     
         36 . The method of any of  claims 28 - 35 , wherein the admixture is cast for from about 24 hours to 72 hours. 
     
     
         37 . The method of any of  claims 28 - 36 , wherein the admixture is cast for about 48 hours. 
     
     
         38 . The method of any of  claims 28 - 37 , wherein step (b) proceeds for from about 12 hours to about 120 hours. 
     
     
         39 . The method of any of  claims 28 - 38 , wherein step (b) proceeds for from about 24 hours to about 96 hours. 
     
     
         40 . The method of any of  claims 28 - 39 , wherein step (b) proceeds for about 72 hours. 
     
     
         41 . The method of any of  claims 28 - 40 , wherein the water is distilled water. 
     
     
         42 . The method of any of  claims 28 - 41 , wherein the water is removed and replaced with a fresh volume of water at least once during step (b). 
     
     
         43 . The method of any of  claims 28 - 42 , wherein, during the casting step, the admixture of biomaterial solution and NaCl is in contact with an impermeable layer. 
     
     
         44 . The method of any of  claims 27 - 43 , wherein the impermeable biomaterial layer comprises silk fibroin. 
     
     
         45 . The method of any of  claims 27 - 44 , wherein the method further comprises a first step of casting silk fibroin solution to form an impermeable silk fibroin layer. 
     
     
         46 . The method of  claim 45 , wherein the silk fibroin solution has a concentration of from about 2% wt/vol to about 15% wt/vol. 
     
     
         47 . The method of  claim 45 , wherein the silk fibroin solution has a concentration of from about 4% wt/vol to about 10% wt/vol. 
     
     
         48 . The method of  claim 45 , wherein the silk fibroin solution has a concentration of about 8% wt/vol. 
     
     
         49 . The method of any of  claims 27 - 48 , wherein the composition is produced under sterile conditions or sterilized after the rinsing step is complete. 
     
     
         50 . The method of any of  claims 27 - 49 , wherein the composition is produced in a linear shape. 
     
     
         51 . The method of any of  claims 27 - 50 , wherein the composition is produced in a tubular shape. 
     
     
         52 . The method of any of  claims 27 - 51 , wherein the method further comprises adding an agent to at least one layer. 
     
     
         53 . The method of  claim 52 , wherein the agent is a therapeutic agent. 
     
     
         54 . The method of any of  claims 27 - 53 , wherein the method further comprises altering the β-sheet content of a silk fibroin layer. 
     
     
         55 . The method of  claim 54 , wherein the β-sheet content of a silk fibroin layer is altered using a method selected from the group consisting of:
 contacting the layer with water vapor; drying; dehydration; water annealing; 
 stretching; compression; solvent immersion; immersion in methanol; 
 immersion in ethanol; pH adjustment; heat treatment; and sonication. 
 
     
     
         56 . The method of any of  claims 54 - 55 , wherein the β-sheet content of a silk fibroin layer is altered after adding an agent to the layer. 
     
     
         57 . A method for wound healing or repair of a tissue defect, the method comprising applying a composition of any of  claims 1 - 26  to the wound or tissue defect. 
     
     
         58 . The method of  claim 57 , wherein the wound or tissue defect is located in a hollow organ. 
     
     
         59 . The method of  claim 58 , wherein the hollow organ is selected from the group consisting of:
 the bladder; a portion of the gastrointestinal tract; the stomach, and the intestines.   
     
     
         60 . The method of  claim 57 , wherein the wound or tissue defect is selected from the group consisting of:
 a skin wound or defect; a diabetic ulcer; a bone wound or defect; a joint wound or defect; a meniscus wound or defect; an articular cartilage wound or defect; a soft tissue wound or defect; a lung tissue wound or defect; and a kidney wound or defect.   
     
     
         61 . The method of any of  claims 57 - 60 , wherein the impermeable layer is oriented to prevent the movement of material into or out of the wound or defect.