Silk fibroin fiber bundles for matrices in tissue engineering

Abstract

The present invention provides a novel silk-fiber-based matrix having a wire-rope geometry for use in producing a ligament or tendon, particularly an anterior cruciate ligament, ex vivo for implantation into a recipient in need thereof. The invention further provides the novel silk-fiber-based matrix which is seeded with pluripotent cells that proliferate and differentiate on the matrix to form a ligament or tendon ex vivo. Also disclosed is a bioengineered ligament comprising the silk-fiber-based matrix seeded with pluripotent cells that proliferate and differentiate on the matrix to form the ligament or tendon. A method for producing a ligament or tendon ex vivo comprising the novel silk-fiber-based matrix is also disclosed.

1 . A method for producing a sericin-extracted silk-fibroin fiber construct comprising:
 contacting at least one silkworm fibroin fiber with an aqueous solution of Na 2 Co 3  and detergent to extract sericin from the fiber.   
     
     
         2 . The method of  claim 1 , wherein the sericin is extracted at a temperature no greater than about 90° C. 
     
     
         3 . The method of  claim 1 , further comprising coating the fibroin fiber with a coating, a surface modifier, or a combination thereof that promotes cellular attachment on the fiber, tissue proliferation on the fiber, or a combination of cellular attachment and tissue proliferation on the fiber. 
     
     
         4 . The method of  claim 3 , wherein the coating, surface modifier, or combination thereof comprises an arginine-glycine-aspartate (RGD) peptide. 
     
     
         5 . The method of  claim 3 , wherein the coating, surface modifier, or combination thereof comprises a growth factor. 
     
     
         6 . The method of  claim 1 , further comprising placing the fiber in a twisted, helical, braided, mesh-like or cabled geometry. 
     
     
         7 . The method of  claim 1 , further comprising:
 contacting the construct with cells; and   culturing the construct under conditions suitable for cell growth and regeneration.   
     
     
         8 . A method for producing a sericin-extracted silk-fibroin fiber construct comprising:
 arranging at least two silkworm fibroin fibers to form a group; and   contacting the group with an aqueous solution of Na 2 Co 3  and detergent to extract sericin from the fibers.   
     
     
         9 . The method of  claim 8 , wherein the sericin is extracted at a temperature no greater than about 90° C. 
     
     
         10 . The method of  claim 8 , further comprising coating the fibroin fibers with a coating, a surface modifier, or a combination thereof that promotes cellular attachment on the fibers, tissue proliferation on the fibers, or a combination of cellular attachment and tissue proliferation on the fibers. 
     
     
         11 . The method of  claim 10 , wherein the coating, surface modifier, or combination thereof comprises an arginine-glycine-aspartate (RGD) peptide. 
     
     
         12 . The method of  claim 10 , wherein the coating, surface modifier, or combination thereof comprises a growth factor. 
     
     
         13 . The method of  claim 8 , wherein the group comprises up to 1300 fibroin fibers. 
     
     
         14 . The method of  claim 8 , wherein the fibers are organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry. 
     
     
         15 . The method of  claim 8 , further comprising:
 contacting the construct with cells; and   culturing the construct under conditions suitable for cell growth and regeneration.   
     
     
         16 . The method of  claim 15 , wherein the cells are stem cells, muscle cells, bone marrow stromal cells, pluripotent cells, or fibroblast cells. 
     
     
         17 . A method for producing a sericin-extracted silk-fibroin fiber construct comprising:
 arranging at least two groups of silkworm fibroin fibers to form a bundle; and   contacting the bundle with an aqueous solution of Na 2 Co 3  and detergent to extract sericin from the fibers.   
     
     
         18 . The method of  claim 17 , wherein the sericin is extracted at a temperature no greater than about 90° C. 
     
     
         19 . The method of  claim 17 , further comprising coating the fibroin fibers with a coating, surface modifier, or combination thereof that promotes cellular attachment on the fibers, tissue proliferation on the fibers, or a combination of cellular attachment and tissue proliferation on the fibers. 
     
     
         20 . The method of  claim 19 , wherein the coating, surface modifier, or combination thereof comprises an arginine-glycine-aspartate (RGD) peptide. 
     
     
         21 . The method of  claim 19 , wherein the coating, surface modifier, or combination thereof comprises a growth factor. 
     
     
         22 . The method of  claim 17 , wherein the groups are organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry forming a bundle. 
     
     
         23 . The method of  claim 17 , further comprising:
 contacting the construct with cells; and   culturing the construct under conditions suitable for cell growth and regeneration.   
     
     
         24 . The method of  claim 23 , wherein the cells are stem cells, muscle cells, bone marrow stromal cells, pluripotent cells, or fibroblast cells. 
     
     
         25 . A method for producing a sericin-extracted silk-fibroin fiber construct comprising:
 arranging at least two bundles of silkworm fibroin fibers to form a strand; and
 contacting the strand with an aqueous solution of Na 2 Co 3  and detergent to extract sericin from the fibers. 
   
     
     
         26 . The method of  claim 25 , wherein the sericin is extracted at a temperature no greater than about 90° C. 
     
     
         27 . The method of  claim 25 , further comprising coating the fibroin fibers with a coating, a surface modifier, or a combination thereof that promotes cellular attachment on the fibers, tissue proliferation on the fibers, or a combination of cellular attachment and tissue proliferation on the fibers. 
     
     
         28 . The method of  claim 27 , wherein the coating, surface modifier, or combination thereof comprises an arginine-glycine-aspartate (RGD) peptide. 
     
     
         29 . The method of  claim 27 , wherein the coating, surface modifier, or combination thereof comprises a growth factor. 
     
     
         30 . The method of  claim 25 , wherein the bundles are organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry forming a strand. 
     
     
         31 . The method of  claim 25 , further comprising:
 contacting the construct with cells; and   culturing the construct under conditions suitable for cell growth and regeneration.   
     
     
         32 . The method of  claim 31 , wherein the cells are stem cells, muscle cells, bone marrow stromal cells, pluripotent cells, or fibroblast cells. 
     
     
         33 . A method for producing a sericin-extracted silk-fibroin fiber construct comprising:
 arranging at least two strands of silkworm fibroin fibers to form a cord; and   contacting the cord with an aqueous solution of Na 2 Co 3  and detergent to extract sericin from the fibers.   
     
     
         34 . The method of  claim 33 , wherein the sericin is extracted at a temperature no greater than about 90° C. 
     
     
         35 . The method of  claim 33 , further comprising coating the fibroin fibers with a coating, a surface modifier, or a combination thereof that promotes cellular attachment on the fibers, tissue proliferation on the fibers, or a combination of cellular attachment and tissue proliferation on the fibers. 
     
     
         36 . The method of  claim 35 , wherein the coating, surface modifier, or combination thereof comprises an arginine-glycine-aspartate (RGD) peptide. 
     
     
         37 . The method of  claim 35 , wherein the coating, surface modifier, or combination thereof comprises a growth factor. 
     
     
         38 . The method of  claim 33 , wherein the strands are organized in a in parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry forming a cord. 
     
     
         39 . The method of  claim 33 , further comprising:
 contacting the construct with cells; and   culturing the construct under conditions suitable for cell growth and regeneration.   
     
     
         40 . The method of  claim 39 , wherein the cells are stem cells, muscle cells, bone marrow stromal cells, pluripotent cells, or fibroblast cells.