Ultra-thin film silk fibroin/collagen composite implant and manufacturing method therefor

Patent metadata

Field	Value
Publication number	US-10434216-B2
Application number	US-201515505054-A
Country	US
Kind code	B2
Filing date	Jul 10, 2015
Priority date	Aug 19, 2014
Publication date	Oct 8, 2019
Grant date	Oct 8, 2019

Abstract

The present invention relates to an ultra-thin film silk fibroin/collagen composite implant for tissue engineering and a manufacturing method therefor. The ultra-thin film silk fibroin/collagen silk fibroin/collagen composite implant according to the present invention has no cytotoxicity and can minimize the influence on cell growth, due to the combined use of a refined silk fibroin aqueous solution, collagen and various biomaterials, and thus can be widely used as an ultra-thin film implant for implanting.

First claim

The invention claimed is:

1. A method of manufacturing a silk fibroin-collagen composite film, the method comprising:
mixing a silk fibroin solution and a collagen solution to provide a liquid mixture comprising both silk fibroin and collagen;
applying, on a surface, the liquid mixture to provide a liquid layer comprising both silk fibroin and collagen;
subjecting the liquid layer to drying to provide a dried layer comprising both silk fibroin and collagen;
subjecting the dried layer to cross-linking of collagen therein to provide a cross-linked layer comprising both silk fibroin and cross-linked collagen; and
subsequently subjecting the cross-linked layer to crystallization of silk fibroin therein to provide a silk fibroin-collagen composite film comprising both silk fibroin crystals and cross-linked collagen.

2. The method of claim 1 , wherein subjecting collagen of the dried layer to a cross-linking reaction comprises contacting the dried layer with a solution of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, EDC/N-hydroxysuccinimide, or 2-chloro-1-methylpyridinium iodide in a concentration of 0.1 to 3%.

3. The method of claim 1 , wherein subjecting silk fibroin in the cross-linked layer to crystallization of silk fibroin comprises contacting the cross-linked layer with methanol or ethanol.

4. The method of claim 1 , further comprising:
subsequently to subjecting silk fibroin in the cross-linked layer to crystallization of silk fibroin, separating the silk fibroin-collagen composite film from the surface.

5. The method according to claim 1 , wherein the silk fibroin solution comprises 1 to 10 v/v % of sericin such that the liquid mixture further comprises sericin in addition to silk fibroin and collagen.

6. The method according to claim 1 , wherein the liquid mixture further comprises hyaluronic acid.

7. The method according to claim 1 , wherein the liquid mixture further comprises a biomaterial.

8. The method according to claim 1 , further comprising seeding corneal endothelium in the silk fibroin/collagen composite film.

9. A method for manufacturing a corneal endothelial transplantation implant comprising:
performing the method of claim 1 to provide a silk fibroin-collagen composite film comprising both silk fibroin crystals and cross-linked collagen therein;
seeding corneal endothelium on the silk fibroin-collagen composite film; and
culturing the corneal endothelium in the silk fibroin-collagen composite film to provide a corneal endothelial transplantation implant comprising the silk fibroin-collagen composite film and cultured corneal endothelium.

Assignees

• Nat Univ Chonbuk Ind Coop Found

Inventors

• Khang Gil Son
• Kim Eun Young
• Yang Jae Won
• Song Jeong Eun
• Cha Se Rom

Classifications

• A61L27/26 Primary

  Mixtures of macromolecular compounds · CPC title

• A61L2300/00

  Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices · CPC title

• A61L27/222

  Gelatin · CPC title

• A61L27/227

  Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24 · CPC title

• A61L27/20

  Polysaccharides · CPC title