Silk Fibroin Hydrogels and Uses Thereof

a technology of silk fibroblasts and hydrogels, which is applied in the field of silk fibroblast hydrogels, can solve the problems of limiting the capacity of native host cells to bind to and interact with implanted silk devices, adverse tissue reactions, and heightened immune responses, so as to improve the utility and biocompatibility of silk formulations, improve the utility and function of silk formulations, and improve the function of medical devices

Inactive Publication Date: 2011-01-13
ALLERGAN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0012]The embodiments described herein provide for silk hydrogel formulations that may be useful for a variety of medical uses. More specifically, example embodiments of the present invention provide for gels including silk fibroin and peptides. Other example embodiments provide for the use of organic enhancers which improve device utility and functional peptide enhancers that may improve utility and biocompatibility of silk formulations. Silk hydrogel embodiments may be used as tissue space fillers, templates for tissue reconstruction or regeneration, cell culture scaffolds for tissue engineering and for disease models, surface coating to improve medical device function, or drug delivery devices.

Problems solved by technology

Sericin is often antigenic and may be associated with an adverse tissue reaction when sericin-containing silk is implanted in vivo.
Residual sericin that may be contained in these materials stands as a potential obstacle to its use as a biomaterial as it does present the possibility for a heightened immune response.
These two factors couple to severely limit the capacity of native host cells to bind to and interact with implanted silk devices, as neither inflammatory cells like macrophages or reparative cells like fibroblasts are able to attach strongly, infiltrate and bioresorb the silk fibroin devices.
This severely limits the subsequent long-term potential of the device particularly relating to tissue in-growth and remodeling and potentially, the overall utility of the device.
Increasing the silk concentration and temperature may alter the time taken for silk gelation by increasing the frequency of molecular interactions, increasing the chances of polymer nucleation.
The existence of sericin molecules in the silk solution intermediate prior to gelation may also compromise final gel structural quality, i.e., the distribution of β-sheet structure.

Method used

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  • Silk Fibroin Hydrogels and Uses Thereof
  • Silk Fibroin Hydrogels and Uses Thereof
  • Silk Fibroin Hydrogels and Uses Thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Silk Sericin Extraction

[0236]Silk fibroin for generation of the hydrogel was obtained in the form of degummed B. mori silk at a size of 20 denier-22 denier (38 μm±5.6 μm diameter). This degummed silk was further processed in order to remove the inherently present and potentially antigenic protein glue, sericin that conjoins independent fibroin filaments. This was done as described previously herein. Following removal of sericin, the pure fibroin was dried carefully to ambient humidity levels using a laminar flow hood.

example 2

Generation of Silk Fibroin Solution

[0237]Silk fibroin filaments, cleaned of their sericin and rinsed free of insoluble debris and ionic contaminants were used for the generation of an aqueous silk solution. These silk fibers were added to a solution of 9.3M LiBr and purified water (e.g., MILLI-Q® Ultrapure Water Purification Systems) (Millipore, Billerica, Mass.) to make a solution consisting of 20% pure silk (% w / v). This mixture was then heated to a temperature of 60° C. and digested for a period of four hours. A total of 12 mL of the resultant solution was then loaded into a 3 mL-12 mL Slide-A-Lyzer dialysis cassette (Pierce Biotechnology, Inc., Rockford, Ill.) (molecular weight cutoff of 3.5 kD) and placed into a beaker containing purified water as a dialysis buffer at a volume of 1 L water per 12 mL cassette of silk solution. The beakers were placed on stir plates and stirred continuously for the duration of the dialysis. Changes of dialysis buffer occurred at 1, 4, 12, 24, and...

example 3

Induction of Gelation

[0239]A variety of different methods were employed in the course of hydrogel development for the purposes of contrasting and comparing certain relevant properties of various formulae. Regardless of the nature in which the gelation process was carried out, the final determination that a “gel” state had been reached was applied uniformly to all groups. A solution or composite of solutions (i.e., silk solution blended with an enhancer or enhancer solution) was considered a gel after observing formation of a uniform solid phase throughout the entire volume, generally opaque and white in appearance.

[0240]Samples to be produced by passive gelation were not exposed to any enhancer additives. These gels were produced by measuring a volume of silk solution into a casting vessel, for the purposes of these experiments, polypropylene tubes sealed against air penetration and water loss, and the sample allowed to stand under ambient room conditions (nominally 20-24° C., 1 atm...

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Abstract

The present specification provides for methods for purifying fibroins, purified fibroins, methods of conjugating biological and synthetic molecules to fibroins, fibroins conjugated to such molecules, methods of making fibroin hydrogels, fibroin hydrogels and fibroin hydrogel formulations useful for a variety of medical uses, including, without limitation uses as bulking agents, tissue space fillers, templates for tissue reconstruction or regeneration, cell culture scaffolds for tissue engineering and for disease models, surface coating to improve medical device function, or drug delivery devices.

Description

CROSS REFERENCE[0001]This patent application claims priority pursuant to 35 U.S.C. §119(e) to U.S. Provisional Patent Application Ser. No. 61 / 170,895 filed Apr. 20, 2009, which is hereby incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]The present specification discloses purified silk fibroin and method for purifying silk fibroins, hydrogels comprising silk fibroin with or without an amphiphilic peptide and methods for making hydrogels comprising silk fibroin and the use of silk fibroin hydrogels in a variety of medical uses, including, without limitation fillers for tissue space, templates for tissue reconstruction or regeneration, scaffolds for cells in tissue engineering applications and for disease models, a surface coating to improve medical device function, or as a platform for drug delivery.BACKGROUND[0003]Silk refers to a filamentous product secreted by an organism such as a spider or silkworm. Fibroin is the primary structural component of silk. It is c...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61F2/10A61K31/167A61K35/12A61P9/06A61P23/02A61P41/00A61F2/52
CPCA61K38/1767A61K47/42A61L27/227A61L27/26A61L27/52A61K2800/91C08B37/0072A61F2/12A61K8/64A61Q19/00A61K2800/5922A61L2430/34C08L89/00A61P17/00A61P23/02A61P41/00A61P43/00A61P9/06
Inventor ALTMAN, GREGORY H.HORAN, REBECCA L.COLLETTE, ADAM L.CHEN, JINGSONGVANEPPS, DENNIS
Owner ALLERGAN INC
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