Silk Fibroin Hydrogels and Uses Thereof
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.
1 . A method of soft tissue reconstruction or augmentation, the method comprising the step of administering a silk fibroin hydrogel formulation to a soft tissue region of an individual in need of soft tissue reconstruction or augmentation; wherein the formulation comprises,
a) a gel phase, the gel phase including hydrogel particles comprising a substantially sericin-depleted silk fibroin and an amphiphilic peptide; and
b) a carrier phase.
2 . The method of claim 1 , wherein the hydrogel particles comprises about 1% (w/v) to about 10% (w/v) of silk fibroin.
3 . The method of claim 1 , wherein the amphiphilic peptide comprising a RGD motif.
4 . The method of claim 3 , wherein the amphiphilic peptide is 23 RGD.
5 . The method of claim 1 , wherein the amphiphilic peptide comprises of a tail region, followed by a spacer region and finally a RGD motif.
6 . The method of claim 1 , wherein the hydrogel particles comprises a molar ratio of 1:10 to 10:1 moles of the amphiphilic peptide per mole of the silk fibroin.
7 . The method of claim 6 , wherein the hydrogel particles comprises a molar ratio of 3:1 moles of the amphiphilic peptide per mole of the silk fibroin.
8 . The method of claim 1 wherein the hydrogel particles comprises a protein structure having a β-sheet conformation of at least 80%.
9 . The method of claim 1 wherein the hydrogel particles comprises a protein structure having a β-sheet conformation of at least 50%.
10 . The method of claim 1 wherein the hydrogel particles comprises a protein structure having a β-sheet conformation of at least 20%.
11 . The method of claim 1 , wherein the hydrogel particles comprises a protein structure having an α-helical and random coil conformation of at most 20%.
12 . The method of claim 1 , wherein the hydrogel particles have a cross-sectional area from about 5 μm 2 to about 1000 μm 2 .
13 . The method of claim 1 , wherein the hydrogel particles have a cross-sectional area from about 20 μm 2 to about 50 μm 2 .
14 . The method of claim 1 , wherein the carrier phase comprises saline.
15 . The method of claim 1 , wherein the carrier phase comprises a surfactant solution.
16 . The method of claim 1 wherein the gel phase is about 50% to about 99% of the total formulation volume, the remainder being a carrier solution.
17 . The method of claim 16 , wherein the gel phase is about 75% of the total formulation volume, the remainder being a carrier solution.
18 . The method of claim 1 , wherein the hydrogel formulation further comprising lidocaine.
19 . The method of claim 1 , wherein a synthetic molecule having the formula: (molecule X) n -(spacer peptide) 0-300 -(five-amino-acid peptide tail) is conjugated to the silk fibroin.
20 . The method of claim 1 , wherein the hydrogel particles comprise a cellular component
21 . The method of claim 20 , wherein the cellular component comprises stem cells.
22 . The method of claim 20 , wherein the cellular component comprises adipose cells.
23 . The method of claim 20 , wherein the cellular component comprises stem cells and adipose cells.
24 . The method of claim 1 , wherein the soft tissue region is breast tissue.
25 . A method of soft tissue reconstruction, the method comprising the step of
a) placing an implantable medical device into an individual at the desired location;
b) expanding the device by putting a silk fibroin hydrogel formulation into the device, wherein the formulation comprises,
i) a gel phase, the gel phase including hydrogel particles comprising a substantially sericin-depleted silk fibroin and an amphiphilic peptide; and
ii) a carrier phase.
26 . A method of breast reconstruction or augmentation, the method comprising the step of subdermal administration of a silk fibroin hydrogel formulation to a breast of an individual in need of breast reconstruction or augmentation; wherein the formulation comprises,
a) a gel phase, the gel phase including hydrogel particles comprising a substantially sericin-depleted silk fibroin and an amphiphilic peptide; and
b) a carrier phase.