Hydrogel scaffolds for tissue engineering
View Patent ↗Disclosed herein are biodegradable hydrogel scaffolds for use in tissue engineering. The hydrogel scaffolds are composed of synthetic terpolymers complexed with polyvinyl alcohol (PVA), which facilitate cell-sheet and tissue growth. In the presence of a monosaccharide, the PVA-hydrogel is dissolved and cell-sheets are released for harvesting. Further disclosed herein are methods for producing PVA hydrogels which support tissue growth. Tissue engineering applications and methods are also disclosed.
1. A hydrogel scaffold composition comprising:
a hydrogel layer, the hydrogel layer comprising one or more polyvinyl alcohol polymers and one or more phenylboronate-containing copolymers, wherein the hydrogel layer is configured for cell growth into one or more of cell-sheets or tissues; and
a polyvinylidene fluoride membrane, wherein the polyvinylidene fluoride membrane overlays the hydrogel layer.
2. The composition of claim 1 , wherein the one more phenylboronate-containing copolymers includes one or more phenylboronate ligands, one or more acrylic monomers, and one or more alkaline tertiary amines.
3. The composition of claim 2 , wherein the one or more phenylboronate ligands are selected from the group consisting of 4-vinylphenylboronic acid, N-acryloyl-3-aminophenylboronic acid, 3-(2-carboxyvinyl)phenylboronic acid, 4-(1,6-dioxo-2,5-diaza-7-oxamyl)phenylboronic acid, and [2-(diethylamino)carbonyl-4-bromomethyl]phenylboronate, or any combination thereof.
4. The composition of claim 2 , wherein the one or more acrylic monomers are selected from the group consisting of acrylamide; N-isopropylacrylamide; N,N-dimethylacrylamide; 2-hydroxyethylmethacrylate; N-Hydroxyethyl acrylamide; methacrylamide; methacrylic acid; acrylic acid; N-vinyl-2-pyrrolidone; 4-pentenoic acid; N-isopropylmethacrylamide; N-methoxymethyl-N-isopropylacrylamide; and 2-(dimethylmaleimido)-N-ethylacrylamide, or any combination thereof.
5. The composition of claim 2 , wherein the one or more alkaline tertiary amines are selected from the group consisting of N,N-dimethylaminoethylmethacrylate; N,N-dimethylaminopropylacrylamide; N,N-diethylaminoethyl methacrylate; and 2-(N,N-Dimethylamino)ethyl methacrylate, or any combination thereof.
6. The composition of claim 1 , wherein the hydrogel scaffold is susceptible to degradation by saccharides.
7. The composition of claim 6 , wherein the saccharides are selected from the group consisting of fructose, glucose, mannitol, L-arabinose, D-xylose, D-galactose, D-mannose, L-fucose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and N-acetylneuraminic acid, or any combination thereof.
8. A method for producing the hydrogel scaffold of claim 1 comprising:
mixing one or more polyvinyl alcohol polymers with one or more phenylboronate-containing copolymers to form a mixture; and
allowing the mixture to congeal, wherein the scaffold is configured for cell growth into one or more of cell-sheets or tissues.
9. The method of claim 8 , wherein the one or more phenylboronate-containing copolymers includes one or more phenylboronate ligands, one or more acrylic monomers, and one or more alkaline tertiary amines.
10. The method of claim 8 , further comprising dissolving the one or more polyvinyl alcohol polymers prior to mixing the one or more polyvinyl alcohol polymers with the one or more phenylboronate-containing copolymers.
11. The method of claim 10 , wherein dissolving the one or more polyvinyl alcohol polymers prior to mixing the one or more polyvinyl alcohol polymers with the one or more phenylboronate-containing copolymers occurs at a temperature of at least 70° C.
12. The method of claim 11 , further comprising allowing the one or more polyvinyl alcohol polymers to cool to at least 40° C. after dissolving the one or more polyvinyl alcohol polymers and prior to mixing the one or more polyvinyl alcohol polymers with the one or more phenylboronate-containing copolymers.
13. The method of any of claim 8 , wherein mixing the one or more polyvinyl alcohol polymers with the one or more phenylboronate-containing copolymers to form the mixture occurs at a pH from 5 to 9.
14. A method for tissue engineering comprising:
culturing cells on the hydrogel scaffold of claim 1 to form one or more cell-layers, wherein the scaffold is configured for cell growth into one or more of cell-sheets or tissues;
dissolving the hydrogel scaffold with a saccharide solution; and
harvesting the one or more cell-layers.
15. The method of claim 14 , wherein the hydrogel scaffold includes one or more polyvinyl alcohol polymers.
16. The method of claim 14 , wherein the one or more phenylboronate-containing copolymers includes one or more phenylboronate ligands, one or more acrylic monomers, and one or more alkaline tertiary amines.
17. The method of claim 16 , wherein the one or more phenylboronate ligands are selected from the group consisting of 4-vinylphenylboronic acid, N-acryloyl-3-aminophenylboronic acid, 3-(2-carboxyvinyl)phenylboronic acid, 4-(1,6-dioxo-2,5-diaza-7-oxamyl)phenylboronic acid, and [2-(diethylamino)carbonyl-4-bromomethyl]phenylboronate, or any combination thereof.
18. The method of claim 16 , wherein the one or more acrylic monomers are selected from the group consisting of acrylamide; N-isopropylacrylamide; N,N-dimethylacrylamide; 2-hydroxyethylmethacrylate; N-Hydroxyethyl acrylamide; methacrylamide; methacrylic acid; acrylic acid; N-vinyl-2-pyrrolidone; 4-pentenoic acid; N-isopropylmethacrylamide; N-methoxymethyl-N-isopropylacrylamide; and 2-(dimethylmaleimido)-N-ethylacrylamide, or any combination thereof.
19. The method of claim 16 , wherein the one or more alkaline tertiary amines are selected from the group consisting of N,N-dimethylaminoethylmethacrylate; N,N-dimethylaminopropylacrylamide; N,N-diethylaminoethyl methacrylate; and 2-(N,N-Dimethylamino)ethyl methacrylate, or any combination thereof.
20. The method of claim 14 , wherein culturing the cells on the hydrogel scaffold occurs at a pH from 5 to 9.