Engineered tissue substitute system

Patent metadata

Field	Value
Publication number	US-10865811-B2
Application number	US-201615547320-A
Country	US
Kind code	B2
Filing date	Feb 3, 2016
Priority date	Feb 6, 2015
Publication date	Dec 15, 2020
Grant date	Dec 15, 2020

Abstract

Compositions and methods of preparation and use are provided for an engineered tissue substitute system comprising collagen, glycosaminoglycan and hydrogel in a cross-linked matrix. The compositions may be further lyophilized and reconstituted with a physiological fluid prior to use in methods, such as in the treatment of wounds, tissue engineering and cell transplantation.

First claim

What is claimed is:

1. A lyophilized powder composition, the composition comprising:
(a) collagen, wherein the collagen is at a concentration of between 3-10 mg/ml;
(b) glycosaminoglycan, wherein the ratio of the glycosaminoglycan to collagen is a weight ratio from about 4:1 to about 8:1;
(c) a biocompatible small molecule cross-linker of collagen and glycosaminoglycan;
(d) a hydrogel, wherein the hydrogel is 0.01%-1.0% w/vol of the final composition; and
(e) a biocompatible, small-molecule, hydrogel cross-linker.

2. The composition of claim 1 , wherein the biocompatible small molecule cross-linker of collagen and glycosaminoglycan is selected from one or more of the following: glutaraldehyde, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC), EDC (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide):NHS (N-hydroxysuccinimide), EDC (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide):sulfo-NHS (N-hydroxysulfoxuccinimide), hexamethylene diisocyanate and Genipin.

3. The composition of claim 1 , wherein the hydrogel is formed from hydrogel polymers, and wherein the hydrogel polymers are selected from one or more of: polyvinyl alcohol (PVA); polyvinyl acetate (PVacetate); thiolated polyvinyl alcohol; polyvinyl alcohol block polymers containing polyethylene glycol (PVA-PEG); polyvinylpyrrolidone (PVP); and a copolymer thereof of any two or more of the preceding polymers.

4. The composition of claim 1 , wherein the hydrogel is 0.01-0.5% w/vol of the final composition.

5. The composition of claim 1 , wherein the composition further comprises a solvent.

6. The composition of claim 5 , wherein the solvent is cell media, water, or a physiological fluid.

7. The composition of claim 6 , wherein the physiological fluid is blood, serum or plasma.

8. The composition of claim 1 , wherein fibrillogenesis begins within 13-16 minutes of solvent addition.

9. The composition of claim 1 , wherein gelation of the powder occurs:
between 25°-40° C.;
between 25°-37° C.;
between 30°-37° C.; or
at 37° C.

10. A wound treatment method, the method comprising reconstituting the powdered composition of claim 1 in a solvent and administration of the reconstituted composition to a patient in need thereof.

11. A method of preparing a composition of claim 1 , the method comprising:
(a) mixing collagen with glycosaminoglycan, wherein the ratio of the glycosaminoglycan to collagen is a weight ratio from about 4:1 to about 8:1;
(b) cross-linking of collagen and glycosaminoglycan;
(c) adding a hydrogel to the cross-linked collagen and glycosaminoglycan, wherein the hydrogel is 0.3%-1.0% w/vol of the final composition; and
(d) cross-linking the hydrogel.

12. The method of claim 11 , wherein the cross-linking of collagen and glycosaminoglycan is by dehydrothermal treatment (DHT), ultraviolet irradiation (UV) or enzymatic crosslinking.

13. The method of claim 11 , wherein the cross-linking of collagen and glycosaminoglycan is by small molecule cross-linker of collagen and glycosaminoglycan.

14. The method of claim 13 , wherein the cross-linker of collagen and glycosaminoglycan is selected from one or more of the following: glutaraldehyde, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC), EDC (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide):NHS (N-hydroxysuccinimide), EDC (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide):sulfo-NHS (N-hydroxysulfoxuccinimide), hexamethylene diisocyanate and Genipin.

15. The method of claim 14 , wherein the method further comprises adding dextran.

16. The method of claim 11 , wherein the hydrogel comprises one or more of: polyvinyl alcohol (PVA); polyvinyl acetate (PVacetate); thiolated polyvinyl alcohol; polyvinyl alcohol block polymers containing polyethylene glycol (PVA-PEG); polyvinylpyrrolidone (PVP); and a copolymer thereof of any two or more of the preceding polymers.

17. The method of claim 16 , wherein the hydrogel is a PVA hydrogel.

18. The method of claim 17 , wherein the PVA hydrogel is 0.4-0.6% w/vol of the final composition.

19. The method of claim 17 , wherein the cross-linker of the hydrogel is:
borate; or
sodium borate decahydrate.

20. The method of claim 11 , wherein the cross-linker is:
0.01%-0.0001% w/vol of the final composition; or
0.01-0.1% w/vol of the final composition.

21. The method of claim 11 , wherein the method further comprises lyophilizing of the composition.

22. The method of claim 21 , wherein the method further comprises powdering the lyophilized composition.

23. The method of claim 22 , wherein the method further comprises reconstituting the powder in a solvent.

24. The method of claim 23 , wherein the solvent is a physiological fluid or water.

25. The method of claim 24 , wherein the physiological fluid is blood, serum or plasma.

Assignees

• Univ British Columbia

Inventors

• Ghahary Aziz
• Hartwell Ryan

Classifications

• C12N2533/70

  Polysaccharides · CPC title

• A61L2300/64

  Animal cells · CPC title

• C12N2533/54

  Collagen; Gelatin · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• A61L27/48

  with macromolecular fillers · CPC title