Collagen fibrillar construction

Patent metadata

Field	Value
Publication number	US-9518106-B2
Application number	US-201514664137-A
Country	US
Kind code	B2
Filing date	Mar 20, 2015
Priority date	Apr 11, 2008
Publication date	Dec 13, 2016
Grant date	Dec 13, 2016

Abstract

Methods and compositions are described for organizing collagen into fibrillar networks, e.g, short and long-range organization. Collagen produced by the disclosed methods can be used for tissue engineering.

First claim

The invention claimed is:

1. A method of producing an organized array of collagen fibrils, the method comprising:
applying a tension to a plurality of collagen fibrils in a solution, wherein the tension is controlled to generate a strain of about 1% to about 10% such that a controlled load on collagen monomers in the solution is produced; and
adding a collagen lytic protease to the solution, thereby producing an organized array of collagen fibrils.

2. The method of claim 1 , further comprising neutralizing the pH of the solution.

3. The method of claim 2 , further comprising neutralizing the pH of the solution at about 10° C. to about 39° C.

4. The method of claim 1 , wherein the tension is applied to both ends of the plurality of collagen fibrils.

5. The method of claim 4 , further comprising adding supplemental collagen monomers to the solution continuously when applying the tension.

6. The method of claim 5 , wherein the collagen lytic protease and the supplemental collagen monomers are added simultaneously to the solution.

7. The method of claim 5 , wherein the collagen lytic protease and the supplemental collagen monomers are added sequentially to the solution.

8. The method of claim 5 , wherein the collagen lytic protease and the supplemental collagen are added more than once to the solution.

9. The method of claim 1 , further comprising organizing the array of collagen fibrils into a tissue.

10. The method of claim 5 , further comprising continuously extending a collagen fibrillar structure at a rate of from about 0.1 μm/min to about 100 μm/min.

11. The method of claim 10 , further comprising adding collagen lytic protease to the solution while extending the collagen fibrillar structure.

12. The method of claim 10 or 11 , further comprising extending the collagen fibrillar structure in the presence of a co-nonsolvency agent.

13. The method of claim 12 , wherein the co-nonsolvency agent is polyethylene glycol, hyaluronic acid, a glycosaminoglycan, a proteoglycan, or a combination thereof.

14. The method of claim 13 , wherein the proteoglycan is lumican, decorin, biglycan, perlecan, versican, fibromodulin, aggrecan, sydecan or a combination thereof.

15. The method of claim 1 or claim 11 , wherein the collagen lytic protease is a bacterial collagenase, a matrix metalloproteinase, or cathepsin.

16. The method of claim 1 , wherein the solution further comprises a collagen binding agent.

17. The method of claim 16 , wherein the collagen binding agent is a proteoglycan, a glycoprotein, a collagen-binding portion thereof, or a combination thereof.

18. The method of claim 1 , wherein the controlled load on each collagen monomer is from 1.0 piconewtons to 10.0 piconewtons.

19. The method of claim 1 , wherein the controlled load on each collagen monomer is greater than 10.0 piconewtons.

20. The method of claim 1 , further comprising adding a cross-linking agent to the solution.

21. The method of claim 20 , wherein the cross-linking agent is added before applying the tension.

22. The method of claim 20 , wherein the cross-linking agent is added after applying the tension.

23. The method of claim 20 , wherein the cross-linking agent is formaldehyde, hexamethylene diisocyanate, glutaraldehyde, polyepoxy compounds, gamma irradiation, or ultraviolet irradiation with riboflavin.

Assignees

• Univ Northeastern

Inventors

• Saeidi Nima
• Ruberti Jeffrey W

Classifications

• C08H1/06 Primary

  derived from horn, hoofs, hair, skin or leather · CPC title

• C08L89/06

  derived from leather or skin {, e.g. gelatin} · CPC title

• C07K14/78 Primary

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] · CPC title