Method for producing a collagen membrane and uses thereof

1. A method of producing a collagen-containing membrane comprising the steps of:

(i) isolating a collagen-containing tissue and incubating the isolated collagen-containing tissue in an ethanol solution;

(ii) incubating the isolated collagen-containing tissue from step (i) in a solution comprising about 1% (v/v) SDS and about 0.2% (v/v) LiCl in order to denature non-collagenous proteins contained therein;

(iii) incubating the collagen-containing tissue produced in step (ii) in a solution comprising about 0.5% (v/v) HCl until the collagen in said material is denatured; and

(iv) incubating the collagen-containing tissue produced in step (iii) in a solution comprising about 1% (v/v) HCl with simultaneous mechanical stimulation for sufficient time to enable the collagen bundles in said collagen-containing tissue to align; wherein the mechanical stimulation comprises applying tension cyclically to the collagen-containing tissue, thereby producing said collagen-containing membrane.

2. The method of claim 1 , wherein the ethanol solution comprises greater than 70% ethanol.

3. The method of claim 1 , wherein the incubation in step (ii) is at about 4° C.

4. The method of claim 1 , wherein the incubation in step (ii) is undertaken for at least 12 hours.

5. The method of claim 1 , wherein the incubation in step (iii) is undertaken for about 30 minutes.

6. The method of claim 1 , wherein the incubation in step (iii) is undertaken with shaking.

7. The method of claim 1 , wherein the incubation in step (iv) is undertaken for about 12 to 36 hours.

8. The method of claim 7 , wherein the incubation in step (iv) is undertaken for about 24 hours.

9. The method of claim 1 , wherein the incubation in step (iv) is undertaken with shaking.

10. The method of claim 1 , further comprising a neutralization step between step (iii) and step (iv) which comprises incubation of said collagen-containing tissue with about 0.5% (v/v) NaOH.

11. The method of claim 1 , further comprising step (v) which comprises incubating the collagen-containing tissue from step (iv) with acetone and then drying the collagen-containing tissue.

12. The method of claim 1 , wherein between steps (ii) and (iii) and/or between steps (iii) and (iv) the collagen-containing tissue is contacted with glycerol in order to visualise and facilitate the removal of fat and/or blood vessels.

13. The method of claim 12 , wherein the glycerol is contacted with the collagen-containing tissue for at least 10 minutes.

14. The method of claim 1 , wherein between steps (ii) and (iii) and/or between steps (iii) and (iv) the collagen-containing tissue is washed.

15. The method of claim 14 , wherein the wash used between steps (ii) and (iii) is acetone to remove the denatured proteins.

16. The method of claim 15 , wherein the collagen-containing tissue is further washed after the acetone with sterile water.

17. The method of claim 15 , wherein the collagen-containing tissue is further washed in a NaOH:NaCl solution.

18. The method of claim 1 , wherein after steps (iv) the collagen-containing tissue is washed with sterile water.

19. The method of claim 18 , wherein the collagen-containing tissue is further washed after the sterile water with the first solution.

20. The method of claim 1 , wherein the collagen-containing tissue comprises dense connective tissue.

21. The method of claim 1 , wherein the collagen-containing tissue is isolated from a sheep, a cow, a pig or a human.

22. The method of claim 1 , wherein the collagen-containing tissue is isolated from a human.

23. The method of claim 1 , wherein the collagen-containing tissue is autologous.