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Last Updated: March 28, 2024

Claims for Patent: 9,096,688


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Summary for Patent: 9,096,688
Title:Method for producing a collagen membrane and uses thereof
Abstract: The present invention relates to a method of producing a collagen membrane that has particular mechanical properties. In particular, the present invention relates to a method A of producing a collagen membrane comprising the steps of (i) isolating a collagen-containing tissue and incubating same in an ethanol solution; (ii) incubating the collagen-containing tissue from step (i) in a first solution comprising an inorganic salt and an anionic surfactant in order to denature non-collagenous proteins contained therein; (iii) incubating the collagen-containing tissue produced in step (ii) in a second solution comprising an inorganic acid until the collagen in said material is denatured; and (iv) incubating the collagen-containing tissue produced in step (iii) in a third solution comprising an inorganic acid 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.
Inventor(s): Zheng; Ming Hao (City Beach, AU)
Assignee: Orthocell Pty Ltd. (AU)
Application Number:14/219,555
Patent Claims: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.degree. 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.

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