Treatment of bioprosthetic tissues to mitigate post implantation calcification

• US 7,579,381 B2
• Filed: 03/23/2006
• Issued: 08/25/2009
• Est. Priority Date: 03/25/2005
• Status: Active Grant

• Current Assignee: Edwards Lifesciences Corporation
• Original Assignee: Edwards Lifesciences Corporation
• Inventors: Dove, Jeffrey S.
• Primary Examiner(s): Henley, III; Raymond J
• Application Number:
  US11/387,615
• Publication Number:
  US 20060217805A1
• Time in Patent Office: 1,251 Days
• Field of Search: 514/693, 514/694, 424/422
• US Class Current: 514/693
• CPC Class Codes: A61L 2400/02 Treatment of implants to prevent calcification or mineralisation in vivo
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment

Abstract

The present invention provides methods for treating tissue to inhibit post-implant calcification of a biological tissue. In one method of this invention, a tissue is immersed in or otherwise contacted with a pretreated glutaraldehyde solution, i.e., a heat-treated or pH-adjusted glutaraldehyde solution. The tissue may be partially fixed with glutaraldehyde prior to, after, or concurrently with the step of contacting the tissue with the pretreated gluteraldehyde. Contact with the pretreated gluteraldehyde produces free amine groups on the tissue, which are subsequently blocked by contacting the crosslinked tissue with a blocking agent. In another embodiment, a tissue is contacted with either a non-pretreated glutaraldehyde or a pH-adjusted glutaraldehyde solution for a period of time sufficient to crosslink the tissue. The crosslinked tissue is then treated with a reducing agent that reduces aldehyde and carboxylic acid groups on the fixed tissue.

Claims

1. A method for mitigating post-implantation calcification of a bioprosthetic material, said method comprising

(a) adjusting the pH of a glutaraldehyde solution to a pH in the range of about 5.0 to 7.0;

(b) contacting a quantity of biological tissue that contains connective tissue protein with the glutaraldehyde solution;

(c) heating the glutaraldehyde solution and the tissue to a temperature in the range of about 30 to 70°

C. for a period of time, thereby producing free amine groups on the tissue, wherein the tissue is

(i) at least partially fixed prior to the performance of Step (c),

(ii) fixed after the performance of Step (c), or

(iii) fixed concurrently with the performance of Step (c),

and wherein said tissue is fixed by immersing the tissue in a crosslinking solution comprising gluteraldehyde as a crosslinking agent; and

(d) contacting the fixed tissue with a solution containing a blocking reagent that blocks the free amine groups.

2. The method of claim 1, wherein the pH is adjusted to about 6.0.

3. The method of claim 1, wherein the temperature is in the range of about 40 to 60°

C.

4. The method of claim 1, wherein the period of time is between about one hour and six months.

5. The method of claim 1, wherein said blocking agent has a functional group that is reactive with the free amine groups.

6. The method of claim 1, wherein said blocking agent is a monofunctional aldehyde, a polyepoxy, a sugar, or collagen.

7. The method of claim 6, wherein said monofunctional aldehyde is formaldehyde.

8. The method of claim 6, wherein said polyepoxy is ethylene glycol diglycidyl ether.

9. The method of claim 1, wherein said glutaraldehyde has a low acid content.

10. The method of claim 1, wherein said crosslinking solution includes a non-isotonic buffer.

11. The method of claim 10, wherein said non-isotonic buffer is a hypotonic buffer.

12. The method of claim 1, further comprising contacting the tissue with a surfactant or a high osmolality aqueous solution of a salt and a sugar prior to Step (a).

13. The method of claim 1, further comprising subjecting the tissue to a bioburden reduction.

14. The method of claim 13, wherein the bioburden reduction process comprises contacting the tissue with a bioburden reduction solution containing a surfactant, and aldehyde and an alcohol.

15. The method of claim 1, further comprising sterilizing the bioprosthesis.

16. The method of claim 15, wherein the sterilization comprises contacting the bioprosthesis with a terminal sterilization solution and heating said terminal sterilization solution to a temperature between about 20 to 50°

C. for a period of time sufficient to ensure the sterility of the bioprosthesis until the time of implantation.

17. The method of claim 16, wherein said terminal sterilization solution comprises an aqueous solution of 0.2-1.0% by weight glutaraldehyde buffered to a pH of approximately 7.4.

18. The method of claim 16, wherein the terminal sterilization solution comprises osmotically balanced salt solution in combination with at least one chemical sterilant.

19. The method of claim 1, further comprising storing the bioprosthetic in a storage solution comprising gluteraldehyde and an antioxidant.

20. The method of claim 19, wherein said antioxidant is ascorbic acid.

21. The method of claim 1, wherein one or more of said steps is performed under non-oxidizing conditions.

22. The method of claim 21, wherein said non-oxidizing conditions are selected from the group consisting of;

a nitrogen blanket;

low actinic safety lights; and

mechanical covers.