Treated tissue for implantation and methods of preparation

• US 5,632,778 A
• Filed: 06/05/1995
• Issued: 05/27/1997
• Est. Priority Date: 03/14/1994
• Status: Expired due to Term

• Current Assignee: Cryolife Incorporated
• Original Assignee: Cryolife Incorporated
• Inventors: Goldstein, Steven
• Primary Examiner(s): Prebilic, Paul B.
• Application Number:
  US08/463,643
• Time in Patent Office: 722 Days
• Field of Search: 623/11, 623/66, 128/898, 435/240.241, 435/240.243, 8/94.11, 8/94.18
• US Class Current: 424/423
• CPC Class Codes: A61L 2430/40 Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
• 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 27/3625 Vascular tissue, e.g. heart valves
• 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
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: Y10S 623/92 Method or apparatus for preparing or treating prosthetic
• CPC Class Codes: Y10S 623/921 Blood vessel

Abstract

This disclosure includes a method for generating a functional hybrid bioprosthesis. Tissue formed naturally of interstitial collagens is treated to kill native cells and remove potentially immunologically active soluble molecules. Then it may be treated sequentially with extracellular matrix adhesion factor, extracellular matrix glycosaminoglycan, and growth factor appropriate to the cell type required to function within the matrix, and incubating the transplant tissue matrix with cells that are either allogeneic or autologous for the recipient thereby imparting to the matrix the characteristics of the cell type and tissue selected. Tissues with a variety of functional bioactivities can thus be formed in vitro prior to graft transplantation or implantation which will exhibit reduced or no stimulation of an immunological response in the recipient.

Claims

1. A process for generating a substantially non-immunogenic tissue matrix suitable for subsequent processing into an implant tissue comprising:

A. eliminating native cells by treating a tissue with components selected from the group consisting enzymes and nucleases effective to inhibit subsequent native cell growth in the treated tissue and effective to limit generation of new immunological sites in the tissue thus producing a tissue matrix;

B. treating the tissue matrix with cellular adhesion factor to promote subsequent attachment of cultured allogeneic or autologous cells to the surfaces of the tissue matrix; and

C. repopulating the tissue matrix throughout the matrix with cultured allogeneic or autologous cells.

2. The process of claim 1 wherein, during step A, the tissue is treated with a low ionic strength solution of nucleases effective to decellularize the tissue and provide a tissue matrix of limited immunogenicity.

3. The process of claim 2 wherein the low ionic strength solution of nucleases includes at least one member, selected from the group consisting of RNAase A, DNAase I, EcoR I and Hind III.

4. The process of claim 2 wherein the nucleases employed are ribonuclease A and deoxyribonuclease I.

5. The process of claim 1 further comprising cryopreserving the tissue matrix after step A and prior to further processing.

6. The process of claim 2 further comprising cyropreserving the tissue matrix after step A and prior to further processing.

7. The process of claim 5 further comprising sterilizing the tissue or tissue matrix or both.

8. The process of claim 1 wherein the process further comprises obtaining natural mammalian tissue, cryopreserving the tissue, removing the tissue from cryopreservation and then treating the tissue in accordance with claim 1.

9. The process of claim 8 wherein the tissue is of porcine heart valve origin.

10. The process of claim 9 wherein the heart valve is a pulmonary or an aortic heart valve.

11. The process of claim 1 wherein step A further comprises treatment with a hypotonic aqueous solution.

12. The process of claim 11 herein said hypotonic aqueous solution is a buffered aqueous solution effective to cause cell lysis.

13. The process of claim 12 wherein the tissue is porcine heart valve tissue.

14. The process of claim 13 wherein the heart valve is a pulmonary or an aortic heart valve.

15. The process of claim 14 further comprising the step of cryopreserving the porcine heart valve tissue matrix produced by step A.