In-vitro method for the production of a homologous stented tissue-engineered heart valve

• US 20060246584A1
• Filed: 09/04/2002
• Published: 11/02/2006
• Est. Priority Date: 08/01/2002
• Status: Abandoned Application

• Current Assignee: University of Zurich
• Original Assignee: University of Zurich
• Inventors: Covelli, Bruno
• Application Number:
  US10/523,618
• Publication Number:
  US 20060246584A1
• Time in Patent Office: Days
• US Class Current: 435/396
• CPC Class Codes: A61F 2/2415 Manufacturing methods
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• 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 27/3645 Connective tissue
• 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 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3808 Endothelial cells
• CPC Class Codes: A61L 27/3843 Connective tissue
• 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: A61L 31/005 Ingredients of undetermined constitution or reaction products thereof
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence

Abstract

The invention relates to an in-vitro method for the production of a homologous stented tissue-engineered heart valve.

Claims

1. An in vitro method for the production of a homologous heart valve, comprising the steps of:

a) providing a biodegradable support,

b) colonizing the support with homologous fibroblasts or myofibroblasts cells or a combination thereof to form a connective tissue matrix,

c) optionally colonizing the connective tissue matrix with endothelial cells, and

d) fixing the connective tissue matrix to a non-degradable or poorly degradable frame construction,

wherein, before or after the fixing of the frame construction, the connective tissue matrix optionally colonized with endothelial cells is introduced into a pulsatile flow chamber in which it can be exposed to increasing flow rates, and the flow rate is increased continuously or discontinuously.

3. The method according to claims 1 or 2, wherein the biodegradable support comprises a biodegradable polymer matrix or an acellular biological matrix.

4. The method of claim 3, wherein the support comprises a polyglycolic acid (PGA), polylactic acid (PLA), polyhydroxyalkanoate (PHA), poly-4-hydroxybutyrate (P4HB) or a mixture of two or more of these polymers.

5. The method according to claims 1 or 2, wherein the support has a polymer density of 40 to 120 mg/cm³.

6. The method according to claims 1 or 2, wherein the support comprises a porous polymer having a pore size of 80 to 240 μ

m.

7. The method according to claims 1 or 2, wherein the fibers of the support have a diameter of 6 to 20 μ

m.

8. The method of claim 3, wherein the support comprises an acellular connective tissue framework of an animal or human heart valve.

9. The method according to claims 1 or 2, wherein the step of colonization with fibroblast or myofibroblast cells or a combination thereof repeated 3 to 14 times.

10. The method according to claims 1 or 2, wherein approximately 10⁵ to 6×

10⁸ fibroblast or myofibroblasts cells or a combination thereof are employed per square centimeter of support.

11. The method according to claims 1 or 2, wherein the step of colonization with endothelial cells is repeated 3 to 14 times.

12. The method according to claims 1 or 2, wherein approximately 10⁵ to 5×

10⁸ endothelial cells are employed per square centimeter of support.

13. The method according to claims 1 or 2, wherein the cells are human cells.

14. The method according to claims 1 or 2, wherein the cells are autologous cells.

15. The method according to claims 1 or 2, wherein the frame construction comprises a biocompatible material.

17. The method according to claims 1 or 2, wherein the support is fixed to the frame construction by means of conventional suturing, fibrin adhesive, or a combination thereof.

18. The method according to claim 1 or 2, wherein flow rates of 5 ml/min to 8,000 ml/min are established in the pulsatile flow chamber.

19. The method according to claims 1 or 2, wherein the flow rate is increased over a period of 1 week to 12 weeks.

20. The method according to claims 1 or 2, wherein the initial flow rate is 50 to 100 ml/min.

21. The method according to claims 1 or 2, wherein the initial pulse frequency is 5 to 10 pulses/min.

22. The method according to claims 1 or 2, wherein the flow rate is increased to 5,000 ml/min.

23. The method according to claims 1 or 2, wherein the pulse frequency is increased to 180 pulses/min.

24. The method according to claims 1 or 2, wherein systemic pressures of 10 to 240 mm Hg are established in the pulsatile flow chamber.

25. An autologous heart valve that has been produced by the method according to claims 1 or 2.

2. An in vitro method for the production of a homologous heart valve, comprising the following steps:

a) providing a biodegradable support which is firmly connected to a non-degradable or poorly degradable frame construction

b) colonizing the support with homologous fibroblast or myofibroblasts cells or a combination thereof to form a connective tissue matrix,

c) optionally colonizing the connective tissue matrix with endothelial cells,

d) introducing the frame construction with the connective tissue matrix connected thereto into a pulsatile flow chamber in which it can be exposed to increasing flow rates, and

e) continuously or discontinuously increasing of the flow rate.

16. (canceled)

26. An autologous heart valve having a connective tissue inner structure surrounded by an endothelial cell layer, wherein the connective tissue inner structure is fixed to a non-degradable or slowly degradable frame constructions.

27. The autologous heart valve according to claim 26, wherein a collagen density of 20 to 60% exists in the connective tissue inner structure.

28. The autologous heart valve according to claim 27, wherein the heart valve withstands the flow conditions in the human heart.