Production of tissue engineered heart valves

• US 20060253192A1
• Filed: 03/10/2006
• Published: 11/09/2006
• Est. Priority Date: 03/11/2005
• Status: Active Grant

• Current Assignee: Wake Forest University Health Sciences
• Original Assignee: Wake Forest University Health Sciences
• Inventors: Atala, Anthony, Yoo, James J.
• Granted Patent: US 9,248,015 B2
• Time in Patent Office: Days
• US Class Current: 623/2.130
• CPC Class Codes: A61F 2/2415 Manufacturing methods
• 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 27/3641 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3808 Endothelial cells
• CPC Class Codes: A61L 27/3826 Muscle cells, e.g. smooth muscle cells
• CPC Class Codes: A61L 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3886 comprising two or more cell types
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/069 Vascular Endothelial cells

Abstract

The invention is directed to methods for preparing artificial heart valves by preconditioning a matrix seeded with endothelial cells and smooth muscle cells differentiated from isolated progenitor cells. These cell seeded matrices are exposed to fluid conditions that mimic blood flow through the heart to produce tissue engineered heart valves that are analogous to native heart valves.

Claims

1. A method for producing a preconditioned heart valve, comprising:

obtaining a population of endothelial cells differentiated from progenitor cells;

seeding a matrix having a heart valve shape with the endothelial cells such the cells attach to the matrix to form an endothelial cell layer;

obtaining a population of smooth muscle cells differentiated from progenitor cells;

depositing the smooth muscle cells onto the endothelial cell layer such that the smooth muscle cells form a smooth muscle cell layer that joins to the endothelial cell layer;

attaching seeded matrix to an attachment element in a preconditioning chamber, wherein the attachment element has a channel that is fluidly coupled to fluid flow system; and

preconditioning the seeded matrix with the flow system in which a biological fluid is moved through the seeded matrix, wherein the flow-rate and pulse-rate of the biological fluid is controlled such that a preconditioned heart valve is produced.

2. The method of claim 1, wherein biocompatible matrix is selected from the group consisting of a decellularized matrix, an electrospun matrix, and a synthetic polymer matrix.

3. The method of claim 1, wherein the preconditioning chamber is a container with dimensions suitable for attaching heart valve leaflets.

4. The method of claim 1, wherein the step of preconditioning the matrix comprises moving the biological fluid through the inside surface of the attached matrix in a closed unidirectional fluid flow system.

5. The method of claim 1, wherein the step of preconditioning the matrix comprises moving the biological fluid through the inside surface of the attached matrix as a continuous flow.

6. The method of claim 5, wherein the continuous flow has a flow-rate that is incremented over time to induce a wall shear in the range of about 1 dyne/cm² to about 30 dynes/cm².

7. The method of claim 1, wherein the step of preconditioning the matrix comprises moving the biological fluid through the inside surface of the attached matrix as a pulsed flow.

8. The method of claim 7, wherein the pulsed flow has a pulse-rate that is incremented over time to induce a wall shear in the range of about 10 dynes/cm² to about 45 dynes/cm².

9. The method of claim 7, wherein the pulsed flow has a pulse-rate that is incremented over time to induce a wall pressure distribution in the range of about 60 to about 200 mmHg.

10. The method of claim 1, wherein the biological fluid is moved through the seeded tubular matrix by a pump.

11. The method of claim 1, wherein the biological fluid has a composition and viscosity that is equivalent to blood.

12. The method of claim 1, wherein the biological fluid is selected from the group consisting of culture medium, buffer medium, and physiological medium.

13. The method of claim 1, further comprising adding a volume of biological fluid to the preconditioning chamber such that the outside surface of the tubular matrix is exposed to the biological fluid.

14. The method of claim 1, wherein the matrix is an electrospun matrix comprising at least one natural component and at least one synthetic polymer component and a therapeutic agent coupled to a nanoparticle.

15. The method of claim 14, wherein the natural component is collagen and the synthetic polymer component is poly(lactide-co-glycolides) (PLGA).

16. The method of claim 14 further comprising elastin.

17. The method of claim 14, wherein the therapeutic agent is heparin and the nanoparticle is a quantum dot.

18. The method of claim 17, wherein the heparin and quantum dot are encapsulated in a polymer.

19. The method of claim 18, wherein the release of heparin from the nanoparticle is controlled by the application of radiation at a wavelength in the range of about 700 nm to about 1000 nm.

20. The method of claim 19, wherein the heparin is locally released from the nanoparticle by heating the nanoparticle so that it alters the ultrastructure of the polymer.

21. The method of claim 14, wherein the nanospun matrix further comprises an image enhancing agent.

22. The method of claim 21, wherein the image enhancing agent is gadolinium.

23. The method of claim 1, wherein the endothelial cells are isolated from a human saphenous vein.

24. The method of claim 1, wherein the endothelial cells are derived from progenitor cells isolated from peripheral blood or bone marrow.

25. A preconditioned tissue engineered heart valve, comprising:

a biocompatible matrix having a heart valve shape seeded with a population of endothelial cells differentiated from progenitor cells, wherein the endothelial cells attach to the matrix to form an endothelial cell layer;

a population of smooth muscle cells differentiated from progenitor cells seeded onto the endothelial cell layer, wherein the smooth muscle cells form a smooth muscle cell layer that attaches to the endothelial cell layer; and

wherein the seeded cells have been preconditioned in a preconditioning chamber that exposes the seeded cells to a biological fluid that is passed through the seeded matrix at a flow-rate and pulse-rate equivalent to normal blood flow through the heart.

26. The method of claim 25, wherein biocompatible matrix is selected from the group consisting of a decellularized matrix, an electrospun matrix, and a synthetic polymer matrix.

27. The method of claim 25, wherein biocompatible matrix is a decellularized heart valve.