Fabrication of living blood vessels and glandular tissues

• US 4,546,500 A
• Filed: 02/26/1982
• Issued: 10/15/1985
• Est. Priority Date: 05/08/1981
• Status: Expired due to Term

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Bell, Eugene
• Primary Examiner(s): Apley, Richard J.
• Assistant Examiner(s): ISABELLA, DAVID J
• Application Number:
  US06/352,585
• Time in Patent Office: 1,327 Days
• Field of Search: 3/1, 3/1.4, 435/240-241, 435/1, 128/DIG. 8
• US Class Current: 435/1.1
• CPC Class Codes: A61F 2/022 Artificial gland structures using bioreactors hemodialysis A61M1/16; hemofiltration A61M1/34; other extracorporeal blood treatment A61M1/36
• CPC Class Codes: A61F 2/06 Blood vessels
• CPC Class Codes: A61F 2/062 Apparatus for the production of blood vessels made from natural tissue or with layers of living cells prostheses made from natural tissue or living cells A61L27/507
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/36 containing ingredients of undetermined constitution or reaction products thereof , e.g. transplant tissue, natural bone, extracellular matrix isolated natural compounds, e.g. collagen A61L27/24
• 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 623/901 Method of manufacturing prosthetic device
• CPC Class Codes: Y10S 623/921 Blood vessel

Abstract

A method and apparatus for producing a vessel-equivalent prosthesis is described. A contractile agent such as fibroblast cells, smooth muscle cells or platelets is incorporated into a collagen lattice and contracts the lattice axially around an inner core. After the structure has set, additional layers may be formed in an ordered manner depending on the intended function of the prosthesis. Alternatively, all the layers may be formed concurrently. A plastic mesh sleeve is sandwiched between layers or embedded within the smooth muscle cell layer to reinforce the structure and provide sufficient elasticity to withstand intravascular pressure.

Claims

1. In a method of producing, in vitro, a living multi-layered tubular structure, comprising:

a. producing a first tubular layer by;

(1) forming a first aqueous mixture of collagen fibrils, nutrient medium and a first cellular contractile agent capable of interacting with collagen fibrils;

(2) introducing said first mixture into an annular casting chamber;

(3) maintaining the annular casting chamber containing said first mixture under conditions sufficient to allow a gel to form therein and to allow radial contraction of the gel with expression of aqueous liquid therefrom resulting from interaction of the cellular contractile agent with collagen fibrils to thereby form a contracted hydrated collagen lattice suitable as one layer of a multi-layered tubular structure; and

(4) removing aqueous liquid expressed in the formation of said first tubular layer from the annular casting chamber;

b. producing a second tubular layer outwardly of said first tubular layer by;

(1) forming a second aqueous mixture of collagen fibrils, nutrient medium and a second cellular contractile agent capable of interacting with collagen fibrils;

(2) introducing said second mixture into the annular casting chamber;

(3) maintaining the annular casting chamber containing said second mixture under conditions sufficient to allow a gel to form therein and to allow radial contraction of the gel with expression of aqueous liquid therefrom resulting from interaction of the cellular contractile agent with collagen fibrils to thereby form a contracted hydrated collagen lattice suitable as another layer of a multi-layered tubular structure; and

c. removing said multi-layered tubular structure from the annular casting chamber;

The improvement comprising adding a reinforcing sleeve of inert material to said living multi-layered tubular structure.

2. The improvement of claim 1 wherein said reinforcing sleeve of inert material comprises a plastic mesh sleeve.

3. The improvement of claim 2 wherein said plastic mesh sleeve is located between the first and second tubular layers of the living multi-layered tubular structure.

4. The improvement of claim 3 wherein the plastic mesh sleeve is attached by first slipping it on a tube and subsequently sliding the tube over the first tubular structure and removing the tube leaving the sleeve positioned over the first tubular structure.

5. The improvement of claim 3 wherein said plastic mesh sleeve is pretreated to render it more electronegative.

6. The improvement of claim 5 wherein said plastic mesh screen is pretreated by subjecting it to plasma.

7. A living multi-layered tubular structure produced by the improvement of claim 1.

8. A living multi-layered tubular structure produced by the improvement of claim 3.

10. The improvement of claim 8 wherein said plastic mesh comprises a sleeve positioned between the layers of smooth muscle cells and fibroblast cells.

11. The improvement of claim 10 wherein said plastic mesh screen comprises polyethylene terephthalate.

12. The improvement of claim 11 wherein said plastic mesh screen is pretreated to make it more electronegative.

13. The improvement of claim 12 wherein said pretreatment is done by exposing the plastic mesh screen to plasma.

9. In a method of producing a living prosthesis, in vitro, comprising the steps of:

a. fabricating a cylindrical smooth muscle cell layer as follows;

(aa) separately preparing (i) an aqueous acidic mixture comprising nutrient medium and collagen fibrils and (ii) a mixture of smooth muscle cells suspended in nutrient medium;

(ab) raising the pH of mixture (i) and quickly combining mixture (i) and mixture (ii) and pouring the combined mixture into a casting chamber having an inner core member and an outer cylindrical wall structure to form a lattice;

(ac) incubating the lattice for a period sufficient to enable collagen fibrils to be compacted by the cells so that aqueous liquid is expressed out of the lattice as the lattice contracts radially about the core;

(ad) removing aqueous liquid expressed in step (ac);

(ae) repeating steps (aa)-(ad) if additional layers of smooth muscle cells are desired;

b. fabricating a layer containing fibroblast cells on said cylindrical smooth muscle layer as follows;

(ba) separately preparing (i) an aqueous acidic mixture comprising nutrient medium and collagen fibrils and (ii) a mixture of fibroblast cells suspended in nutrient medium;

(bb) raising the pH of mixture (i) and quickly combining mixture (i) and mixture (ii) and pouring the combined mixture into a casting chamber having as an inner core member of cylindrical smooth muscle cell layer and an outer cylindrical wall structure to form a lattice;

(bc) incubating the lattice formed in step (bb) in accordance with step (ac);

(bd) removing aqueous liquid expressed in step (bc);

c. lining the inner wall of the cylindrical smooth muscle cell layer with living cells;

The improvement of including a plastic mesh within said living prosthesis to thereby reinforce said living prosthesis and to provide it with an increased degree of elasticity.

14. A living multi-layered tubular structure produced by the improvment of claim 9.

15. In a tubular prosthesis formed of multiple layers of hydrated collagen lattices contracted with living cells:

The improvement comprising including a plastic mesh sleeve embedded in the layers of said prosthesis to provide reinforcement and elasticity thereto.

16. The improvement of claim 15 wherein said plastic mesh sleeve is formed from polyethylene teraphthalate.