Cell-free tissue replacement for tissue engineering
View Patent ↗The present invention is a natural, cell-free tissue replacement that does not require difficult or extensive preparation made by washing tissue replacement in a solution including one or more sulfobetaines and an anionic surface-active detergent and washing the tissue replacement in serial solutions of the buffered salt to remove excess detergent. The natural, cell-free tissue replacement may be a nerve graft that supports axonal regeneration, guides the axons toward the distal nerve end and/or is immunologically tolerated. Other forms of the invention are a composition and kit prepared by the method of making a native, cell-free tissue replacement. The present invention may be modified for use in diagnostic, therapeutic, and prophylactic applications.
1. A method for preparing a native, acellular nerve tissue replacement comprising the steps of:
obtaining a nerve tissue;
soaking the nerve tissue for at least six hours in a solution comprising one or more sulfobetaines;
treating the nerve tissue in a mixture of one or more sulfobetaines and Triton X-200; and
washing the nerve tissue in one or more solutions of a buffered salt to remove excess detergent to form the native, acellular nerve tissue replacement with significantly reduced immunologic response relative to a nerve tissue graft made acellular by a freeze/thaw process or a decellularization process utilizing Triton X-100.
2. The method of claim 1 , further comprising the step of storing the native, acellular nerve tissue replacement in a buffered salt solution until needed.
3. The method of claim 1 , wherein the sulfobetaines have hydrophilic tails of 10 to 16 carbons.
4. The method of claim 1 , further comprising the step of:
adhering one or more bioactive agents to the tissue replacement.
5. The method of claim 4 , wherein the one or more bioactive compounds comprises a drug.
6. The method of claim 1 , wherein the native, acellular nerve tissue replacement further comprises a structure selected from the group consisting of a tube, sheet, film, scaffold, and tissue transplant for delivery into the body.
7. The method of claim 1 , wherein the sulfobetaine comprises SB-16.
8. The method of claim 1 , wherein the step of washing the nerve tissue comprises one or more washes in a buffered salt solution comprising 100 mM sodium and 50 mM phosphate for at least 15 minutes each.
9. The method of claim 1 , wherein the nerve tissue is harvested from a mammalian cadaver.
10. The method of claim 9 , wherein the nerve tissue is cleaned of fat and blood after harvesting and rinsed two or more times in deionized distilled water.
11. A native, acellular nerve tissue replacement with significantly reduced immunologic response made by the method of claim 1 .
12. A kit for tissue replacement comprising the native, acellular nerve tissue replacement with significantly reduced immunologic response of claim 11 .
13. The kit of claim 12 , wherein the native, acellular nerve tissue replacement further comprises a tube, a sheet, a film, a scaffold, or a nerve tissue transplant.
14. The kit of claim 13 , wherein the native, acellular nerve tissue replacement further comprises a polymer, a bioactive compound or combinations thereof.
15. The kit of claim 13 , further comprising a sheet of instructions for use of the native, acellular nerve tissue replacement.
16. A method for preparing a native, acellular nerve tissue replacement comprising the steps of:
obtaining a nerve tissue;
soaking the nerve tissue for at least six hours in a solution comprising one or more sulfobetaines;
treating the nerve tissue in a mixture of one or more sulfobetaines and Triton X-200; and
washing the nerve tissue in one or more solutions of a buffered salt to remove excess detergent to form the native, acellular nerve tissue replacement, wherein the basal laminae and endoneurium layer substantially retain the native extracellular matrix structure.
17. The method of claim 16 , wherein the native, acellular nerve tissue replacement, when implanted, elicits a T-cell mediated immune response that is less than an immune response triggered by an allogeneic implant.
18. The method of claim 16 , wherein the native, acellular nerve tissue replacement allows for higher axon density when implanted relative to a nerve tissue graft made acellular by a freeze/thaw process or a decellularization process utilizing Triton X-100.