Electroporation of tissue products
View Patent ↗The present disclosure provides methods for reducing bioburden on a tissue product, as well as the tissue products produced according to the disclosed methods. In particular, the disclosure relates to methods of electroporating tissue in the presence of one or more bactericides in order to reduce bioburden. The methods allow for reduced exposure to electrical energy and/or bactericide while reducing bioburden.
1. A method of reducing the bioburden in a tissue product, comprising
providing a human or animal tissue;
contacting the tissue with one or more bactericides; and
exposing the tissue to one or more electrical pulses.
2. The method of claim 1 , wherein the method produces a substantial reduction in bioburden on the tissue.
3. The method of claim 2 , wherein the bioburden on the tissue is reduced by at least about 50%.
4. The method of claim 1 , wherein the duration of the electrical pulses, voltage of the electrical pulses, and number of electrical pulses are controlled such that the extracellular matrix of the tissue product is not damaged during exposure to the electrical pulses.
5. The method of claim 1 , wherein the duration of exposure to and concentration of the one or more bactericides are controlled such that the extracellular matrix of the tissue product is not damaged during exposure to the one or more bactericides.
6. The method of claim 1 , wherein the duration of the electrical pulses, voltage of the electrical pulses, and number of electrical pulses are controlled such that the temperature of the tissue product does not rise above about 42 degrees Celsius.
7. The method of claim 1 , wherein the duration of the electrical pulses, voltage of the electrical pulses, and number of electrical pulses are controlled such that the pH of the tissue product does not rise above about pH 8.
8. The method of claim 1 , wherein the one or more electrical pulses have a voltage of about 1 volt (V) to about 10 kilovolts (kV).
9. The method of claim 1 , wherein the one or more electrical pulses have a voltage of about 40V or 80V.
10. The method of claim 1 , further comprising administering the one or more electrical pulses for about 1 millisecond (ms) to about 1 second.
11. The method of claim 1 , wherein the one or more electrical pulses have a duration of about 5 ms, about 10 ms, or about 15 ms.
12. The method of claim 1 , comprising administering about 1 to 100,000 electrical pulses.
13. The method of claim 1 , comprising administering about 1 to 500 electrical pulses.
14. The method of claim 1 , further comprising allowing the tissue to cool and then exposing the tissue to one or more additional electrical pulses.
15. The method of claim 14 , further comprising repeating the exposure to one or more additional electrical pulses 2-5 times.
16. The method of claim 1 , wherein the bactericide comprises one or more microbial growth inhibitors, cytotoxic agents, oxidants, and/or antibiotics.
17. The method of claim 1 , wherein the bactericide comprises one or more of a peroxide, oxidizer, antimicrobial metal, quaternary ammonium compound, or charged bactericidal compound.
18. The method of claim 1 , wherein the bactericide comprises one or more of peracetic acid (PAA), ozone, hypochlorite, silver, zinc, copper, benzalkonium chloride, cetylpyridinium chloride, benzethonium chloride, cetyltrimethyl ammonium bromide, or chitosan.
19. The method of claim 1 , wherein the bactericide comprises PAA.
20. The method of claim 19 , wherein the PAA is at a concentration of about 0.01%-2% weight/volume.
21. The method of claim 19 , wherein the PAA is at a concentration of about 0.2% weight/volume.
22. The method of claim 1 , wherein the bactericide is applied to the tissue at a concentration sufficient to substantially reduce bioburden on the tissue.
23. The method of claim 1 , further comprising irradiating the tissue using e-beam radiation.
24. The method of claim 1 , further comprising decellularizing the tissue.
25. The method of claim 24 , wherein the tissue is decellularized by contact with one or more detergents and by exposure to the one or more electrical pulses.