Electroporation of tissue products

Abstract

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 - 38 . (canceled) 
     
     
         39 . A method of reducing a bioburden in a tissue product, comprising:
 providing a human or animal tissue;   exposing the human or animal tissue to one or more electrical pulses; and   controlling at least one of a pulse voltage, a pulse duration, or a number of pulses, of the electrical pulses to regulate a pore size in the human or animal tissue.   
     
     
         40 . The method of  claim 39 , comprising controlling at least one of the pulse voltage, the pulse duration, or the number of pulses, of the electrical pulses to regulate an amount of time that pores in the human or animal tissue remain open. 
     
     
         41 . The method of  claim 39 , comprising contacting the human or animal tissue with one or more bactericides. 
     
     
         42 . The method of  claim 39 , comprising applying the one or more electrical pulses for a long duration, at a high voltage or in multiple pulses in rapid succession to create temporary or permanent pores in lipid membranes of the human or animal tissue. 
     
     
         43 . The method of  claim 41 , wherein exposing the human or animal tissue to the one or more electrical pulses drives the one or more bactericides into the human or animal tissue. 
     
     
         44 . The method of  claim 41 , wherein exposing the human or animal tissue to the one or more electrical pulses evenly administers the one or more bactericides across a thickness of the human or animal tissue. 
     
     
         45 . The method of  claim 41 , wherein the method produces a substantial reduction in the bioburden on the human or animal tissue. 
     
     
         46 . The method of  claim 45 , wherein the bioburden on the human or animal tissue is reduced by at least about 50%. 
     
     
         47 . The method of  claim 39 , wherein the pulse duration, the pulse voltage, or the number of pulses is controlled such that an extracellular matrix of the tissue product is not damaged during exposure to the one or more electrical pulses. 
     
     
         48 . The method of  claim 41 , wherein a duration of exposure to and concentration of the one or more bactericides are controlled such that an extracellular matrix of the tissue product is not damaged during exposure to the one or more bactericides. 
     
     
         49 . The method of  claim 39 , wherein the pulse duration, the pulse voltage, or the number of pulses is controlled such that a temperature of the tissue product does not rise above about 42 degrees Celsius. 
     
     
         50 . The method of  claim 39 , wherein the pulse duration, the pulse voltage, or the number of pulses is controlled such that a pH of the tissue product does not rise above about pH 8. 
     
     
         51 . The method of  claim 39 , wherein the pulse voltage of the one or more electrical pulses is about 1 volt (V) to about 10 kilovolts (kV). 
     
     
         52 . The method of  claim 39 , wherein the pulse voltage of the one or more electrical pulses is about 40V or 80V. 
     
     
         53 . The method of  claim 39 , comprising administering the one or more electrical pulses for the pulse duration of about 1 millisecond (ms) to about 1 second. 
     
     
         54 . The method of  claim 39 , wherein the pulse duration of the one or more electrical pulses is about 5 ms, about 10 ms, or about 15 ms. 
     
     
         55 . The method of  claim 39 , wherein the number of pulses administered is about 1 to 100,000 electrical pulses. 
     
     
         56 . The method of  claim 39 , wherein the number of pulses administered is about 1 to 500 electrical pulses. 
     
     
         57 . The method of  claim 39 , comprising allowing the human or animal tissue to cool and then exposing the human or animal tissue to one or more additional electrical pulses. 
     
     
         58 . The method of  claim 57 , comprising repeating the exposure to one or more additional electrical pulses 2-5 times. 
     
     
         59 . The method of  claim 41 , wherein the bactericide comprises one or more microbial growth inhibitors, cytotoxic agents, oxidants, and/or antibiotics. 
     
     
         60 . The method of  claim 41 , wherein the bactericide comprises one or more of a peroxide, oxidizer, antimicrobial metal, quaternary ammonium compound, or charged bactericidal compound. 
     
     
         61 . The method of  claim 41 , 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. 
     
     
         62 . The method of  claim 41 , wherein the bactericide comprises PAA. 
     
     
         63 . The method of  claim 62 , wherein the PAA is at a concentration of about 0.01%-2% weight/volume. 
     
     
         64 . The method of  claim 62 , wherein the PAA is at a concentration of about 0.2% weight/volume. 
     
     
         65 . The method of  claim 41 , wherein the bactericide is applied to the human or animal tissue at a concentration sufficient to substantially reduce bioburden on the human or animal tissue. 
     
     
         66 . The method of  claim 39 , comprising irradiating the human or animal tissue using e-beam radiation. 
     
     
         67 . The method of  claim 39 , further comprising decellularizing the human or animal tissue. 
     
     
         68 . The method of  claim 67 , wherein the human or animal tissue is decellularized by contact with one or more detergents and by exposure to the one or more electrical pulses.