Method for decellularising foreign material for the production of bioprostheses

Abstract

The invention relates to a method for decellularising allogenic and xenogenic foreign material for the subsequent production of bioprostheses, coated with endogenous body cells, whereby the foreign material is firstly treated in a solution of bile acid and then alcohol, each with an intermediate or subsequent rinsing step, in combination with a mechanical action on the tissue and the cells by the force generated from a flowing treatment medium, at least in the final rinsing step. After said treatment the tissue is completely rid of foreign cell material and viruses and represents an excellent starting material for the coating with cells of the bioprosthesis recipient.

1 . A method for decellularizing allogenic and xenogenic foreign material using biodetergents to produce bioprostheses coated with endogenic cells from the recipient of the prosthesis, characterized in that the foreign material is initially treated with bile acid to kill the cells and to coat the cells as well as to separate the bond with the tissue matrix, then treated with alcohol to kill more cells, each of these steps being followed by a rinsing step and at least the last rinsing step being executed in a media flow so that the fluid forces act mechanically on the tissue matrix and the cells.  
     
     
         2 . The method according to  claim 1  wherein the media flow is pulsating.  
     
     
         3 . The method according to  claim 1 , characterized in that a cell-encompassing deoxycholic acid is used to detach the cells with bile acid.  
     
     
         4 . The method according to  claim 3  wherein 1% to 2% deoxycholic acid is used.  
     
     
         5 . The method according to  claim 1  wherein 30% to 60% alcohol is used to kill the cells.  
     
     
         6 . The method according to  claim 5  wherein 40% alcohol is used.  
     
     
         7 . The method according to  claim 1  wherein a phosphate buffer solution is used as rinsing medium.  
     
     
         8 . The method according to  claim 7  wherein decreasing concentrations of the rinsing medium are used for rinsing with non-flowing medium.  
     
     
         9 . The method according to claims  1  through  7  characterized in that the deoxycholic acid treatment in non-flowing medium takes up to 24 hours, alcohol treatment takes about 20 minutes, and rinsing takes about 60 minutes.  
     
     
         10 . The method according to claims  1  through  7  characterized in that the treatment of the foreign material in deoxycholic acid and/or alcohol and/or the intermediate rinsing cycle are carried out in a pulsating medium flow that comes close to the natural flow conditions in the respective organ.  
     
     
         11 . The method according to claims  1  and  10  wherein the speed of the flowing media can be varied.  
     
     
         12 . The method according to claims  1 ,  10  and  11  characterized in that the flowing rinsing medium is replaced continuously or discontinuously.  
     
     
         13 . The method according to  claim 11 , characterized in that the concentration of the detergent flow is gradually reduced.  
     
     
         14 . The method according to any one of claims  1  and  10  through  13 , characterized in that the foreign material to be treated is held in a preloaded state in the respective media flow.  
     
     
         15 . An apparatus for treating foreign material in a media flow according to claims  1  and  10  through  14 , characterized by a ring line ( 1 ) with a pump ( 3 ) connected to it to generate the pulsating media flow and a decellularization chamber ( 2 ) detachably integrated into the ring line ( 1 ) in which the allogenic or xenogenic foreign material ( 8 ) to be treated can be fixed lengthwise in a preloaded state in the direction of the treatment media flow using adapters ( 10 ,  11 ) that are detachably mounted to the decellularization chamber ( 2 ) and comprise a fixing section ( 14 ,  15 ) and fittings ( 12 ,  13 ) for connecting the ring line ( 1 ).