Chemical treatment for removing cellular and nuclear material from naturally occurring extracellular matrix-based biomaterials

Abstract

A method is provided for sequential decellularization of an isolated tissue using solubilizing solutions comprising at least one oxidizing agent that removes all cellular and nuclear materials from the tissue while substantially maintaining the biological and, mechanical properties, and the biochemical properties of the resulting extracellular matrix.

1 . A biocompatible tissue implant, comprising a network of extracellular matrix (ECM) derived from submucosa treated with a first solubilizing fluid comprising at least one oxidizing agent at a concentration effective to extract cellular and nuclear material while substantially maintaining the biochemical composition and the mechanical and biological properties of the tissue, and a second solubilizing fluid comprising at least one oxidizing agent, to further remove cellular and nuclear material while substantially maintaining the biochemical composition and the mechanical and biological properties of the small intestinal submucosa, and a washing fluid to remove cellular debris, wherein the matrix consists of a framework of collagen substantially free of cellular material and viable and non-viable nucleic acids, and wherein the matrix is effective to permit the growth of cells. 
   
   
       2 . The implant of  claim 1 , wherein the ECM is approximately 20% substantially free of viable and non-viable nucleic acids. 
   
   
       3 . The implant of  claim 1 , wherein the ECM is approximately 30% substantially free of viable and non-viable nucleic acids. 
   
   
       4 . The implant of  claim 1 , wherein the ECM is approximately 40% substantially free of viable and non-viable nucleic acids. 
   
   
       5 . The implant of  claim 1 , wherein the ECM is approximately 50% substantially free of viable and non-viable nucleic acids. 
   
   
       6 . The implant of  claim 1 , wherein the ECM is approximately 60% substantially free of viable and non-viable nucleic acids. 
   
   
       7 . The implant of  claim 1 , wherein the ECM is approximately 70% substantially free of viable and non-viable nucleic acids. 
   
   
       8 . The implant of  claim 1 , wherein the ECM is approximately 80% substantially free of viable and non-viable nucleic acids. 
   
   
       9 . The implant of  claim 1 , wherein the ECM is approximately 90% substantially free of viable and non-viable nucleic acids. 
   
   
       10 . The implant of  claim 1 , wherein the ECM is approximately 100% substantially free of viable and non-viable nucleic acids. 
   
   
       11 . The implant of  claim 1 , wherein the viable and non-viable nucleic acids are measured using a PicoGreen® assay. 
   
   
       12 . The implant of  claim 11 , wherein the viable and non-viable nucleic acid content is about 50 micrograms/mg to about 0.5 microgram/mg measured using the PicoGreen® assay. 
   
   
       13 . The implant of  claim 11 , wherein the viable and non-viable nucleic acid content is about 40 micrograms/mg to about 0.4 microgram/mg measured using the PicoGreen® assay. 
   
   
       14 . The implant of  claim 11 , wherein the viable and non-viable nucleic acid content is about 30 micrograms/mg to about 0.3 microgram/mg measured using the PicoGreen® assay. 
   
   
       15 . The implant of  claim 11 , wherein the viable and non-viable nucleic acid content is about 20 micrograms/mg to about 0.2 microgram/mg measured using the PicoGreen® assay. 
   
   
       16 . The implant of  claim 11 , wherein the viable and non-viable nucleic acid content is about 10 micrograms/mg to about 0.1 microgram/mg measured using the PicoGreen® assay. 
   
   
       17 . The implant of  claim 11 , wherein the viable and non-viable nucleic acid content is about 35 micrograms/mg to about 3.5 microgram/mg measured using the PicoGreen® assay.