Preparation Of The Acellular Matrix In The Tissue Engineering And Regenerative Medicine And Study Of Biocompatibility

Tissue engineering and regenerative medicine are the advanced technologies for treating the failure of tissues and organs. They are better than the traditional therapy.The basis of these technologies is cellular matrix. It must have biocompatibility, surface compatibility, structure compatibility. biodegradation,and it also can be sterilized.Because people can control the synthetic matrix appearance, dimension, intensity degradation rate and three dimensional,they used to use it.But it also brings biocompatibility and infection .Following the development of the study in extracellular matrix (ECM), people find that the ECM is composed of glucoprotein and glycosaminoglycans, and it's components induct the cell differentiation in tissues and organs, in turn, it can modulate the function of these. People find that ECM is the best matrix in tissue engineering and regenerative medicine.but people can't produce the matrix which is same as ECM.Now people find that the components and structures of ECM in homologuous tissues and organs are identifiable,even in the heterogenous ones.And this kind of ECM are used extensively in the fields for example the skin,urology orthopaedics etc.This kind of ECM can be got fran decellizing the tissues and organs, it also called acellular matrix. But the decellation techulogy is incompleted now. It can't get rid of all cells in tissues and organs some times, and decellation maybe do harm to matrix.The preparation of acellular matrix includes decella-tion, technology which is used to remore cell debris and enhance theintensity of matrix. Trypsin, Dispase or hypertonic saline is used in decelliztion. Triton X-100 and Sodiumdodecylsulphate (SDS) are effective detergent in removing cell debris, but it is unknom that which of these is more effective. The study of enhancing the intensity of matrix is the new field. In one word, the methods for producing acellulace matrix have not been standardized.In this study, we want to select the best way to produce acellular matrix. We also use immunohistochemical method for detecting the magor heterogenetic antigen a -galactosyl residues( a -Gal) in acellular matrix.We treat the porcine skin by 0.25 percent trypsin, 0.125% trypsin, 2.5 U/ml Dispase,hypertonic saline or hypertonic saline-trypsin/Dispase.We find that after the skin has been incubated in 0.125 percent trypsin for 24h at 4癈,The cells in the skin are all disintegrated .there are no significant differentiation between the acellular matrix treated by 0.125, 0.25 perlent trypsin,2.5 U/ml Dispase and hypertonic saline-trypsin/Dispase.But the cell can't be removed by using the hypertonic saline-SDS .Subsequently the acellular matrix is incubated in 0.5 percent SDS for 3h at room temperature with contiuous shaking. The cell debris are removed completely by this way, but the intensity of matrix will be decreased significantly. The cell debris can't be removed by using 0.5 percent Triton X-100 or the solution which is mixed by 0.5 percent Triton X-100 and 0.5 percent SDS at the ratio of 1:1.25, 1:1, 1:0.75, 1:0.5, 1:0.25, 1:0.125.Glutaraldehyde can increased the intensity of acellular matrix significantly. If the skin has been put into 4 percent glutaraldehydefor three minutes or more. We are no longer able to get rid of the cells in it.The a -Gal in porcine skin as disapered in acellular matrix treated by trypsin,Dispase and SDS.The acellular matrix treated by SDS or glutaraldehyde should be cleaned by normal saline, or there will be cytotoxicity in it. The results of cell culture and implantation experiment demonstrate the high degree of cytocompatibibity and biocompatibility of this kind of acelluler matrix.