Preliminary research on preparation of porcine bladder acellular matrix graft for tissue engineering applications

Abstract

To investigate the preparation method of porcine bladder acellular matrix graft (BAMG) and evaluate the feasibility of using BAMG as biomaterial scaffold to construct tissue engineering bladder. Bladders were harvested from adult pigs at a slaughterhouse and were decellularized by the method of enzyme digestion or eradicator washing. The BAMG was then examined by optic microscope and electron microscope to confirm no cell elements remained. The structure of the BAMG was always observed through the Scanning Microscope. The skeletal muscle stem cells of New Zealand white rabbits were implanted in 96-hole-plank and cultured by BAMG extract liquid or normal culture medium. The cytotoxicity of BAMG was tested through MTT assay. New Zealand white rabbits were underwent bladder triangle area spared cystectomy and reconstructed by the heterogeneity BAMG. The animal bladders were obtained after six weeks and the tissue regeneration was examined. Good outcome were obtained with either of the two methods. There were no cell elements remained under the examination of optical microscope and electron microscope. The interstice could observed on BAMG through the Scanning Microscope. The relative growth rate (RGR) was 81%, 88%, 93% and 96% separately on the 2nd, the 4th, the 6th and the 8th day (t = 3.7419, P < 0.025). The cytotoxicity score was I, which indicated that BAMG had a good cytocompatibility. All rabbits were alive except one which died from diarrhea. The reconstructed bladder resumed the construction and function of the bladder. The transitional cells and muscle cells growth could be seen in the scaffold of the repaired bladder after six weeks. The porcine BAMG has good biocompatibility and can be used as ideal biomaterial scaffold of bladder tissue engineering.