Decellularized neuromuscular scaffold를 이용한 근육결손부 재건 연구 : A study on recovery of muscle defect using decellularized neuromuscular scaffold

Abstract

Introduction It is known that there is more effective results to transplant the neuromuscular tissue than nerve alone for regeneration of damaged neuromuscular function. Therefore it would be necessary to acellular neuromuscular graft for recovery of effective myofunction. However the study for decellularization of skeletal muscle with motor nerve is rare. This study aims to establish a basis for attempting homologous transplantation from acellular neuromuscular scaffold of rats. Methods 8-weeks male Sprague rats (SD rat, SPF) were operated to hervest some part of extensor digitorum longus (EDL) and tibialis anterior (TA) muscle with peroneal nerve. Acellular neuromuscular scaffolds were designed by three decellularization protocols which using trypsin and triton X-100 (protocol I), using Sulfobetaine(SB)-10, SB-16 and and triton X-100 (protocol II ) and using SB-10, trypsin and triton X-100 (protocol III). The degree of decellularization of the produced scaffold is evaluated and transplanted into the neuromuscular defect which was made on the homologous rat. After clinical evaluation of peroneal nerve function in rats at 2 weeks after transplantation, the rats were sacrificed for histological analysis. Result In this study, the protocol III using SB-10, Trypsin, and Triton X-100, which are presented in this study, seems to be decellularized at an appropriate level which removed DNA of neuromuscular tissue and showed proper network structure. In addition, it showed no immunologic response in allografts and good biocompatibility with peripheral muscles. The acellular neuromuscular scaffold produced by the protocol III was found to be successful result in histologic and clinical evaluation at 2 weeks after allograft. In order to use more clinical applications, further study for more specimens, population and long-term follow-up should be necessary to confirm the nerve reinnervation after transplantation of acellular neuromuscular scaffold with electromyography, histologic-chemical and histologic-enzymal staining.