Preparation of human decellularized peripheral nerve allograft using amphoteric detergent and nuclease  

作　　者：Joo-Yul Bae Suk Young Park Young Ho Shin Shin Woo Choi Jae Kwang Kim 

机构地区：[1]Department of Orthopedic Surgery,University of Ulsan College of Medicine,Gangneung Asan Hospital,Gangneung-si,Korea [2]Department of Orthopedic Surgery,University of Ulsan College of Medicine,Asan Medical Center,Seoul,Korea

出　　处：《Neural Regeneration Research》2021年第9期1890-1896,共7页中国神经再生研究（英文版）

基　　金：supported by the Korea Health Industry Development Institute,No.HI17C1221(to JKK);National Research Foundation of Korea,Nos.NRF-2017R1A2B4003692,NRF-2020R1A2C1006656(both to JKK);the Asan Institute for Life Sciences,Asan Medical Center,Nos.2018-766,2019IP0766(both to JKK)。

摘　　要：Animal studies have shown that amphoteric detergent and nuclease(DNase I and ribonuclease A) is the most reliable decellularization method of the peripheral nerve. However, the optimal combination of chemical reagents for decellularization of human nerve allograft needs further investigation. To find the optimal protocol to remove the immunogenic cellular components of the nerve tissue and preserve the basal lamina and extracellular matrix and whether the optimal protocol can be applied to larger-diameter human peripheral nerves, in this study, we decellularized the median and sural nerves from the cadavers with two different methods: nonionic and anionic detergents(Triton X-100 and sodium deoxycholate) and amphoteric detergent and nuclease(3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate(CHAPS), deoxyribonuclease I, and ribonuclease A). All cellular components were successfully removed from the median and sural nerves by amphoteric detergent and nuclease. Not all cellular components were removed from the median nerve by nonionic and anionic detergent. Both median and sural nerves treated with amphoteric detergent and nuclease maintained a completely intact extracellular matrix. Treatment with nonionic and anionic detergent decreased collagen content in both median and sural nerves, while the amphoteric detergent and nuclease treatment did not reduce collagen content. In addition, a contact cytotoxicity assay revealed that the nerves decellularized by amphoteric detergent and nuclease was biocompatible. Strength failure testing demonstrated that the biomechanical properties of nerves decellularized with amphoteric detergent and nuclease were comparable to those of fresh controls. Decellularization with amphoteric detergent and nuclease better remove cellular components and better preserve extracellular matrix than decellularization with nonionic and anionic detergents, even in large-diameter human peripheral nerves. In Korea, cadaveric studies are not yet legally subject to Institutional Review Board rev

关 键 词：median nerve sural nerve NUCLEASE DETERGENT human decellularized nerve graft 

分 类 号：R741.05[医药卫生—神经病学与精神病学]