低免疫原性脱细胞神经基质的制备及其理化性能

doi:10.3969/j.issn.2095-9400.2020.06.001

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Abstract Objective: To prepare low immunogenic xenogeneic decellularized nerve matrix and evaluate its biological properties. Methods: To prepare low immunogenic xenogeneic decellularized nerve matrix by decellularizing the α-1, 3-galactoside transferase gene-knockout (GTKO) pig sciatic nerve with a self-designed combination protocol of hypotonic solution, trypsin, detergent, and nuclease. The decellularization was evaluated by HE, DAPI staining, and the amount of residual DNA. Immunohistochemical staining of type-I collagen, laminin, fibronectin, and Sirius scar staining were used to assess the retention of extracellular matrix components of acellular nerve. A-gal immunofluorescence was used to detect the residual of the heterologous antigen, a-gal. Finally, the three-dimensional (3D) structure and porosity of acellular nerves were analyzed by scanning electron microscopy (SEM). Results: DNA content in the decellularized nerve tissue was 95% lower than that in the natural nerve tissue, and there were few cellular components in the endoneurium, perineurium, and epineurium of acellular nerve. Sirius scar staining and immunohistochemistry showed that the decellularized nerve tissue retained collagen fibers and major extracellular matrix proteins. SEM results showed that the acellular nerve retained its three-dimensional (3D) structure. Cytotoxicity tests showed that the acellular nerve matrix was not cytotoxic. Conclusion: The self-designed decellularization protocol of this study can completely remove the cellular components and heterologous antigens in the nervous tissue, and retain the neural tissue structure more completely. With high similarity to the natural nerves, it has great potential for application as nerve grafts in the future.

Key words： decellularization low immunogenicity extracellular matrix nerve grafts gene knockout

Received: 20 September 2019

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	WANG Jianying
	LI Fanghong
	LI Huaqiong
	WANG Ronghua
	WANG Chenfei
	ZHAO Allan Zijian

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https://xb.wmu.edu.cn/EN/10.3969/j.issn.2095-9400.2020.06.001 OR https://xb.wmu.edu.cn/EN/Y2020/V50/I6/431

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