Open AccessVarious changes in cryopreserved acellular nerve allografts at −80°C
Author(s) -
Yanyan Huang,
Xiaoting Xu,
Xijun Huang,
Jianghui Liu,
Jian Qi,
Shuang Zhu,
Zhaowei Zhu,
Bo He,
Qian Zhu,
Yangbin Xu,
Liqiang Gu,
Xiaolin Liu
Publication year - 2018
Publication title -
neural regeneration research/neural regeneration research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.93
H-Index - 38
eISSN - 1876-7958
pISSN - 1673-5374
DOI - 10.4103/1673-5374.237138
Subject(s) - cryopreservation , decellularization , transplantation , cryoprotectant , neurofilament , sciatic nerve , pathology , anatomy , h&e stain , dimethyl sulfoxide , gastrocnemius muscle , andrology , chemistry , staining , medicine , immunohistochemistry , tissue engineering , biology , biomedical engineering , surgery , microbiology and biotechnology , embryo , skeletal muscle , organic chemistry
The experimental design evaluated histological, mechanical, and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle, multi-directional manner to provide evidence for long-term preservation. Acellular nerve allografts from human and rats were cryopreserved in a cryoprotectant (10% fetal bovine serum, 10% dimethyl sulfoxide, and 5% sucrose in RPMI1640 medium) at -80°C for 1 year, followed by thawing at 40°C or 37°C for 8 minutes. The breaking force of acellular nerve allografts was measured using a tensile test. Cell survival was determined using L-929 cell suspensions. Acellular nerve allografts were transplanted into a rat model with loss of a 15-mm segment of the left sciatic nerve. Immunohistochemistry staining was used to measure neurofilament 200 expression. Hematoxylin-eosin staining was utilized to detect relative muscle area in gastrocnemius muscle. Electron microscopy was applied to observe changes in allograft ultrastructure. There was no obvious change in morphological appearance or ultrastructure, breaking force, or cytotoxicity of human acellular nerve allografts after cryopreservation at -80°C. Moreover, there was no remarkable change in neurofilament 200 expression, myelin sheath thickness, or muscle atrophy when fresh or cryopreserved rat acellular nerve allografts were applied to repair nerve injury in rats. These results suggest that cryopreservation can greatly extend the storage duration of acellular nerve tissue allografts without concomitant alteration of the physiochemical and biological properties of the engineered tissue to be used for transplantation.