Merck
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  • Decellularized peripheral nerve grafts by a modified protocol for repair of rat sciatic nerve injury.

Decellularized peripheral nerve grafts by a modified protocol for repair of rat sciatic nerve injury.

Neural regeneration research (2020-12-04)
Arash Zaminy, Sara Sayad-Fathi, Farshad Moharrami Kasmaie, Zohreh Jahromi, Adib Zendedel
摘要

Studies have shown that acellular nerve xenografts do not require immunosuppression and use of acellular nerve xenografts for repair of peripheral nerve injury is safe and effective. However, there is currently no widely accepted standard chemical decellularization method. The purpose of this study is to investigate the efficiency of bovine-derived nerves decellularized by the modified Hudson's protocol in the repair of rat sciatic nerve injury. In the modified Hudson's protocol, Triton X-200 was replaced by Triton X-100, and DNase and RNase were used to prepare accelular nerve xenografts. The efficiency of bovine-derived nerves decellularized by the modified Hudson's protocol was tested in vitro by hematoxylin & eosin, Alcian blue, Masson's trichrome, and Luxol fast blue staining, immunohistochemistry, and biochemical assays. The decellularization approach excluded cells, myelin, and axons of nerve xenografts, without affecting the organization of nerve xenografts. The decellularized nerve xenograft was used to bridge a 7 mm-long sciatic nerve defect to evaluate its efficiency in the repair of peripheral nerve injury. At 8 weeks after transplantation, sciatic function index in rats subjected to transplantation of acellular nerve xenograft was similar to that in rats undergoing transplantation of nerve allograft. Morphological analysis revealed that there were a large amount of regenerated myelinated axons in acellular nerve xenograft; the number of Schwann cells in the acellular nerve xenograft was similar to that in the nerve allograft. These findings suggest that acellular nerve xenografts prepared by the modified Hudson's protocol can be used for repair of peripheral nerve injury. This study was approved by the Research Ethics Committee, Research and Technology Chancellor of Guilan University of Medical Sciences, Iran (approval No. IR.GUMS.REC.1395.332) on February 11, 2017.

材料
货号
品牌
产品描述

Roche
DNase I, from bovine pancreas
Sigma-Aldrich
Triton X-100 溶液, BioUltra, for molecular biology, ~10% in H2O
Sigma-Aldrich
核糖核酸酶A 来源于牛胰腺, for molecular biology, ≥70 Kunitz units/mg protein, lyophilized
Sigma-Aldrich
SIGMAFAST DAB,含金属增强剂, tablet