Direct gene transfer into rabbit peripheral nerve in vivo.

Abstract

Exogenous gene suture was used to achieve peripheral nerve anastomoses to probe into the feasibility that the sites of anastomoses of nerves directly transfer gene and thus enable gene to be expressed at the sites of anastomoses under the condition that perfect nerve anastomoses are ensured. PCMV beta plasmid containing cytomegalovirus promoter (CMV promoter) and Escherichia coli (E. coli) beta-Galactosidase (beta-Gal) structural gene (lacZ gene) was conducted. A soaked medical 8-0nylon suture was used to perform epineurial repair of rabbit sciatic nerve. In the control group a suture soaked in sucrose PBS was used, while in the experimental group a suture soaked in PCMV beta plasmid solution was applied. The sites of anastomoses of nerves by stages were taken out, and beta-Gal histochemical staining was performed and beta-Gal enzyme activity was assayed with 5-bromo-4-chloro-3-indolyl-beta-D-galactoside. Results showed that the sites of anastomoses of nerves were taken out 2 days, 7 days, 14 days and 30 days respectively after the operation. The beta-Gal histochemical stains at the sites of anastomoses showed no indigo positive cells at different stages in the control group, whereas displayed indigo positive cells in the experimental group. In the control group, no beta-Gal enzyme activity was detected at different stages after operation, but in the experimental group, beta-Gal enzyme activity could be detected from the 3rd day to the 30th day after operation. It was concluded that by using exogenous gene suture, exogenous gene could be transferred to the sites of peripheral nerve and expressed the exogenous gene expression products with bioactivity, which provided the feasibility of using gene therapy to accelerate the recovery of nerve function.