Modification of Schwann cell gene expression by electroporation in vivo.
Clinical outcomes of nerve grafting are often inferior to those of end-to-end nerve repair. This may be due, in part, to the routine use of cutaneous nerve to support motor axon regeneration. In previous work, we have demonstrated that Schwann cells express distinct sensory and motor phenotypes, and that these promote regeneration in a modality-specific fashion. Intra-operative modification of graft Schwann cell phenotype might therefore improve clinical outcomes. This paper demonstrates the feasibility of electroporating genes into intact nerve to modify Schwann cell gene expression. Initial trials established 70 V, 5 ms as optimum electroporation parameters. Intact, denervated, and reinnervated rat tibial nerves were electroporated with the YFP gene and evaluated serially by counting S-100 positive cells that expressed YFP. In intact nerve, a mean of 28% of Schwann cells expressed the gene at 3 days, falling to 20% at 7 days with little expression at later times. There were no significant differences among the three groups at each time period. Electronmicroscopic evaluation of treated, intact nerve revealed only occasional demyelination and axon degeneration. Intra-operative electroporation of nerve graft is thus a practical means of altering Schwann cell gene expression without the risks inherent in viral transfection.
Department of Orthopaedic Surgery, Johns Hopkins University, 601 North Caroline Street, Baltimore, MD 21287, United States.
Manuela
M
Aspalter
Alka
A
Vyas
Jeffrey
J
Feiner
John
J
Griffin
Thomas
T
Brushart
Richard
R
Redett
D000818:Animals; D003714:Denervation; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D005260:Female; D015870:Gene Expression / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D008164:Luminescent Proteins / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D009416:Nerve Regeneration / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000502:physiology* / Q000637:transplantation; D013979:Tibial Nerve / Q000166:cytology / Q000502:physiology / Q000637:transplantation; D013997:Time Factors; D014162:Transfection / Q000379:methods
D008164:Luminescent Proteins