[Effect of pre-degeneration of peripheral nerves on plasticity of cultivated Schwann cells and their cell number in vitro].
Predegeneration of peripheral nerve grafts is known to improve axonal regeneration in the sciatic nerve of the rat. Predegeneration involves sectioning the donor nerve in situ for a period of time prior to harvesting in order to allow Wallerian degeneration to take place. It is suggested that proliferating Schwann cells are responsible for this regeneration-promoting effect due to an increased production and accumulation of trophic factors. The aim of this study was to evaluate whether the proliferation of Schwann cells is increased after various predegeneration periods and whether predegeneration does affect the production of trophic factors. The rat's sciatic nerve was transsected distal to the spinal ganglion. Predegeneration was allowed to take place for one, two, three and four days and one, two and three weeks, respectively. The nerve was resected and prepared for Schwann cell cultivation. Cells cultivated from the contralateral untreated nerve served as control. S100 immunostaining, nerve-growth factor (NGF), and N-cadherin were used to characterize Schwann cells. Viability was assessed by fluoresceine fluorescence staining. Proliferation index was determined by BrdU DNA incorporation. Cultivation of cells harvested from predegenerated nerves indicated an increased proliferation of Schwann cells compared to the control. Proliferation effects depended on the period of predegeneration. A higher cell yield was obtained as early as 24 hours and up to three weeks after predegeneration. Optimal proliferation was seen after one week of predegeneration. The rapid expansion of Schwann cell populations oppressed the development of contaminating fibroblasts. NGF and N-cadherin were expressed by all S100-positive cells. Predegeneration did not affect viability. The determined high mitotic activity of predegenerated Schwann cells in combination with an early onset of proliferation may explain the regenerating-promoting effect of predegenerated nerve grafts. The expression of both NGF and N-cadherin in the predegenerated cultures indicates a high level of plasticity with dedifferentiation capacity, which in turn promotes neural regeneration. Predegeneration allows a proliferation of Schwann cells without concomitant pharmacological treatment. Thus, predegeneration of peripheral nerves is considered to be a highly efficacious method if a high yield of activated Schwann cells is desired within a short period of time. This may become relevant in the future of repair of peripheral nerve lesions, when autologous Schwann cells with their capacity to provide neurotrophines and cell adhesion molecules are used as cellular prostheses to bridge nerve gaps.
Fansa
H
H
Klinik für Plastische, Wiederherstellungs- und Handchirurgie, Otto-von-Guericke-Universität Magdeburg.
Keilhoff
G
G
Frerichs
O
O
Wolf
G
G
Schneider
W
W
D000818:Animals; D001369:Axons / Q000502:physiology; D002452:Cell Count; D002455:Cell Division / Q000502:physiology*; D002478:Cells, Cultured; D008297:Male; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009416:Nerve Regeneration / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology*; D010525:Peripheral Nerves / Q000503:physiopathology*; D051381:Rats; D017208:Rats, Wistar; D012583:Schwann Cells / Q000502:physiology*; D012584:Sciatic Nerve / Q000503:physiopathology; D014855:Wallerian Degeneration / Q000503:physiopathology*