Cellular migration and axonal outgrowth from adult mammalian peripheral nervesin vitro

The presence of contractile cells, their organization around regenerating nerve trunks, and the hypothetical effect of these organized structures on the extent of regeneration across a tubulated 10‐mm gap in the rat sciatic nerve were investigated. Collagen and silicone tubes were implanted both empty and filled with a collagen‐glycosaminoglycan (GAG) matrix. Nerves were retrieved at 6, 30, and 60 weeks postoperatively and time‐dependent values of the nerve trunk diameter along the tubulated length were recorded. The presence of myofibroblasts was identified immunohistochemically using a monoclonal antibody to α‐smooth muscle actin. Myofibroblasts were circumferentially arranged around the perimeter of regenerated nerve trunks, forming a capsule which was about 10 times thicker in silicone tubes than in collagen tubes. The nerve trunk diameter that formed inside collagen tubes was twice as large as that inside silicone tubes. In contrast, the collagen‐GAG matrix had a relatively small effect on capsule thickness or diameter of regenerate. It was hypothesized that the frequency of successful bridging by axons depends on the balance between two competitive forces: the axial forces generated by the outgrowth of axons and nonneuronal cells from the proximal stump and the constrictive, circumferential forces imposed by the contractile tissue capsule that promote closure of the wounded stumps and prevent axon elongation. Because the presence of the collagen‐GAG matrix has enhanced greatly the recovery of normal function of regenerates in silicone tubes, it was hypothesized that it accelerated axonal elongation sufficiently before the hypothetical forces constricting the nerve trunk in silicone tubes became sufficiently large. The combined data suggest a new mechanism for peripheral nerve regeneration along a tubulated gap. J. Comp. Neurol. 417:415–430, 2000. © 2000 Wiley‐Liss, Inc.

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Connective tissue response to tubular implants for peripheral nerve regeneration: The role of myofibroblasts

Chamberlain

et al. 2000

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Multidimensional long‐term time‐lapse microscopy of in vitro peripheral nerve regeneration

Öztürk¹,

Erdoğan²

2004

Microscopy Res & Technique

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In order to test the effectiveness of a new advanced time-lapse microscopy imaging and image processing and analysis system, and to do quantitative and qualitative temporal analyses of in vitro peripheral nerve regeneration, long-term time-lapse imaging of cultures of mouse dorsal root ganglia (DRGs) was performed. DRGs were placed in a Petri dish, covered with collagen gel, their attached peripheral nerves were cut in the middle, creating a gap, and the dish was filled with culture medium. Six preparations were kept on the time-lapse imaging system, which provides a suitable incubation environment and enables to capture images from multiple coordinates at x,y,z axes at desired time intervals for 13 days. In general, the time-lapse imaging system proved quite stable and efficient, although some improvements are certainly required. Two main components of peripheral nerve regeneration, outgrowth of axons and activities of resident cells, were examined. Axons started to grow during the first hour of incubation with a 16.5 microm/h rate and showed the slowest rates (0.7 microm/h) on days 8 and 9, after which they resumed higher speeds again. The first cell came out of the proximal end of the cut nerve on the second day and it was a Schwann cell (SC), which was the prominent cell type in the preparations throughout the experiment. SCs were higher in number (83.15% of all cells) but slower in migration (3.4 vs. 7.3 microm/h, P < 0.001) than other cells. Other observed characteristics of axonal outgrowth and cellular activity and interactions between axons and the cells are discussed.

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The Role of Endogenous Growth Factors to Support Corneal Endothelial Migration after Wounding in vitro

Schilling-Schön

Pleyer

Hartmann

et al. 2000

Experimental Eye Research

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Cellular migration and axonal outgrowth from adult mammalian peripheral nervesin vitro

Cited by 17 publications

References 60 publications

Connective tissue response to tubular implants for peripheral nerve regeneration: The role of myofibroblasts

Connective tissue response to tubular implants for peripheral nerve regeneration: The role of myofibroblasts

Multidimensional long‐term time‐lapse microscopy of in vitro peripheral nerve regeneration

The Role of Endogenous Growth Factors to Support Corneal Endothelial Migration after Wounding in vitro

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