American Society for Peripheral Nerve

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Human Epineural Sheath Conduits Supported with Human Mesenchymal Stem Cells Enhances Nerve Regeneration in Rat Experimental Model
Marcin Strojny, MD; Joanna Cwykiel, MS; Maria Siemionow, MD PhD DSc
University of Illinois at Chicago, Chicago, IL

INTRODUCTION: The disadvantages of nerve autografts, the gold standard of peripheral nerve regeneration, such as neuroma formation, scarring and limited availability yields the need for an alternative method of peripheral nerve management. Nerve allografts offer an unlimited source of nerve tissue, which can be modified to the recipient's nerve diameter to support nerve recovery after trauma. This study assessed the effect of human Epineural Sheath Conduit (hESC) supported with Human Mesenchymal Stem Cells (hMSC) on restoration of 20mm long nerve defect in a nude rat model.

METHODS: Restoration of 20mm sciatic nerve defect with hESC created from human sciatic nerve supported with hMSC was tested in 4 experimental groups: Group 1: no repair control (n=6), Group 2: autograft control (n=6), Group 3: hESC (n=6), Group 4: hESC + hMSC (n=6). Assessment for weight loss, limb autocannibalization or hind limb pressure sores, was performed daily on each rat. Any animal displaying these symptoms was disqualified from the study. Functional tests of toe-spread and pinprick were performed at 1, 3, 6, 9, 12 weeks after repair. At 12 weeks, nerve samples were collected for immunostaining of HLA-DR, VEGF, NGF, laminin B. Muscle samples were gathered for Gastrocnemius Muscle Index (GMI) and muscle fiber area ratio measurements.

RESULTS: Daily Clinical Assessment did not disqualify any of the animals from the study. Macroscopic evaluation of epineural conduit repair site at 12 weeks showed preservation of a normal nerve shape without tissue adhesion or local signs of inflammation and good vascularization was confirmed in all groups. The best sensory and motor recovery was observed in Group 2, followed by Group 4, Group 3 and Group 1 (pinprick 3.0 vs. 2.33 vs. 2.0 vs. 0.5; toe-spread 1.83 vs. 1.5 vs. 1.0 vs. 0.13, respectively). GMI and muscle fiber area ratio was the highest for Group 2 (0.322/0.414) followed by Group 4 (0.285/0.306), Group 3 (0.274/0.286) and Group 1 (0.167/0.098). Immunostaining for expression of VEGF, NGF and laminin B was highest for Group 2, followed by Group 4, Group 3 and Group 1. HLA-DR was expressed in Group 4.

CONCLUSIONS: We confirmed the feasibility of human ESC creation. Efficacy of hESC conduit in nerve regeneration was proved at 12 weeks by functional tests and cytokines expression comparable with autograft control. Our novel Human ESC provides a potential alternative to the autograft nerve repair. The role of local hMSC application in nerve regeneration needs to be further established.

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