Floor plan - 2013 Annual Meeting - American Association for Hand ...

ASPN SCIENTIFIC PAPER SESSION A
Maintainance of Neuronal Differentiated Adipose-derived Stem Cells in Long Term Culture
Institution where the work was prepared: University of California, Irvine Medical Center, Orange, CA, USA
Suraj Kachgal, MS; Sanjay Dhar, PhD; Eul Sik Yoon, MD; Gregory R.D. Evans, MD; University of California, Irvine
INTRODUCTION:
Adipose-derived stem cells (AdSCs) have documented great potential to differentiate into cells of a neural phenotype. These cells provide
a great source for autologus transplantation into in vivo models of peripheral nervous system disorders. The present study investigates
the efficacy of a new neuronal induction media and whether it can maintain human AdSCs in a differentiated state in vitro for a
period of time that would correspond with nerve regeneration in vivo.
METHODS:
Human lipoaspirate was processed by standard methodologies and AdSCs from the product were extracted into culture. AdSCs were
briefly expanded in control medium and then subjected to culture in our neural induction media (DE-1) for periods of 1 day, 1, 2, 4, 6,
and 8 weeks. Cultures were probed for expression of neural-specific markers: NeuN, nestin, GFAP, vimentin, NSE, trk-A, and MAP2 via
immunocytochemistry, RT-PCR, and Western blot.
RESULTS:
Immunocytochemical staining of the neural-induced cells was positive for the markers GFAP, trk-A, nestin, and NeuN. Western blot
analysis revealed expression of early neural markers NSE and NeuN was found in control AdSCs and showed decreasing expression in
our neural-induced AdSCs, suggestive of a developing neural phenotype. Expression of the early glial marker vimentin was not present
in the control blot but was expressed in neural-induced AdSCs at day one. Vimentin expression tapered off to zero by week eight
while expression of the mature astrocyte marker GFAP expressed from day one to week eight. RT-PCR results indicate that all markers
except trk-A are transcribed in control and experimental groups, but Western blot analysis shows not all are transcribed.
CONCLUSION:
We have successfully established a medium which promotes neural differentiation of AdSCs and holds them in the differentiated state
for a period of time longer than previously reported. The media was successful in promoting the development of cells of a glial phenotype
as shown by expression profiles of vimentin and GFAP.
Repair of Partial Nerve Injury by Bypass Nerve Grafting with End-to-side Neurorrhaphy
Institution where the work was prepared: University of Mississippi Medical Center, Jackson, MS, USA
Tanya M. Oswald, MD; Feng Zhang; William C Lineaweaver; University of Mississippi Medical Center
BACKGROUND:
The peripheral nerve injury without disruption of the anatomic continuity of the nerve often results in formation of neuromas-in-continuity.
Management of this partial nerve injury is notoriously difficult. The purpose of this study was to determine the efficacy of bypass
nerve grafting with end-to-side neurorrhaphy in repair of partial nerve injury in a rabbit model.
METHODS:
Thirty-six adult male New Zealand rabbits were divided into three groups. The partial nerve injury was created by removal of a segment
of the lateral fascicle of the left peroneal nerve. In Group one, the injured nerve was repaired with a nerve graft bypassing the injury site
in an end-to-side fashion 4 weeks after injury. In Group two, the injured nerve was repaired with an end-to-end interposition nerve grafting
6 weeks after injury. The injured nerve without repair was used as the control. At the 16th week after nerve repair in groups one and
two, and 20 weeks after the initial nerve injury in the control group, the nerves were dissected for electrophysiological examination and
biopsied for histology and molecular marker expressions.
RESULTS:
The nerve repair with interposition nerve grafting achieved maximal functional recovery. However, the motor nerve conduction velocity
(MCV) and compound motor action potential (CMAP) in nerve repair with the bypass nerve grafting were significantly higher than
that in the nerve injury without repair. Histologically, the regenerated myelinated axons and unmyelinated axons were present in the
distal peroneal nerves in the bypass nerve grafts. The axon counts in nerve repair with bypass nerve grafting were also significantly higher
than that in the nerve injury without repair. The comparisons of the ciliary neurotrophic factor (CNTF) and the calcitonin gene related
peptide (CGRP) gene expressions between nerves with and without repair were significantly different.
CONCLUSION:
End-to-side bypass nerve grafting can significantly improve the functional recovery in the nerve with partial injury and may be a useful
repair strategy in neuroma-in-continuity.
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