Mesenchymal stem cells in a polycaprolactone conduit enhance median-nerve regeneration, prevent decrease of creatine phosphokinase levels in muscle, and improve functional recovery in mice

doi:10.1016/j.neuroscience.2010.08.042

Neuroscience

Volume 170, Issue 4, 10 November 2010, Pages 1295-1303

https://doi.org/10.1016/j.neuroscience.2010.08.042 Get rights and content

Abstract

Although the majority of peripheral-nerve regeneration studies are carried out on the sciatic nerve, lesions of the upper extremities are more common in humans and usually lead to significant physical disabilities. The present study was driven by the hypothesis that a combination of strategies, namely grafts of mesenchymal stem cells (MSC) and resorbable polycaprolactone (PCL) conduits would improve median-nerve regeneration after transection. Mouse median nerves were transected and sutured to PCL tubes that were filled with either green fluorescent protein (GFP⁺) MSC in DMEM or with DMEM alone. During the post-operative period, animals were tested weekly for flexor digitorum muscle function by means of the grasping test. After 8 weeks, the proximal and middle portions of the PCL tube and the regenerating nerves were harvested and processed for light and electron microscopy. The flexor digitorum muscle was weighed and subjected to biochemical analysis for creatine phosphokinase (CK) levels. Scanning electron microscopy of the PCL tube 8 weeks after implantation showed clear signs of wall disintegration. MSC-treated animals showed significantly larger numbers of myelinated and unmyelinated nerve fibers and blood vessels compared with DMEM-treated animals. The flexor digitorum muscle CK levels were significantly higher in the MSC-treated animals, but muscle weight values did not differ between the groups. Compared with the DMEM-treated group, MSC-treated animals showed, by the grasping test, improved functional performance throughout the period analyzed. Immunofluorescence for S-100 and GFP showed, in a few cases, double-labeled cells, suggesting that transplanted cells may occasionally transdifferentiate into Schwann cells. Our data demonstrate that the polycaprolactone conduit filled with MSC is capable of significantly improving the median-nerve regeneration after a traumatic lesion.

Section snippets

Animals

For this study, we used 32 C57/Black6 mice, weighing between 25 and 30 g. The animals were housed in cages with food and water ad libitum and a 12/12-h light/dark cycle. All experiments were performed in accordance with the Ethics Committee for the Use of Experimental Animals of the Universidade Federal do Rio de Janeiro (Protocol DHE003). All efforts were taken in order to minimize number of animals used and their suffering.

The animals were divided into three groups according to the surgical

PCL in association with MSC is a suitable substrate for the regenerating nerve

We observed the ultrastructural features of the proximal and middle portions of the PCL conduit, prior to surgery (Fig. 1) and 8 and 12 weeks after surgery (Fig. 2) by scanning electron microscopy. We did not observe any tube fragments at the surgical site, or an exacerbated inflammatory response. The ultrastructural analysis of the PCL tube prior to implantation showed the presence of prominent pores on the outer surface (Fig. 1A′, B, B′), and the inner surface was smoother (Fig. 1A′, C, C′).

Discussion

Lesions in the nerves of the upper extremities are very common in humans, frequently leading to limiting and disabling pathologies, decreasing the quality of life of those affected. After a peripheral-nerve injury, there is an orchestrated sequence of events conducted by Schwann cells (trophic support), neurons (regenerative capacity), and the extracellular matrix (Boyd and Gordon, 2003, Gantus et al., 2006). Schwann cells are activated and proliferate, while immune cells are recruited to the

Acknowledgments

We are grateful to Jorge Luís da Silva for excellent technical assistance and to Klauss Mostacada for help with image processing. This study was supported by FAPERJ, CNPq, INNT-MCT, CAPES, and FUJB.

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Regeneration, Repair, and Developmental NeuroscienceResearch PaperMesenchymal stem cells in a polycaprolactone conduit enhance median-nerve regeneration, prevent decrease of creatine phosphokinase levels in muscle, and improve functional recovery in mice

Abstract

Section snippets

Animals

PCL in association with MSC is a suitable substrate for the regenerating nerve

Discussion

Acknowledgments

J Neurosci Methods

Exp Neurol

Semin Cell Dev Biol

Biomaterials

Exp Neurol

Int J Oral Maxillofac Surg

Exp Neurol

Brain Res

J Hand Surg Am

Hand Clin

Toxicon

Prog Neurobiol

J Clin Neurosci

Exp Neurol

Neuroscience

Int Rev Neurobiol

Biomaterials

J Neurosci Methods

Biocheml Biophys Res Commun

Remyelination of the spinal cord following intravenous delivery of bone marrow cells

Glia

Microvessels promote motor nerve survival and regeneration through local VEGF release following ectopic reattachment

Microcirculation

Regeneration, Repair, and Developmental Neuroscience
Research Paper
Mesenchymal stem cells in a polycaprolactone conduit enhance median-nerve regeneration, prevent decrease of creatine phosphokinase levels in muscle, and improve functional recovery in mice