Long-term in vivo regeneration of peripheral nerves through bioengineered nerve grafts

doi:10.1016/j.neuroscience.2011.02.052

Neuroscience

Volume 181, 5 May 2011, Pages 278-291

https://doi.org/10.1016/j.neuroscience.2011.02.052 Get rights and content

Abstract

Although autologous nerve graft is still the first choice strategy in nerve reconstruction, it has the severe disadvantage of the sacrifice of a functional nerve. Cell transplantation in a bioartificial conduit is an alternative strategy to improve nerve regeneration. Nerve fibrin conduits were seeded with various cell types: primary Schwann cells (SC), SC-like differentiated bone marrow-derived mesenchymal stem cells (dMSC), SC-like differentiated adipose-derived stem cells (dASC). Two further control groups were fibrin conduits without cells and autografts. Conduits were used to bridge a 1 cm rat sciatic nerve gap in a long term experiment (16 weeks). Functional and morphological properties of regenerated nerves were investigated. A reduction in muscle atrophy was observed in the autograft and in all cell-seeded groups, when compared with the empty fibrin conduits. SC showed significant improvement in axon myelination and average fiber diameter of the regenerated nerves. dASC were the most effective cell population in terms of improvement of axonal and fiber diameter, evoked potentials at the level of the gastrocnemius muscle and regeneration of motoneurons, similar to the autografts. Given these results and other advantages of adipose derived stem cells such as ease of harvest and relative abundance, dASC could be a clinically translatable route towards new methods to enhance peripheral nerve repair.

Highlights

▶Cell seeded fibrin conduits enhance nerve regeneration long term. ▶Cell seeded fibrin conduits reduce muscle atrophy following denervation. ▶Differentiated adipose stem cells are most effective population for nerve repair.

Section snippets

Experimental animals

All animal protocols were approved by the local veterinary commission in Lausanne, Switzerland and were carried out in accordance with the European Community Council directive 86/609/ECC for the care and use of laboratory animals. Male Sprague–Dawley rats (Janvier, France) weighing 250 g were used for this study.

Cell cultures and differentiation

All cells were obtained from Sprague–Dawley rats (Janvier, France). Schwann cells were isolated from sciatic nerves as previously described (di Summa et al., 2010) and maintained in

Post-operative complications and autotomy

All animals survived the surgical procedure and recovered from anaesthesia. One animal from the fibrin+SC group developed a postoperative subcutaneous hematoma at the surgical site which was evacuated with no consequences. No other surgical complications occurred except autotomy of the foot following sciatic nerve axotomy. Major autotomy of the experimental foot (considered as complete loss or damage to multiple phalanges and/or presence of open wounds), which lead to premature euthanasia of

Discussion

Successful crossing of a nerve gap depends on the formation of a new extracellular matrix scaffold, over which blood vessels, fibroblasts and Schwann cells can migrate and progress towards the distal nerve stump (Rodriguez et al., 2000). Indeed, surviving axons from the proximal stump will develop growth cones and extend along the connective strands bridging the gap. Unless axonal contact is re-established in a timely fashion, however, this growth supportive environment is not maintained (Walsh

Conclusion

In conclusion, SC-like differentiated adipose-derived stem cells confirmed in this long term in vivo experiment the neurotrophic potential expressed in vitro, closely following or even matching morphological and functional results of the autografts, the actual gold standard in clinical practice. These results suggest that SC-like differentiated ASC, may substitute SC and are likely to be one of the most clinically translatable cell types to be employed in nerve regeneration.

Acknowledgments

We are grateful to Prof Daniel Egloff for the excellent administrative support and to Dr. Dominique Schaakxs and Patricia Engels (Department of Plastic Surgery, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland) for help and assistance in animal experimental procedures and tissue processing. We are thankful to Prof. Damien Debatisse (Department of Neurosurgery, Universität Kliniken der Stadt, Köln, Germany) for advice about the neurophysiology procedures and Dr Lev Novikov and Dr

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Regeneration, Repair, and Developmental NeuroscienceResearch PaperLong-term in vivo regeneration of peripheral nerves through bioengineered nerve grafts

Abstract

Highlights

Section snippets

Experimental animals

Cell cultures and differentiation

Post-operative complications and autotomy

Discussion

Conclusion

Acknowledgments

Int J Biochem Cell Biol

Biomaterials

J Plast Reconstr Aesthet Surg

C R Biol

Exp Neurol

Neurosci Res

Mech Ageing Dev

Neurophysiol Clin

Exp Neurol

J Plast Reconstr Aesthet Surg

Eur J Cell Biol

Exp Neurol

J Physiol Paris

Brain Res

J Clin Neurosci

Exp Neurol

J Plast Reconstr Aesthet Surg

Neuroscience

Exp Neurol

Biochem Biophys Res Commun

Exp Neurol

Exp Neurol

Clin Neurophysiol

Neurosci Lett

Biomaterials

In vitro differentiation of human processed lipoaspirate cells into early neural progenitors

Plast Reconstr Surg

Peripheral nerve regeneration through guidance tubes

Neurol Res

Postnatal development of the adult pattern of motor axon distribution in rat muscle

Nature

Motor axons preferentially reinnervate motor pathways

J Neurosci

Phenotypic and functional characteristics of mesenchymal stem cells differentiated along a Schwann cell lineage

Glia

Tissue engineered nerve constructs: where do we stand?

J Cell Mol Med

Peripheral regeneration

Annu Rev Neurosci

What is the optimal value of the g-ratio for myelinated fibers in the rat CNS?A theoretical approach

PLoS One

Regeneration, Repair, and Developmental Neuroscience
Research Paper
Long-term in vivo regeneration of peripheral nerves through bioengineered nerve grafts