American Society for Peripheral Nerve

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Decellularized Vascularized Nerve Scaffolds For The Repair Of Peripheral Nerves
Tsering Wütrich, MSc1; Ioana Lese, MD2; Jérôme Duisit, MD3; Louis Maistriaux, MD3; Ekkehard Hewer, MD, PhD1; Cedric Zubler, cand med1; Ruslan Hluschchuk, MD, PhD1; Pierre Gianello, MD3; Benoit Lengele, MD, PhD3; Esther Voegelin, MD4; Robert Rieben, PhD1; Radu Olariu, MD2; Adriano Taddeo, PhD1
1University of Bern, Bern, Switzerland, 2Inselspital University Hospital of Bern, Bern, Switzerland, 3Université catholique de Louvain, Louvain, Belgium, 4Department of Plastic and Hand Surgery, Inselspital University Hospital of Bern, Bern, Switzerland

Peripheral nerve injuries are devastating, life-altering injuries. When they result in large nerve gaps (>6 cm) conventional nerve grafting is unreliable because of failure of centripetal revascularization. Vascularized autologous nerve grafts have shown promising results in these cases, however their availability is extremely limited. Decellularized scaffolds, which act as guide-conduits for the regenerating nerves, represent a promising approach to increase availability of nerve grafts and effective nerve repair. Here we report the development of a vascularized nerve scaffold in which the preserved vascular tree can be used for perfusion and recellularization in order to improve cell engraftment, supply of nutrient and, thus, regeneration and functional recovery.

Seven vascularized porcine sciatic nerve grafts were harvested and perfusion-decellularization was applied using a SDS/polar solvent protocol resulting in decellularized vascularized porcine sciatic nerve scaffolds (VPNS), which were characterized by analyzing cell and DNA content and preservation of extracellular matrix (ECM), vascular tree and cytokine content. Reendothelialization of decellularized scaffolds was conducted with porcine-aorta endothelial cells (PAEC) in a perfusion-bioreactor.

Scaffold vascularization was confirmed by angiography before and after decellularization. Morphologic examination of decellularized VPNS and analysis of the DNA content demonstrated cell and antigen clearance. ECM content and structures of the nerve fascicles were preserved. The VPNS's vasculature was characterized in 3D by using micro-computed tomography (microCT) imaging and showed optimal vasculature preservation down to the capillary level. Cytokines quantification demonstrated a strong preservation of growth factors and reduced preservation of pro-inflammatory cytokines. Reendothelialization experiments showed PAEC viability and engraftment with repopulation of the scaffold'svessels.

This study shows that perfusion-decellularization can be used to generate vascularized nerve scaffolds with preserved ECM structure and a functional vascular tree, which can be reendothelialized in vitro. As compared to non-vascularized conduits, vascularized engineered nerve scaffolds may represent an ideal approach for promoting better nerve regeneration in reconstructive surgery.

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