Introducing human adipose-derived mesenchymal stem cells to Avance^Ⓡ nerve grafts and NeuraGen^Ⓡ nerve guides

Background: When direct nerve coaptation is impossible after peripheral nerve injury, autografts, processed allografts, or conduits are used to bridge the nerve gap. The purpose of this study was to examine if human adipose-derived Mesenchymal Stromal/Stem Cells (MSCs) could be introduced to commercially available nerve graft substitutes and to determine cell distribution and the seeding efficiency of a dynamic seeding strategy. Methods: MTS assays examined the viability of human MSCs after introduction to the Avance^Ⓡ Nerve Graft and the NeuraGen^Ⓡ Nerve Guide. MSCs were dynamically seeded on nerve substitutes for either 6, 12, or 24 h. Cell counts, live/dead stains, Hoechst stains, and Scanning Electron Microscopy (SEM) revealed the seeding efficiency and the distribution of MSCs after seeding. Results: The viability of MSCs was not affected by nerve substitutes. Dynamic seeding led to uniformly distributed MSCs over the surface of both nerve substitutes and revealed MSCs on the inner surface of the NeuraGen^Ⓡ Nerve Guides. The maximal seeding efficiency of NeuraGen^Ⓡ Nerve Guides (94%), obtained after 12 h was significantly higher than that of Avance^Ⓡ Nerve Grafts (66%) (p = 0.010). Conclusion: Human MSCs can be dynamically seeded on Avance^Ⓡ Nerve Grafts and NeuraGen^Ⓡ Nerve Guides. The optimal seeding duration was 12 h. MSCs were distributed in a uniform fashion on exposed surfaces. This study demonstrates that human MSCs can be effectively and efficiently seeded onto commercially available nerve autograft substitutes in a timely fashion and sets the stage for the clinical application of MSC-seeded nerve graft substitutes clinically.