A Novel Model for Acute Peripheral Nerve Injury in the Horse and Evaluation of the Effect of Mesenchymal Stromal Cells Applied In Situ on Nerve Regeneration: A Preliminary Study

Abstract

Transplantation of mesenchymal stromal cells (MSCs) to sites of experimentally created nerve injury in laboratory animals has shown promising results in restoring nerve function. This approach for nerve regeneration has not been reported in horses. In this study, we first evaluated the in vitro ability of equine bone marrow-derived MSCs (EBM-MSCs) to trans-differentiate into Schwann-like cells and subsequently tested the MSCs in vivo for their potential to regenerate a transected nerve after implantation. The EBM-MSCs from three equine donors were differentiated into SCLs for 7 days, in vitro, in the presence of specialized differentiation medium and evaluated for morphological characteristics, by using confocal microscopy, and for protein characteristics, by using selected Schwann cell markers (GFAP and S100b). The EBM-MSCs were then implanted into the fascia surrounding the ramus communicans of one fore limb of three healthy horses after a portion of this nerve was excised. The excised portion of the nerve was examined histologically at the time of transection, and stumps of the nerve were examined histologically at day 45 after transplantation. The EBM-MSCs from all donors demonstrated morphological and protein characteristics of those of Schwann cells 7 days after differentiation. Nerves implanted with EBM-MSCs after nerve transection did not show evidence of nerve regeneration at day 45. Examination of peripheral nerves collected 45 days after injury and stem cell treatment revealed no histological differences between nerves treated with MSCs and those treated with isotonic saline solution (controls). The optimal delivery of MSCs and the model suitable to study the efficacy of MSCs in nerve regeneration should be investigated.

Regenerative Strategies in Treatment of Peripheral Nerve Injuries in Different Animal Models

“… 1-No horses exhibit any form of disability after the operation. [ 88 ] 2-SLCS from EBW-MSCS success in the expression of S-100b and GFAP but undifferentiated control cells express β 3 tubulin and GFAP and failed to express S-100b “neural progenitor markers”. RFP-BMSCs “red fluorescent protein-bone marrow stem cells”, p-MAPK “protein- mitogen-activated protein kinase”, p-ERK “protein-extracellular signal-regulated kinases”, CMAP “compound muscle action potential”, TENG “tissue engineered nerve graft”, NF “neurofilament”, MCV “muscle conduction velocity”, SF-based “silk fibroin-based”, FG-labeled “fluorogold-labeled”, DRGs “dorsal root ganglions”, SLCS from EBW-MSCS “Schwann-like cells from equine bone marrow-mesenchymal stem cells”, GFAP “Glial fibrillary acidic protein”, β 3 tubulin “class 3 tubulin protein”.…”

The Usefulness of Mesenchymal Stem Cells beyond the Musculoskeletal System in Horses

“…Horses suffer injury to peripheral nerves from trauma, metabolic and genetic disorders, toxins or degenerative and infectious diseases [ 126 ]. The degree of restoration of nerve function depends on the severity and chronicity of the damage, with the worst prognosis in cases where the nerve is transected [ 127 ]. There are few surgical techniques for repairing nerves, and clinical results are often poor [ 128 ].…”

“…In an in vivo model of acute peripheral nerve injury using three horses, allogeneic BM-MSCs were implanted into the fascia surrounding the ramus communicans of one forelimb after a portion was transected. No evidence of nerve regeneration was observed, neither were histological differences between MSC-treated and control injuries found 45 days later [ 127 ]. Nevertheless, improvement of nerve regeneration after MSC treatment has been observed in other large animals such as sheep [ 131 ].…”