Promoting Nerve Regeneration in a Neurotmesis Rat Model Using Poly(DL-lactide-ε-caprolactone) Membranes and Mesenchymal Stem Cells from the Wharton’s Jelly:In VitroandIn VivoAnalysis

Abstract

In peripheral nerves MSCs can modulate Wallerian degeneration and the overall regenerative response by acting through paracrine mechanisms directly on regenerating axons or upon the nerve-supporting Schwann cells. In the present study, the effect of human MSCs from Wharton's jelly (HMSCs), differentiated into neuroglial-like cells associated to poly (DL-lactide-ε-caprolactone) membrane, on nerve regeneration, was evaluated in the neurotmesis injury rat sciatic nerve model. Results in vitro showed successful differentiation of HMSCs into neuroglial-like cells, characterized by expression of specific neuroglial markers confirmed by immunocytochemistry and by RT-PCR and qPCR targeting specific genes expressed. In vivo testing evaluated during the healing period of 20 weeks, showed no evident positive effect of HMSCs or neuroglial-like cell enrichment at the sciatic nerve repair site on most of the functional and nerve morphometric predictors of nerve regeneration although the nociception function was almost normal. EPT on the other hand, recovered significantly better after HMSCs enriched membrane employment, to values of residual functional impairment compared to other treated groups. When the neurotmesis injury can be surgically reconstructed with an end-to-end suture or by grafting, the addition of a PLC membrane associated with HMSCs seems to bring significant advantage, especially concerning the motor function recovery.

Evaluation of biodegradable electric conductive tube-guides and mesenchymal stem cells

“…The MSCs lot used in the present experimental study was characterized by flow cytometry analysis for a comprehensive panel of markers by PromoCell as previously reported [18,25] . The MSCs exhibited a mesenchymal-like shape with a flat and polygonal morphology during cell culture expansion.…”

“…MSCs karyotype by cytogenetic analysis: Cytogenetic analysis was carried out before in vivo application between passages 4 and 5 as previously reported [18,25] . Chromosome analysis was performed by one scorer on 20 Giemsa-stained metaphases and each MSC cell was scored for chromosome number.…”

“…To assess the nociceptive WRL, the hotplate test described by Masters et al [32] was used with some modifications and previously described by the authors [3,9,17,25,31,33] . Also, it is considered by the scientific bibliography that rats without sciatic nerve injury withdraw their paws from the hotplate within 4.3 s or less [9,17,31,33] .…”

“…The inflammatory modulation includes the Wallerian degeneration, a crucial step for nerve regeneration after neurotmesis injury [3] . Therefore, the use of cellular systems is a rational approach for promoting nerve by MSCs are primarily responsible for their therapeutic action, so it is particularly important to understand and fully characterize the MSCs secretome [18,24,25] . Recent studies demonstrated that MSCs produce and secrete multiple paracrine factors like interleukine-2 (IL-2), IL-6, IL-8, IL-12, IL-15, monocyte chemoattractant protein-1, macrophage inflammatory protein-1β, regulated on activation, normal T cell expressed and secreted, and platelet-derived growth factor-AA.…”

Olfactory Mucosa Mesenchymal Stem Cells and Biomaterials: A New Combination to Regenerative Therapies after Peripheral Nerve Injury

“…In contrast, high rates of acidic degradation and their products, rapid changes in mechanical properties and low solubility are important limitations [59]. PCL biomaterials are characterized by being transparent, which brings great clinical advantages during surgical application across the nerve gap defect [60], besides demonstrating eicacy in cases of large gaps [44]. Among the limitations, it is worth mentioning its high rigidity, which requires immersion in saline solution prior to use.…”