Sutureless nerve repair has been regarded as a promising technique for nerve repair as the suture materials often results in neuroma formation and scar tissue that impede nerve regeneration. The aim of this study was to analyze the mechanical stability and morphological outcome of sutureless repair using fibrin glue conduit and an alternative approach of modified suture placement. Using rat sciatic nerve, we tested the following experimental conditions: conventional suture repair; single suture combined with fibrin glue repair, and fibrin conduit reinforced with modified suture or fibrin glue. Nerve detachment anatomical measures such as axon density, myelin, and fiber caliber were analyzed for evaluation of nerve regeneration. Muscle atrophy were evaluated by muscle wet weight and H&E staining. All animals in sutureless repair group exhibited complete detachment or elongation by two or four weeks after repair. No detachment was found in any other groups. Animals treated with fibrin conduit reinforced with modified suture showed better axonal regeneration with good alignment. There were no significant differences in axon caliber among the groups. Muscle atrophy was found in all groups and there was no significant difference in muscle wet-weight among the groups. In summary, sutureless nerve repair with fibrin glue was mechanically unstable for resistance of mechanical stretches, fibrin glue conduit with modified suture placement is mechanically stable and resulted in better morphological outcome.

Peripheral nerve injuries often result in lifelong disabilities despite advanced surgical interventions, indicating the urgent clinical need for effective therapies. In order to improve the potency of adipose-derived stem cells (ASC) for nerve regeneration, the present study focused primarily on ex-vivo stimulation of ASC by using growth factors, i.e., nerve growth factor (NGF) or vascular endothelial growth factor (VEGF) and secondly on fibrin-hydrogel nerve conduits (FNC) assisted ASC delivery strategies, i.e., intramural vs. intraluminal loading. ASC were stimulated by NGF or VEGF for 3 days and the resulting secretome was subsequently evaluated in an in vitro axonal outgrowth assay. For the animal study, a 10 mm sciatic nerve gap-injury was created in rats and reconstructed using FNC loaded with ASC. Secretome derived from NGF-stimulated ASC promoted significant axonal outgrowth from the DRG-explants in comparison to all other conditions. Thus, NGF-stimulated ASC were further investigated in animals and found to enhance early nerve regeneration as evidenced by the increased number of β-Tubulin III+ axons. Notably, FNC assisted intramural delivery enabled the improvement of ASC’s therapeutic efficacy in comparison to the intraluminal delivery system. Thus, ex-vivo stimulation of ASC by NGF and FNC assisted intramural delivery may offer new options for developing effective therapies.

We report on a potential strategy involving the exogenous neurotrophic factors (NTF) for enhancing the neurotrophic capacity of human adipose stem cells (ASC) in vitro. For this, ASC were stimulated for three days using NTF, i.e., nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT3), NT4, glial cell-derived neurotrophic factor (GDNF), and ciliary neurotrophic factor (CNTF). The resulting conditioned medium (CM) as well as individual NTF exhibited distinct effects on axonal outgrowth from dorsal root ganglion (DRG) explants. In particular, CM derived from NT3-stimulated ASC (CM-NT3-ASC) promoted robust axonal outgrowth. Subsequent transcriptional analysis of DRG cultures in response to CM-NT3-ASC displayed significant upregulation of STAT-3 and GAP-43. In addition, phosphoproteomic analysis of NT3-stimulated ASC revealed significant changes in the phosphorylation state of different proteins that are involved in cytokine release, growth factors signaling, stem cell maintenance, and differentiation. Furthermore, DRG cultures treated with CM-NT3-ASC exhibited significant changes in the phosphorylation levels of proteins involved in tubulin and actin cytoskeletal pathways, which are crucial for axonal growth and elongation. Thus, the results obtained at the transcriptional, proteomic, and cellular level reveal significant changes in the neurotrophic capacity of ASC following NT3 stimulation and provide new options for improving the axonal growth-promoting potential of ASC in vitro.