Restoration of neuronal functions by outgrowths regenerating at ~1mm/d from the proximal stumps of severed peripheral nerves takes many weeks or months, if it occurs at all, especially after ablation of nerve segments. Distal segments of severed axons typically degenerate in 1–3 days. The purpose of this study was to show that Wallerian degeneration could be prevented or retarded and lost behavioral function restored following ablation of 0.5 – 1 cm segments of rat sciatic nerves in host animals. This is achieved using 0.8 – 1.1cm microsutured donor allografts treated with bioengineered solutions varying in ionic and polyethylene glycol (PEG) concentrations (modified PEG-fusion procedure), being careful not to stretch any portion of donor or host sciatic nerves. Our data show that PEG-fusion permanently restores axonal continuity within minutes as initially assessed by action potential conduction and intracellular diffusion of dye. Behavioral functions mediated by the sciatic nerve are largely restored within 2 – 4 wk as measured by the Sciatic Functional Index (SFI). Increased restoration of sciatic behavioral functions after ablating 0.5 – 1 cm segments is associated with greater numbers of viable myelinated axons within, and distal to, PEG-fused allografts. Many such viable myelinated axons are almost-certainly spared from Wallerian degeneration by PEG-fusion. PEG-fusion of donor allografts may produce a paradigm-shift in the treatment of peripheral nerve injuries.

Extracellular functions of the endoplasmic reticulum chaperone protein calreticulin (CRT) are emerging. Here we show novel roles for exogenous CRT in both cutaneous wound healing and diverse processes associated with repair. Compared with platelet-derived growth factor-BB-treated controls, topical application of CRT to porcine excisional wounds enhanced the rate of wound re-epithelialization. In both normal and steroid-impaired pigs, CRT increased granulation tissue formation. Immunohistochemical analyses of the wounds 5 and 10 days after injury revealed marked up-regulation of transforming growth factor-␤3 (a key regulator of wound healing), a threefold increase in macrophage influx, and an increase in the cellular proliferation of basal keratinocytes of the new epidermis and of cells of the neodermis. In vitro studies confirmed that CRT induced a greater than twofold increase in the cellular proliferation of primary human keratinocytes, fibroblasts, and microvascular endothelial cells (with 100 pg/ml, 100 ng/ ml, and 1.0 pg/ml, respectively). Moreover, using a scratch plate assay, CRT maximally induced the cellular migration of keratinocytes and fibroblasts (with 10 pg/ml and 1 ng/ml, respectively). In addition, CRT induced concentration-dependent migration of keratinocytes, fibroblasts macrophages, and monocytes in chamber assays. These in vitro bioactivities provide mechanistic support for the positive biological effects of CRT observed on both the epidermis and dermis of wounds in vivo, underscoring a significant role for

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Background Approximately 12% of operations for traumatic neuropathy are for patients with segmental nerve loss and less than 50% of these injuries obtain meaningful functional recovery. Polyethylene glycol (PEG) therapy has been shown to improve functional outcomes after nerve severance and we hypothesized this therapy could also benefit nerve autografting. Methods A segmental rat sciatic nerve injury model was used, whereby a 0.5 cm defect was repaired with an autograft using microsurgery. Experimental animals were treated with solutions containing methylene blue (MB) and PEG; control animals did not receive PEG. Compound Actions Potentials (CAPs) were recorded before nerve transection, after solution therapy, and at 72 hours postoperatively. The animals underwent behavioral testing at 24 and 72 hours postoperatively. After sacrifice, nerves were fixed, sectioned, and immunostained to allow for quantitative morphometric analysis. Results The introduction of hydrophilic polymers greatly improved morphological and functional recovery of rat sciatic axons at 1–3 days following nerve autografting. PEG therapy restored CAPs in all animals and CAPs were still present 72 hours postoperatively. No CAPS were detectable in control animals. Footfall asymmetry scores and sciatic functional index scores were significantly improved for PEG therapy group at all time points (p <0.05 and p<0.001; p <0.001 and p <0.01). Sensory and motor axon counts were increased distally in nerves treated with PEG compared to control (p = 0.0189 and p = 0.0032). Conclusions PEG therapy improves early physiologic function, behavioral outcomes, and distal axonal density after nerve autografting.