Sciatic Nerve Regeneration Research Articles

TPP ionically cross-linked chitosan/PLGA microspheres for the delivery of NGF for peripheral nerve system repair

To control the release of nerve growth factor (NGF) in the injured peripheral nerve, NGF-loaded chitosan/PLGA composite microspheres ionically cross-linked by tripolyphosphate (TPP/Chitosan/PLGA-NGF) were prepared. The encapsulation efficiency of NGF ranged from 83.4 ± 1.5 % to 72.1 ± 1.6 % with TPP concentrations from 1 % to 10 %. Zeta potential and FT-IR analyses together with confocal microscopy demonstrated that multiple NGF-loaded PLGA microspheres were embedded in chitosan matrix, the mean size of TPP/Chitosan/PLGA-NGF microspheres ranged from 40.2 ± 3.4 to 49.3 ± 3.1 μm. The increase of TPP concentration improved the network stability and decreased the swelling ratio, resulting in the decreased NGF release from 67.7 ± 1.2 % to 45.7 ± 0.8 % in 49 days. The sustained release of NGF could promote PC12 cells differentiation and neurite growth in vitro. Moreover, in comparison with NGF solution without microencapsulation, TPP/Chitosan/PLGA-NGF microspheres enhanced sciatic nerve regeneration and prevented gastrocnemius muscle atrophy in rats. These results demonstrate the feasibility of using TPP/Chitosan/PLGA-NGF microspheres for neural tissue repair.

Nogo-C Inhibits Peripheral Nerve Regeneration by Regulating Schwann Cell Apoptosis and Dedifferentiation.

While Nogo protein demonstrably inhibits nerve regeneration in the central nervous system (CNS), its effect on Schwann cells in peripheral nerve repair and regeneration following sciatic nerve injury remains unknown. In this research, We assessed the post-injury expression of Nogo-C in an experimental mouse model of sciatic nerve-crush injury. Nogo-C knockout (Nogo-C–/–) mouse was generated to observe the effect of Nogo-C on sciatic nerve regeneration, Schwann cell apoptosis, and myelin disintegration after nerve injury, and the effects of Nogo-C on apoptosis and dedifferentiation of Schwann cells were observed in vitro. We found that the expression of Nogo-C protein at the distal end of the injured sciatic nerve increased in wild type (WT) mice. Compared with the injured WT mice, the proportion of neuronal apoptosis was significantly diminished and the myelin clearance rate was significantly elevated in injured Nogo-C–/– mice; the number of nerve fibers regenerated and the degree of myelination were significantly elevated in Nogo-C–/– mice on Day 14 after injury. In addition, the recovery of motor function was significantly accelerated in the injured Nogo-C–/– mice. The overexpression of Nogo-C in primary Schwann cells using adenovirus-mediated gene transfer promoted Schwann cells apoptosis. Nogo-C significantly reduced the ratio of c-Jun/krox-20 expression, indicating its inhibition of Schwann cell dedifferentiation. Above all, we hold the view that the expression of Nogo-C increases following peripheral nerve injury to promote Schwann cell apoptosis and inhibit Schwann cell dedifferentiation, thereby inhibiting peripheral nerve regeneration.

Combined Use of Chitosan and Olfactory Mucosa Mesenchymal Stem/Stromal Cells to Promote Peripheral Nerve Regeneration In Vivo

Peripheral nerve injury remains a clinical challenge with severe physiological and functional consequences. Despite the existence of multiple possible therapeutic approaches, until now, there is no consensus regarding the advantages of each option or the best methodology in promoting nerve regeneration. Regenerative medicine is a promise to overcome this medical limitation, and in this work, chitosan nerve guide conduits and olfactory mucosa mesenchymal stem/stromal cells were applied in different therapeutic combinations to promote regeneration in sciatic nerves after neurotmesis injury. Over 20 weeks, the intervened animals were subjected to a regular functional assessment (determination of motor performance, nociception, and sciatic indexes), and after this period, they were evaluated kinematically and the sciatic nerves and cranial tibial muscles were evaluated stereologically and histomorphometrically, respectively. The results obtained allowed confirming the beneficial effects of using these therapeutic approaches. The use of chitosan NGCs and cells resulted in better motor performance, better sciatic indexes, and lower gait dysfunction after 20 weeks. The use of only NGGs demonstrated better nociceptive recoveries. The stereological evaluation of the sciatic nerve revealed identical values in the different parameters for all therapeutic groups. In the muscle histomorphometric evaluation, the groups treated with NGCs and cells showed results close to those of the group that received traditional sutures, the one with the best final values. The therapeutic combinations studied show promising outcomes and should be the target of new future works to overcome some irregularities found in the results and establish the combination of nerve guidance conduits and olfactory mucosa mesenchymal stem/stromal cells as viable options in the treatment of peripheral nerves after injury.

Long-Term Outcome of Sciatic Nerve Regeneration Using Bio3D Conduit Fabricated from Human Fibroblasts in a Rat Sciatic Nerve Model

Previously, we developed a Bio3D conduit fabricated from human fibroblasts and reported a significantly better outcome compared with artificial nerve conduit in the treatment of rat sciatic nerve defect. The purpose of this study is to investigate the long-term safety and nerve regeneration of Bio3D conduit compared with treatments using artificial nerve conduit and autologous nerve transplantation. We used 15 immunodeficient rats and randomly divided them into three groups treated with Bio3D (n = 5) conduit, silicon tube (n = 5), and autologous nerve transplantation (n = 5). We developed Bio3D conduits composed of human fibroblasts and bridged the 5 mm nerve gap created in the rat sciatic nerve. The same procedures were performed to bridge the 5 mm gap with a silicon tube. In the autologous nerve group, we removed the 5 mm sciatic nerve segment and transplanted it. We evaluated the nerve regeneration 24 weeks after surgery. Toe dragging was significantly better in the Bio3D group (0.20 ± 0.28) than in the silicon group (0.6 ± 0.24). The wet muscle weight ratios of the tibial anterior muscle of the Bio3D group (79.85% ± 5.47%) and the autologous nerve group (81.74% ± 2.83%) were significantly higher than that of the silicon group (66.99% ± 3.51%). The number of myelinated axons and mean myelinated axon diameter was significantly higher in the Bio3D group (14708 ± 302 and 5.52 ± 0.44 μm) and the autologous nerve group (14927 ± 5089 and 6.04 ± 0.85 μm) than the silicon group (7429 ± 1465 and 4.36 ± 0.21 μm). No tumors were observed in any of the rats in the Bio3D group at 24 weeks after surgery. The Bio3D group showed significantly better nerve regeneration and there was no significant difference between the Bio3D group and the nerve autograft group in all endpoints.

Results of a comparative valuation of the efficiency of using the plasmid construct pBud-VEGF165-FGF2 in models of autograft of the sciatic nerve defect and tubulation with the NeuraGen® collagen tube

Traumatic injuries of peripheral nerves lead to profound disability in patients with partial or total loss of limb function. There remains the question about the use of technologies for detecting defects of the peripheral nerve with concurrent of its regeneration. In the study it has been investigated the effect of the gene-therapeutic plasmid construct pBud-VEGF165-FGF2 with various methods of overcoming 5 mm diastasis of the sciatic nerve: nerve autograft and tubulation with the NeuraGen® tube. In the study groups, assessment of sciatic nerve regeneration was based on functional and morphometric parameters. Direct injection of plasmid pBud-VEGF165-FGF2 stimulates regeneration and restoration of motor function in both groups, but with different efficacy. Comparative analysis of nerve defect replacement in combination with direct gene therapy showed the most effective approach with autologous insertion replacement by comparison to the NeuraGen. Thus, on the basis of the obtained data, we can assert that nerve autograft of the peripheral nerve remains the "gold standard” and provides the best hope of research in combination with the use of various regeneration stimulants.

Effects of Platelet-Rich Fibrin/Collagen Membrane on Sciatic Nerve Regeneration.

Alternative treatment approaches to improve the regeneration ability of damaged peripheral nerves are currently under investigation. The aim of the current study was to evaluate the effects of leucocyte/platelet-rich fibrin (L-PRF) with or without a collagen membrane as a supporter on crushed sciatic nerve healing in a rat model. Recovery of motor function and electrophysiologic measurements were evaluated at 4 weeks postoperatively. The whole number of myelinated axons, peripheral nerve axon density, average nerve fiber diameter (μm), and G-ratio were analyzed and compered among the groups. Functional, electrophysiological, and histological evaluations showed no significant difference among the groups with the exception of the L-PRF with collagen membrane groups that showed relatively positive effects on the functional and histological nerve recovery. In addition, the collagen membrane with L-PRF can be effect in nerve regeneration.

Graphene-based conductive fibrous scaffold boosts sciatic nerve regeneration and functional recovery upon electrical stimulation

Peripheral nerve injury (PNI) is a very common consequence of traumatic injury and may cause serious disturbance in mediating functions in the human body. It's an urgent issue to promote the sufficient morphologic and functional regeneration of injured peripheral nerve. In this study, combination of graphene-based conductive fibrous scaffold (GCFS) and exogenous electrical stimulation (ES) were developed as an effective strategy to repair PNI, and the possible mechanisms underlying nerve recovery were further explored. The prepared GCFSs without or with 1.0 wt% graphene possessed electrical conductivity of 5.27 × 10−6 S/m and 3.12 S/m, respectively. The in vitro studies showed that ES could accelerate migration of rat mesenchymal stem cells (MSCs) seeded on the GCFS, promote the secretion of neurotrophic factors, and meanwhile up-regulate gene and protein expressions of neural markers. In vivo studies using a rat sciatic nerve injure model revealed that ES could significantly enhance sciatic nerve regeneration and functional recovery after implantation of GCFS nerve guidance conduit, and the daily ES exhibited repair-promoting capacity comparable to the gold standard autograft. Fundamentally, ES facilitated nerve regeneration maybe partly by the recruitment of endogenous MSCs and modulation of macrophage phenotypes. Taken together, GCFS combined with ES present a feasibility strategy to promote regeneration and functional recovery of the injured peripheral nerve.

Recovery of sciatic nerve with complete transection in rats treated with leuprolide acetate: A gonadotropin-releasing hormone agonist

It has been reported that the Gonadotropin-releasing hormone (GnRH) and its agonist leuprolide acetate (LA) can act as promoters of nerve regeneration. The aim of this study is to evaluate the effect of LA in a complete transection model. Sciatic nerve injury (SNI) was performed using a complete nerve transection and immediately repaired by epineural sutures. Rats were divided into three groups: SHAM, SNI treated with LA (SNI + LA) or saline solution (SNI + SS) for 5 weeks. Sciatic nerve regeneration was evaluated by kinematic gait analyzes, electrophysiological, morphological and biochemical tests. SNI + LA group had a functional recovery in kinematic gait, an increase in ankle angle value and a faster walking speed, compound muscle action potential amplitude, nerve conduction velocity (NCV). Furthermore, the number of myelinated axons and microtubule-associated protein 2 (MAP-2) expression were also higher compared to SS group. In conclusion, LA treatment improves of gait, walking speed, NCV, axons morphometry and MAP-2 expression in rats with sciatic nerve complete transection. These results suggest that LA can be a potential treatment for peripheral nerve injuries.

High energy photobiomodulation therapy in the early days of injury improves sciatic nerve regeneration in mice

Introduction: Different studies have evaluated the effects of electrophysical agents on regeneration after peripheral nerve injury. Among them, the most used in clinical and experimental research is photobiomodulation therapy (PBMT). Objective: To analyze the effect of standard energy (16.8 J) of PBMT on peripheral nerve regeneration, applied at different periods after sciatic nerve injury in mice. Methods: Thirty male Swiss mice were divided into six groups: naive; sham; control; LLLT-01 (660 nm, 16.8 J of total energy emitted in 1 day); LLLT-04 (660 nm, 4.2 J per day, 16.8 J of total energy emitted in 4 days); LLLT-28, (660 nm, 0.6 J per day, 16.8 J of total energy emitted over 28 days). The animals were evaluated using thermal hyperalgesia, Sciatic Functional Index (SFI), and Static Sciatic Index (SSI). Data were obtained at baseline and after 7, 14, 21, and 28 days after surgery. Results: For the SFI and SSI, all groups showed significant differences compared to the control group, and the LLLT-04 group presented the best results among those receiving PBMT. In the assessment of thermal hyperalgesia, there was a significant difference in the 14th day of evaluation in the LLLT-04 group. Conclusion: The application of 16.8 J was useful in sciatic nerve regeneration with an improvement of hyperalgesia, with higher efficacy when applied in four days (4.2 J/day).

Neutrophil peptide-1 promotes the repair of sciatic nerve injury through the expression of proteins related to nerve regeneration

ABSTRACT Objectives Small-molecule polypeptide neutrophil peptide 1 (NP-1) was reported to promote the regeneration of the sciatic nerve after denervation, but the mechanisms underlying this effect of NP-1 are unclear. Here, we established a Sprague-Dawley rat model of crush injury to study the effect of a single intermuscular injection of NP-1 on the repair of injured peripheral nerves and elucidate the possible underlying mechanism. Methods 39 rats were randomly selected to join this study and divided into the blank control group (normal group, n=9), experimental group (NP-1 group, n=15), and negative control group (NS group, n=15). The dynamic expression of cytokines in different groups of nerve tissues during Wallerian degeneration was observed using protein chips at different time points after injury. Recovery of injured nerves was determined based on the general condition, local gross morphology of the nerve suture site, sciatic nerve function index, neuroelectrophysiology, and osmic acid staining at 6 weeks after the surgery. The recovery of effector function was determined based on wet weight, hematoxylin-eosin staining, modified Gomori staining, and nicotinamide adenine dinucleotide-tetrazolium reductase staining at 6 weeks after the surgery. Results It was found that a single topical administration of NP-1 promoted sciatic nerve regeneration after crush injury and affected the expression of proteins related to neurotrophy, inflammation, cell chemotaxis, and cell generation pathways.

Improving sciatic nerve regeneration by using alginate/chitosan hydrogel containing berberine.

Peripheral nerve injuries are the common results of trauma that lead to pain and handicap in patients. Berberine due to its properties like antibiotic, immunostimulant, antitumor, antimotility, and positive effect on neurological disorders can be used to enhance peripheral nerve injuries. In this study, alginate/chitosan hydrogel containing different concentrations of berberine (0, 0.1, 1, 10% (w/v)) was created, evaluated, and applied as a scaffold for sciatic nerve regeneration. To prepare hydrogel, sodium alginate was dissolved in distilled water and cross-linked with CaCl2, and chitosan was dissolved in acetic acid and cross-linked with β-glycerol phosphate. The structure, release, swelling, weight loss, cytocompatibility, and hemocompatibility of the prepared hydrogels were assessed. The sciatic nerve crush was created in rats and fabricated hydrogels were injected, and functional analysis was used to evaluate their effectiveness. The results of physical characterization of the hydrogel indicated that the initial average pore size was about 39 μm and about 70% of the main weight of hydrogels was lost after incubation for 21 days and hemocompatibility of hydrogels was also confirmed. The MTT assay showed the cytocompatiblity of hydrogels and also indicated that berberine has dose-dependence effect on cell proliferation. The in vivo results showed the positive effect of berberine especially the hydrogel contained 1% of berberine on regeneration of sciatic nerve. Based on this study, Alg/Chit hydrogel can be applied as a treatment to heal peripheral nerve injuries. Graphical abstract.

Regeneration of Rat Sciatic Nerve Using PLGA Conduit Containing Rat ADSCs with Controlled Release of BDNF and Gold Nanoparticles.

Implantation of a nerve guidance conduit (NGC) carrying neuroprotective factors is promising for repairing peripheral nerve injury. Here, we developed a novel strategy for repairing peripheral nerve injury by gold nanoparticles (AuNPs) and brain-derived neurotrophic factor (BDNF)-encapsulated chitosan in laminin-coated nanofiberof Poly(l-lactide-co-glycolide) (PLGA) conduit and transplantation of rat adipose-derived stem cells (r-ADSCs) suspended in alginate. Then, the beneficial effect of AuNPs, BDNF, and r-ADSCs on nerve regeneration was evaluated in rat sciatic nerve transection model. In vivo experiments showed that the combination of AuNPs- and BDNF-encapsulated chitosan nanoparticles in laminin-coated nanofiberof PLGA conduit with r-ADSCs could synergistically facilitate nerve regeneration. Furthermore, the in vivo histology, immunohistochemistry, and behavioral results demonstrated that the AuNPs- and BDNF-encapsulated chitosan nanoparticles in NGC could significantly reinforce the repair performance of r-ADSCs, which may also contribute to the therapeutic outcome of the AuNPs, BDNF, and r-ADSCs strategies. In this study, we found that the combination of AuNPs and BDNF releases in NGC with r-ADSCs may represent a new potential strategy for peripheral nerve regeneration.

In Vivo Gene Delivery of STC2 Promotes Axon Regeneration in Sciatic Nerves.

Neurons are vulnerable to injury, and failure to activate self-protective systems after injury leads to neuronal death. However, sensory neurons in dorsal root ganglions (DRGs) mostly survive and regenerate their axons. To understand the mechanisms of the neuronal injury response, we analyzed the injury-responsive transcriptome and found that Stc2 is immediately upregulated after axotomy. Stc2 is required for axon regeneration in vivo and in vitro, indicating that Stc2 is a neuronal factor regulating axonal injury response. The application of the secreted stanniocalcin 2 to injured DRG neurons promotes regeneration. Stc2 thus represents a potential secretory protein with a feedback function regulating regeneration. Finally, the in vivo gene delivery of STC2 increases regenerative growth after injury in peripheral nerves in mice. These results suggest that Stc2 is an injury-responsive gene required for axon regeneration and a potential target for developing therapeutic applications.

Reconstruction of the Rat Sciatic Nerve by Using Biodegradable and Non-Biodegradable Conduits.

The aim of the study was to compare two types of conduits made of either non-resorbable Reperen or resorbable Tissucol for their effects on the regeneration of the rat sciatic nerve under conditions of stump diastasis.Materials and Methods.The experiments were carried out using outbred white male rats of the reproductive age (n=14). The animals were divided into three groups: group 1, intact (n=5), used for studying the morphology of the sciatic nerve; group 2 (n=4) — nerve plastic surgery was performed using a conduit made of non-resorbable Reperen; group 3 (n=5) — surgery was performed using a conduit made of resorbable Tissucol. The animals were anesthetized with isoflurane. After a complete transection of the sciatic nerve in the middle third of the thigh, its stumps were inserted into a conduit of an internal diameter of 2 mm and a length of 10 mm, filled with saline. Diastasis of 5 mm in length was created by spreading the nerve ends and securing the epineurium to the tube edges with 8/0 polypropylene sutures. A total count of myelinated nerve fibers was performed in the area of repair (tubulation) and the distal part of the nerve; the formation of connective tissue sheaths was assessed 14 weeks after the operation.Results.According to the morphological assessment, both types of conduits (resorbable and non-resorbable) caused the similar number of fibers to restore in the distal part of the repaired nerve; clinical characteristics of the animals in both groups were close to each other and to the norm.Conclusion.The results allow us to consider the conduit made of non-resorbable Reperen as a device promising for neuroplasty along with the resorbable conduit made of Tissucol.

Reduction in MLKL-mediated endosomal trafficking enhances the TRAIL-DR4/5 signal to increase cancer cell death

Mixed lineage kinase domain-like (MLKL) is an essential molecule of necroptosis, a cell death process that is initiated by direct disruption of the plasma membrane. During necroptosis, MLKL is phosphorylated by receptor interacting protein kinase-3 (RIPK3 or RIP3), and then translocates to the plasma membrane to disrupt membrane integrity. Recent data suggest that MLKL also has a RIP3-indendent function in the generation of intraluminal and extracellular vesicles (EVs), as well as in myelin sheath breakdown when promoting sciatic nerve regeneration. Here we show that depletion of MLKL enhances TRAIL-induced cell death in a RIP3-independent manner. Depletion of MLKL leads to prolonged cytotoxic signals that increase TRAIL-induced cell death. Initially, TRAIL binds to DR5 at the cell surface and is endocytosed at similar rates in MLKL-expressing and MLKL-depleted cells, eventual degradation of intracellular TRAIL by the lysosome is delayed in MLKL-depleted cells, corresponding with prolonged/enhanced intracellular signals such as p-ERK and p-p38 in these cells. Colocalization of TRAIL with the marker of early endosomes, EEA1 suggests that TRAIL is accumulated in early endosomes in MLKL-depleted cells compared to MLKL-expressing cells. This indicates that depletion of MLKL reduces receptor-ligand endosomal trafficking leading to increased TRAIL-cytotoxicity. An MLKL mutant that compromises its necroptotic function and its function in the generation of EVs was sufficient to rescue MLKL deficiency, suggesting that the N-terminal structural elements necessary for these functions are not required for the function of MLKL in the intracellular trafficking associated with regulating death receptor cytotoxicity. A reduction in MLKL expression in cancer cells would therefore be expected to result in enhanced TRAIL-induced therapeutic efficacy.

Human Wharton's Jelly Mesenchymal Stem Cell-Mediated Sciatic Nerve Recovery Is Associated with the Upregulation of Regulatory T Cells.

The acceleration of peripheral nerve regeneration is crucial for functional nerve recovery. Our previous study demonstrated that human Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSC) promote sciatic nerve recovery and regeneration via the direct upregulation and release of neurotrophic factors. However, the immunomodulatory role of hWJ-MSC in sciatic nerve recovery remains unclear. The effects of hWJ-MSC on innate immunity, represented by macrophages, natural killer cells, and dendritic cells, as well as on adaptive immunity, represented by CD4+ T, CD8+ T, B, and regulatory T cells (Tregs), were examined using flow cytometry. Interestingly, a significantly increased level of Tregs was detected in blood, lymph nodes (LNs), and nerve-infiltrating cells on POD7, 15, 21, and 35. Anti-inflammatory cytokines, such as IL-4 and IL-10, were significantly upregulated in the LNs and nerves of hWJ-MSC-treated mice. Treg depletion neutralized the improved effects of hWJ-MSC on sciatic nerve recovery. In contrast, Treg administration promoted the functional recovery of five-toe spread and gait stance. hWJ-MSC also expressed high levels of the anti-inflammatory cytokines TGF-β and IL-35. This study indicated that hWJ-MSC induce Treg development to modulate the balance between pro- and anti-inflammation at the injured sciatic nerve by secreting higher levels of anti-inflammatory cytokines.

Anisotropically Conductive Biodegradable Scaffold with Coaxially Aligned Carbon Nanotubes for Directional Regeneration of Peripheral Nerves.

Fascicular rearrangement of an injured peripheral nerve requires reconnection of nerve sprouts from anterior and Büngner bands from distal sides of the lesion, failing to which leads to inefficient regeneration of the injured nerve. However, existing neural scaffolds have limited neuroregeneration efficiency because of either the lack of alignment of fibers and a conductive second phase, leading to compromised electrical conductivity, or the lack of extracellular matrix components and in vivo validation. The present study reports a biocompatible, multiwall carbon nanotube (MWCNT)-reinforced, anisotropically conductive, electrospun, aligned nanofibrous scaffold, ensuring maximal peripheral nerve regeneration. Electrospinning parameters were modulated to deposit random and parallel fibers in separate scaffolds for comparative analysis on the effect of fiber alignment on regeneration. Both types of scaffolds were reinforced with MWCNTs to impart electrical conductivity. Nonreinforced scaffolds were nonconductive. In this comparative study, MWCNT-reinforced, aligned scaffolds showed better tensile property with increased conductivity along the direction of alignment, thereby ensuring an escalated neural-regeneration rate. Collectively, in vitro studies established the scaffolds to be highly biocompatible, promoting cell growth and proliferation. With 85% more anisotropic conductivity in the direction of the alignment and the degradation kinetics tuned to the regeneration regime, the MWCNT-reinforced, aligned scaffold efficiently healed injured sciatic nerves in rats within 30 days. Rigorous revivification of the tissue was due to coordinated Wallerian degeneration and expedited guided axonal regeneration. Structural and functional analysis of nerves in vivo showed the aligned, MWCNT-reinforced scaffold to be very efficient in peripheral sciatic nerve regeneration. This study notes the efficacy of the coaxially aligned, MWCNT-reinforced neural scaffold, with a capability of establishing remarkable advancement in the field of peripheral neural regeneration.

Grafts of human adipose-derived stem cells into a biodegradable poly (acid lactic) conduit enhances sciatic nerve regeneration

Despite the regenerative potential of the Peripheral Nervous System (PNS), injuries with loss of a nerve segment make the functional recovery a challenge. This work aimed to investigate the effects of the association of biodegradable conduits of poly (lactic acid) (PLA) with human adipose-derived stem cells (hADSCs) on the regeneration of the sciatic nerve. C57BL / 6 male mice were submitted to sciatic nerve transection followed by tubulization with PLA conduit. Animals were allocated in two groups: the first received an injection of DMEM inside the conduit (DMEM) and the second received hADSCs inside it (hADSC). Sensory and motor functions were assessed by the pinprick test and electroneuromiography, respectively. To assess neuronal survival the retrograde tracer fluorogold was injected into the sciatic nerve distally to the lesion site. One week after that, animals were sacrificed, tissues harvested and processed for morphological evaluation. After eight weeks, all animals showed sensory recovery in the pinprick test and there was no significant difference between the two groups. The amplitude of the compound muscle action potential was higher in the hADSCs group. The number of myelinated nerve fibers, muscle cells and motor plates was higher in the hADSC group. There was also greater survival of sensory and motor neurons in the hADSC animals. These results suggest that the association of PLA conduit and cell therapy with hADSCs leads to a better functional and morphological recovery after sciatic nerve transection.

Micro-grooved nerve guidance conduits combined with microfiber for rat sciatic nerve regeneration

In order to accelerate the regeneration of injured nerves, highly aligned structures that mimic native tissues are essential for nerve guidance conduits (NGC). In this study, a biodegradable porous micro-patterned NGCs were manufactured by use of capillary force lithography and salt leaching method, which filled as wrapped with aligned electrospun microfiber (PA-NGC) using. These combined NGC showed 5μm aligned groove pattern on the interior wall and were filled with 4–5μm align fibers for a fascicular structure to mimic the native nerve. The highly aligned structure has increased PC12 migration and morphological elongation compared with random microfiber. For the in vivo study, we bridged in the 10mm sciatic nerve defect margins with NGC. The PA-NGC were compared to flat NGCs filled with random (FR-NGC) or aligned fiber (FA-NGC) and patterned NGCs filled with random fiber (PR-NGC). The PA-NGC showed as dramatically re-bridged neurofilament and higher sciatic function index, onset-to-peak of amplitude, muscle weight and fiber diameter compared to other groups after 8 weeks of implantation. In our results, the PA-NGC not only can serve as a design for functional NGC without growth factor but also can be used in clinical application for peripheral nerve regeneration.

Evaluating effect of alginate/chitosan hydrogel containing 4-Methylcatechol on peripheral nerve regeneration in rat model

Some of the trauma patients (about 3%) suffer from peripheral nerve injuries and unfortunately, it is still a significant clinical problem to regenerate and functional recovery of them. These days, peripheral nerve damages have been treated with various methods and materials. The goal of the present study is to evaluate the effect of various amounts of 4-Methylcatechol (4-MC) on peripheral nerve regeneration. For this target, alginate/chitosan hydrogel was used to deliver different percentages (0.1%, 1%, and 10% (w/v)) of 4-MC to the sciatic nerve-injured sites in a rat model. Before that, the properties of hydrogels were assessed and after injecting, different functional analyses were used to evaluate the efficacy of fabricated hydrogels on peripheral nerve regeneration. Our results exhibited that the pore size of hydrogels was appropriate for cell proliferation (26–42 µm). Biodegradability of hydrogels was about 70% after 7 days and hemocompatibility of hydrogels was also confirmed. The MTT assay showed that the hydrogels not only do not have toxicity effects but also can improve the proliferation rate of PC12 cells. The functional results showed that 4-MC especially the group contained 10% of 4-MC enhance sciatic nerve regeneration. It could be concluded that fabricated hydrogels can use as suitable materials for peripheral nerve regeneration.