Repair Of Peripheral Nerve Defects Research Articles

Chitosan tubes enriched with fresh skeletal muscle fibers for delayed repair of peripheral nerve defects

Nerve regeneration after delayed nerve repair is often unsuccessful. Indeed, the expression of genes associated with regeneration, including neurotrophic and gliotrophic factors, is drastically reduced in the distal stump of chronically transected nerves; moreover, Schwann cells undergo atrophy, losing their ability to sustain regeneration. In the present study, to provide a three-dimensional environment and trophic factors supporting Schwann cell activity and axon re-growth, we combined the use of an effective conduit (a chitosan tube) with a promising intraluminal structure (fresh longitudinal skeletal muscle fibers). This enriched conduit was used to repair a 10-mm rat median nerve gap after 3-month delay and functional and morphometrical analyses were performed 4 months after nerve reconstruction. Our data show that the enriched chitosan conduit is as effective as the hollow chitosan conduit in promoting nerve regeneration, and its efficacy is not statistically different from the autograft, considered the “gold standard” technique for nerve reconstruction. Since hollow tubes not always lead to good results after long defects (> 20 mm), we believe that the conduit enriched with fresh muscle fibers could be a promising strategy to repair longer gaps, as muscle fibers create a favorable three-dimensional environment and release trophic factors. All procedures were approved by the Bioethical Committee of the University of Torino and by the Italian Ministry of Health (approval number: 864/2016/PR) on September 14, 2016.

Repair of peripheral nerve defects by nerve transposition using small gap bio-sleeve suture with different inner diameters at both ends.

During peripheral nerve transposition repair, if the diameter difference between transposed nerves is large or multiple distal nerves must be repaired at the same time, traditional epineurial neurorrhaphy has the problem of high tension at the suture site, which may even lead to the failure of nerve suture. We investigated whether a small gap bio-sleeve suture with different inner diameters at both ends can be used to repair a 2-mm tibial nerve defect by proximal transposition of the common peroneal nerve in rats and compared the results with the repair seen after epineurial neurorrhaphy. Three months after surgery, neurological function, nerve regeneration, and recovery of nerve innervation muscle were assessed using the tibial nerve function index, neuroelectrophysiological testing, muscle biomechanics and wet weight measurement, osmic acid staining, and hematoxylin-eosin staining. There was no obvious inflammatory reaction and neuroma formation in the tibial nerve after repair by the small gap bio-sleeve suture with different inner diameters at both ends. The conduction velocity, muscle strength, wet muscle weight, cross-sectional area of muscle fibers, and the number of new myelinated nerve fibers in the bio-sleeve suture group were similar to those in the epineurial neurorrhaphy group. Our findings indicate that small gap bio-sleeve suture with different inner diameters at both ends can achieve surgical suture between nerves of different diameters and promote regeneration and functional recovery of injured peripheral nerves.

Research progress of graphene and its derivatives in repair of peripheral nerve defect

To review the research progress of graphene and its derivatives in repair of peripheral nerve defect. The related literature of graphene and its derivatives in repair of peripheral nerve defect in recent years was extensively reviewed. It is confirmed by in vitro and in vivo experiments that graphene and its derivatives can promote cell adhesion, proliferation, differentiation and neurite growth effectively. They have good electrical conductivity, excellent mechanical properties, larger specific surface area, and other advantages when compared with traditional materials. The three-dimensional scaffold can improve the effect of nerve repair. The metabolic pathways and long-term reaction of graphene and its derivatives in the body are unclear. How to regulate their biodegradation and explain the electric coupling reaction mechanism between cells and materials also need to be further explored.

The effect of four types of artificial nerve graft structures on the repair of 10-mm rat sciatic nerve gap.

Investigating the effect of four types of artificial nerve graft (ANG) structures on rat sciatic nerve defect repair will aid future ANG designs. In this study, fibroin fibers and polylactic acid were used to prepare four ANGs with differing structures: nerve conduit with micron‐sized pores (Conduit with pore group), nerve conduit without micron‐sized pores (Conduit group), nerve scaffold comprising Conduit with pore group material plus silk fibers (Scaffold with pore group), and nerve scaffold comprising Conduit group material plus silk fibers (Scaffold group). ANGs or autologous nerves (Autologous group) were implanted into 10 mm rat sciatic nerve defects (n = 50 per group). Twenty weeks after nerve grafting, the time required to retract the surgical limb from the hot water was ranked as follows: Conduit with pore group > Scaffold with pore group > Conduit group > Scaffold group > Autologous group. The static sciatic index was ranked in descending order: Autologous group > Scaffold group > Conduit group > Scaffold with pore group > Conduit with pore group. Immunofluorescence staining identified significant differences in the distribution and number of axons, Schwann cells, and fibroblasts. These findings indicate that ANGs with micron‐sized pores had a negative impact on the repair of peripheral nerve defects, while internal microchannels were beneficial. © 2017 The Authors. Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3077–3085, 2017.

A multi-walled silk fibroin/silk sericin nerve conduit coated with poly(lactic-co-glycolic acid) sheath for peripheral nerve regeneration

The linearly oriented multi-walled silk fibroin/silk sericin (SF/SS) nerve conduits (NCs) can provide physical cues similar to native peripheral nerve fasciculi, but the mechanical properties of which are not excellent enough. In this study, NCs with a novel and bionic design with dual structures were developed. The important features of our NCs is that the internal skeleton (the multi-walled SF/SS conduits) has a bionic structure similar to the architecture of native peripheral nerve fasciculi, which is beneficial for nerve regeneration, and the outer sheath (the hollow poly(lactic-co-glycolic acid) [PLGA] conduits) could provide strong mechanical protection for the internal skeleton. The linearly oriented multi-walled SF/SS conduit was fabricated and inserted in the hollow PLGA sheath lumen and then used for the bridge across the sciatic nerve defect in rats. The outcome of the peripheral nerve repair post implantation was evaluated. The functional and morphological parameters were examined and showed that the novel PLGA-coated SF/SS NCs could promote peripheral nerve regeneration, approaching those elicited by nerve autografts that are the first candidate for repair of peripheral nerve defects. Thus, these updated NCs have potential usefulness to enhance functional recovery after repair of peripheral nerve defect.

Single injection of a novel nerve growth factor coacervate improves structural and functional regeneration after sciatic nerve injury in adult rats

The prototypical neurotrophin, nerve growth factor (NGF), plays an important role in the development and maintenance of many neurons in both the central and peripheral nervous systems, and can promote functional recovery after peripheral nerve injury in adulthood. However, repair of peripheral nerve defects is hampered by the short half-life of NGF in vivo, and treatment with either NGF alone or NGF contained in synthetic nerve conduits is inferior to the use of nerve autografts, the current gold standard. We tested the reparative ability of a single local injection of a polyvalent coacervate containing polycation-poly(ethylene argininylaspartate diglyceride; PEAD), heparin, and NGF, in adult rats following sciatic nerve crush injury, using molecular, histological and behavioral approaches. In vitro assays demonstrated that NGF was loaded into the coacervate at nearly 100% efficiency, and was protected from proteolytic degradation. In vivo, the coacervate enhanced NGF bioavailability, leading to a notable improvement in motor function (track walking analysis) after 30days. The NGF coacervate treatment was also associated with better weight gain and reduction in atrophy of the gastrocnemius muscle. Furthermore, light and electron microscopy showed that the number of myelinated axons and axon-to-fiber ratio (G-ratio) were significantly higher in NGF coacervate-treated rats compared with control groups. Expression of markers of neural tissue regeneration (MAP-2, S-100β, MBP and GAP-43), as well as proliferating Schwann cells and myelin-axon relationships (GFAP and NF200), were also increased. These observations suggest that even a single administration of NGF coacervate could have therapeutic value for peripheral nerve regeneration and functional recovery.

Preparation and characterization of injectable chitosan–hyaluronic acid hydrogels for nerve growth factor sustained release

The usage of hollow nerve conduits shows inferior recovery effect on the repair of peripheral nerve defects. In this study, a biocompatible and biodegradable pH-induced injectable chitosan–hyaluronic acid hydrogel for nerve growth factor encapsulation and sustained release was developed as the fillers in the lumen of hollow nerve conduit to reform its microenvironment for peripheral nerve regeneration. The physicochemical properties of hydrogel were characterized by gelation time, Fourier transform infrared spectroscopy, scanning electron microscopy, compressive modulus, porosity, swelling ratio, and in vitro degradation. The in vitro nerve growth factor release profiles and cell evaluation were also investigated. The results show that the structure of chitosan–hyaluronic acid hydrogel is composed of interconnected channels with a controllable pore diameter ranging from 20 to 100 µm. The hydrogel can be degraded more than 70% within 8 weeks in vitro and is available for nerve growth factor sustained release. The chitosan–hyaluronic acid/nerve growth factor hydrogel is non-toxic and suitable for adhesion and proliferation of nerve cells and capable of maintaining nerve growth factor activity. Therefore, it could be a promising intraluminal filler of nerve conduits for peripheral nerve regeneration in neural tissue engineering.

A biological tube technique for the repair of peripheral nerve defects using 'stuffed nerves'.

Presently described is research examining the "stuffed nerve" technique to repair peripheral nerve defects. Twenty-one male Wistar Albino rats were divided into 3 groups of 7, and standard 10-mm defects were created in the sciatic nerve of all subjects. Rats were treated with autogenous nerve graft (Group 1), hollow vein graft (Group 2), or vein graft stuffed with shredded nerves (Group 3). After 12 weeks, electrophysiological and histomorphological analyses were performed to evaluate axonal regeneration. Rat groups were compared in terms of latency period and peak-to-peak potential. Latency period was significantly shorter and peak-to-peak potential was significantly greater in Group 1 than in Group 2. However, latency period and peak-to-peak potential did not differ significantly between Groups 1 and 3 or between Groups 2 and 3. To evaluate axonal regeneration, number of axons, axon diameter and myelin sheath thickness was compared between groups. Results indicated that axonal regeneration was similar in Groups 1 and 3, and was better than results seen in Group 2. The stuffed nerve technique is an alternative to autogenous nerve grafting and produces similar electrophysiological and histomorphological properties.

Progress in the Research and Development of Nerve Conduits

The reconstruction after peripheral nerve damage, especially for long-segment nerve defects, remains a clinical challenge. Autologous nerve graft transplantation is an efficient method for the repair of peripheral nerve defects, but the involved complications and shortcomings have greatly limited the clinical efficacy of treatments offered to patients with nerve defects. Thus, there is an urgent need to develop new therapeutic strategies and explore alternatives to autologous nerve transplantation in clinical practice, based on the knowledge of the peripheral nerve regeneration mechanism and biological histocompatibility principles. With significant advances in the research and application of nerve conduits, they have been used to repair peripheral nerve injury for several decades. In this paper, the study background of nerve conduits, their applications in clinic, status of conduit material research and construction of tissue-engineered artificial nerves were reviewed.

Efficient bridging of 20 mm rat sciatic nerve lesions with a longitudinally micro-structured collagen scaffold

An increasing number of biomaterial nerve guides has been developed that await direct comparative testing with the ‘gold-standard’ autologous nerve graft in functional repair of peripheral nerve defects. In the present study, 20 mm rat sciatic nerve defects were bridged with either a collagen-based micro-structured nerve guide (Perimaix) or an autologous nerve graft. Axons regenerated well into the Perimaix scaffold and, the majority of these axons grew across the 20 mm defect into the distal nerve segment. In fact, both the total axon number and the number of retrogradely traced somatosensory and motor neurons extending their axons across the implant was similar between Perimaix and autologous nerve graft groups. Implantation of Schwann cell-seeded Perimaix scaffolds provided only a beneficial effect on myelination within the scaffold. Functional recovery supported by the implanted, non-seeded Perimaix scaffold was as good as that observed after the autologous nerve graft, despite the presence of thinner myelin sheaths in the Perimaix implanted nerves. These findings support the potential of the Perimaix collagen scaffold as a future off-the-shelf device for clinical applications in selected cases of traumatic peripheral nerve injury.

Nerve defect repair by differentiated adipose-derived stem cells and chondroitinase ABC-treated acellular nerves

Objective: To evaluate the effects of differentiated adipose-derived stem cells (dADSC) and chondroitinase ABC (ChABC)-treated acellular nerves (ACN) in building artificial nerves and repairing nerve defects. Methods: ADSC were isolated from the adipose tissue of Wistar rats, induced to differentiate into Schwann-like cells, and implanted into ChABC-treated ACN to repair a 15-mm sciatic nerve defect in Sprague–Dawley rats (the experimental group, group D). The control groups were an autologous nerve transplantation group (group E); ACN (group A), ChABC-treated ACN graft group (group B), and dADSC + ACN (group C). Twelve weeks after surgery, electromyography recordings, tricep surae muscle wet weight recovery rate, and axon counts were measured to evaluate the repair of peripheral nerve defects. Results: The nerve conduction velocity, compound muscle action potentials, tricep surae muscle wet weight recovery rate, and myelinated axon counts in the ChABC-ACN/dADSC group were significantly higher than in the other groups (P < 0.05), which were all lower than the autologous group (P < 0.05). Conclusions: The combination of ChABC-treated ACN and dADSC exhibited a synergistic effect in promoting nerve regeneration, and could be an alternative for effective tissue-engineered nerves.

Construction of nerve guide conduits from cellulose/soy protein composite membranes combined with Schwann cells and pyrroloquinoline quinone for the repair of peripheral nerve defect

Regeneration and functional reconstruction of peripheral nerve defects remained a significant clinical challenge. Nerve guide conduits, with seed cells or neurotrophic factors (NTFs), had been widely used to improve the repair and regeneration of injured peripheral nerve. Pyrroloquinoline quinone (PQQ) was an antioxidant that can stimulate nerve growth factors (NGFs) synthesis and accelerate the Schwann cells (SCs) proliferation and growth. In present study, three kinds of nerve guide conduits were constructed: one from cellulose/SPI hollow tube (CSC), another from CSC combined with SCs (CSSC), and the third one from CSSC combined with PQQ (CSSPC), respectively. And then they were applied to bridge and repair the sciatic nerve defect in rats, using autograft as control. Effects of different nerve guide conduits on the nerve regeneration were comparatively evaluated by general analysis, sciatic function index (SFI) and histological analysis (HE and TEM). Newly-formed regenerative nerve fibers were observed and running through the transparent nerve guide conduits 12 weeks after surgery. SFI results indicated that the reconstruction of motor function in CSSPC group was better than that in CSSC and CSC groups. HE images from the cross-sections and longitudinal-sections of the harvested regenerative nerve indicated that regenerative nerve fibers had been formed and accompanied with new blood vessels and matrix materials in the conduits. TEM images also showed that lots of fresh myelinated and non-myelinated nerve fibers had been formed. Parts of vacuolar, swollen and abnormal axons occurred in CSC and CSSC groups, while the vacuolization and swell of axons was the least serious in CSSPC group. These results indicated that CSSPC group had the most ability to repair and reconstruct the nerve structure and functions due to the comprehensive contributions from hollow CSC tube, SCs and PQQ. As a result, the CSSPC may have the potential for the applications as nerve guide conduits in the field of nerve tissue engineering.

Bone marrow-derived, neural-like cells have the characteristics of neurons to protect the peripheral nerve in microenvironment.

Effective repair of peripheral nerve defects is difficult because of the slow growth of new axonal growth. We propose that “neural-like cells” may be useful for the protection of peripheral nerve destructions. Such cells should prolong the time for the disintegration of spinal nerves, reduce lesions, and improve recovery. But the mechanism of neural-like cells in the peripheral nerve is still unclear. In this study, bone marrow-derived neural-like cells were used as seed cells. The cells were injected into the distal end of severed rabbit peripheral nerves that were no longer integrated with the central nervous system. Electromyography (EMG), immunohistochemistry, and transmission electron microscopy (TEM) were employed to analyze the development of the cells in the peripheral nerve environment. The CMAP amplitude appeared during the 5th week following surgery, at which time morphological characteristics of myelinated nerve fiber formation were observed. Bone marrow-derived neural-like cells could protect the disintegration and destruction of the injured peripheral nerve.

Repair of peripheral nerve defects with chemically extracted acellular nerve allografts loaded with neurotrophic factors-transfected bone marrow mesenchymal stem cells.

Chemically extracted acellular nerve allografts loaded with brain-derived neurotrophic factor-transfected or ciliary neurotrophic factor-transfected bone marrow mesenchymal stem cells have been shown to repair sciatic nerve injury better than chemically extracted acellular nerve allografts alone, or chemically extracted acellular nerve allografts loaded with bone marrow mesenchymal stem cells. We hypothesized that these allografts compounded with both brain-derived neurotrophic factor- and ciliary neurotrophic factor-transfected bone marrow mesenchymal stem cells may demonstrate even better effects in the repair of peripheral nerve injury. We cultured bone marrow mesenchymal stem cells expressing brain-derived neurotrophic factor and/or ciliary neurotrophic factor and used them to treat sciatic nerve injury in rats. We observed an increase in sciatic functional index, triceps wet weight recovery rate, myelin thickness, number of myelinated nerve fibers, amplitude of motor-evoked potentials and nerve conduction velocity, and a shortened latency of motor-evoked potentials when allografts loaded with both neurotrophic factors were used, compared with allografts loaded with just one factor. Thus, the combination of both brain-derived neurotrophic factor and ciliary neurotrophic factor-transfected bone marrow mesenchymal stem cells can greatly improve nerve injury.

Progress in the research and development of nerve conduits

The reconstruction after peripheral nerve damage, especially for long-segment nerve defects, remains a clinical challenge. Autologous nerve graft transplantation is an efficient method for the repair of peripheral nerve defects, but the involved complications and shortcomings have greatly limited the clinical efficacy of treatments offered to patients with nerve defects. Thus, there is an urgent need to develop new therapeutic strategies and explore alternatives to autologous nerve transplantation in clinical practice, based on the knowledge of the peripheral nerve regeneration mechanism and biological histocompatibility principles. With significant advances in the research and application of nerve conduits, they have been used to repair peripheral nerve injury for several decades. In this paper, the study background of nerve conduits, their applications in clinic, status of conduit material research and construction of tissue-engineered artificial nerves were reviewed.

Differentiation of Schwann‑like cells from human umbilical cord blood mesenchymal stem cells in vitro.

The use of artificial nerves for the repair of peripheral nerve defects is restricted by the limited sources of Schwann cells (SCs). Human mesenchymal stem cell (MSC)‑derived Schwann‑like cells are considered an alternative and desirable cell source. The aim of the present study was to establish a method of inducing directional differentiation of human umbilical cord blood (hUCB) MSCs into Schwann‑like cells. Cells isolated from hUCB were cultured in MesenCult complete medium, a specialized culture medium for MSCs, to expand hUCBMSCs. hUCBMSCs were purified by repeated changing of the medium, and they were identified by detection of the specific cell surface markers for MSCs. For differentiation of Schwann‑like cells from hUCBMSCs, the purified cells were sequentially cultured in DMEM/F12 medium with various additives. Differentiated Schwann‑like cells were identified by the detection of SC‑specific markers, including S100b, glial fibrillary acidic protein and P75, by immunocytochemisty, reverse transcription‑polymerase chain reaction and western blotting. The results demonstrated that the majority of the differentiated cells presented classical dipolar and fusiform SC morphology. Notably, a large proportion of these cells expressed the three SC markers. These results suggest that hUCBMSCs can undergo directional differentiation into Schwann‑like cells in vitro and may be an important source of SCs for the treatment of peripheral nerve defects with tissue‑engineered artificial nerves.

Incorporation of chitosan microspheres into collagen-chitosan scaffolds for the controlled release of nerve growth factor.

BackgroundArtifical nerve scaffold can be used as a promising alternative to autologous nerve grafts to enhance the repair of peripheral nerve defects. However, current nerve scaffolds lack efficient microstructure and neurotrophic support.MethodsMicrosphere–Scaffold composite was developed by incorporating chitosan microspheres loaded with nerve growth factor (NGF–CMSs) into collagen-chitosan scaffolds (CCH) with longitudinally oriented microchannels (NGF–CMSs/CCH). The morphological characterizations, in vitro release kinetics study, neurite outgrowth assay, and bioactivity assay were evaluated. After that, a 15-mm-long sciatic nerve gap in rats was bridged by the NGF–CMSs/CCH, CCH physically absorbed NGF (NGF/CCH), CCH or nerve autograft. 16 weeks after implantation, electrophysiology, fluoro-gold retrograde tracing, and nerve morphometry were performed.ResultsThe NGF–CMSs were evenly distributed throughout the longitudinally oriented microchannels of the scaffold. The NGF–CMSs/CCH was capable of sustained release of bioactive NGF within 28 days as compared with others in vitro. In vivo animal study demonstrated that the outcomes of NGF–CMSs/CCH were better than those of NGF/CCH or CCH.ConclusionOur findings suggest that incorporation of NGF–CMSs into the CCH may be a promising tool in the repair of peripheral nerve defects.

Roles of reinforced nerve conduits and low-level laser phototherapy for long gap peripheral nerve repair.

Peripheral nerve injuries are common in clinical practice because of traumas such as crushing and sectioning. Lesions of the nerve structure result in lost or diminished sensitivity and/or motor activity in the innervated territory. The degree of lesion depends on the specific nerve involved, the magnitude and type of pressure exerted, and the duration of the compression. The results of such injuries commonly include axonal degeneration and retrograde degeneration of the corresponding neurons in the spinal medulla, followed by very slow regeneration (Rochkind et al., 2001). The adverse effects on the daily activities of patients with a peripheral nerve injury are a determining factor in establishing the goals of early recovery (Rodriguez et al., 2004). The most severe form of nerve damage involves complete transection of the nerve, which results in the loss of sensory and motor function at the site of injury. Although a degree of recovery can be expected in most untreated nerve injuries, the process is slow and often incomplete. Moreover, despite considerable advances in microsurgical techniques, the functional results of peripheral nerve repair remain largely unsatisfactory. The regrowth of nerves across large gaps is particularly challenging, usually requiring a nerve graft to correctly bridge the proximal and distal nerve stumps. At present, nerve autografting is the most common treatment used to repair peripheral nerve defects. However, this recognized “gold standard” technique has a number of inherent disadvantages, such as limited availability of donor tissue (IJkema-Paassen et al., 2004), secondary deformities, potential differences in tissue structure and size (Nichols et al., 2004), and numbness at donor sites (Bini et al., 2004). Although xenografts and allografts have been proposed as alternatives to autografts, the success rate of these techniques remains poor, often resulting in immune rejection. Thus, researchers have invested considerable effort in developing synthetic nerve conduits for the repair of peripheral nerve defects.

A simple model of radial nerve injury in the rhesus monkey to evaluate peripheral nerve repair.

Current research on bone marrow stem cell transplantation and autologous or xenogenic nerve transplantation for peripheral nerve regeneration has mainly focused on the repair of peripheral nerve defects in rodents. In this study, we established a standardized experimental model of radial nerve defects in primates and evaluated the effect of repair on peripheral nerve injury. We repaired 2.5-cm lesions in the radial nerve of rhesus monkeys by transplantation of autografts, acellular allografts, or acellular allografts seeded with autologous bone marrow stem cells. Five months after surgery, regenerated nerve tissue was assessed for function, electrophysiology, and histomorphometry. Postoperative functional recovery was evaluated by the wrist-extension test. Compared with the simple autografts, the acellular allografts and allografts seeded with bone marrow stem cells facilitated remarkable recovery of the wrist-extension functions in the rhesus monkeys. This functional improvement was coupled with radial nerve distal axon growth, a higher percentage of neuron survival, increased nerve fiber density and diameter, increased myelin sheath thickness, and increased nerve conduction velocities and peak amplitudes of compound motor action potentials. Furthermore, the quality of nerve regeneration in the bone marrow stem cells-laden allografts group was comparable to that achieved with autografts. The wrist-extension test is a simple behavioral method for objective quantification of peripheral nerve regeneration.

Decellularized nerve allograft treated with chondroitinase ABC for repair of peripheral nerve defects: an experimental study

Objective To evaluate the effect of chondrotinase ABC treated decelluarilzed nerve allograft on repairing never defects in rats.Methods Nerve allografts were treated with chondrotinase ABC and decellurized using chemical detergents.The allografts were employed to repair 15 mm sciatic nerve defects in rats in the experiment group (Group A).The nerve gaps were bridged by decellularized nerve without chondrotinase ABC treatment in Group B and by autografts in Group C.Pain reaction was tested at 8 weeks and 12 weeks postoperatively.Electrophysiology,muscle wet weight of the gastrocnemius-soleus complex and the myelinated fiber density of the regenerated nerve were quantified at 12 weeks postoperatively.Results The proportion of pain reaction recovery in Group A was higher than that in Group B and Group C at 8 weeks postoperatively.At 12 weeks postoperatively,pain reaction test was positive in all the rats in Group A and Group C,whereas most of but not all the rats in Group B had a positive pain reaction.There was no significant difference in muscle weight of rats in Group A and Group C at 12 weeks.The myelinated nerve fiber density at the proximal 5 mm of the graft was higher in Group A than in Group C,whereas no significant difference was seen at the distal 10 mm of the graft.The myelinated nerve fiber density was slight lower in Group A than in Group C in the tibial nerve division 5 mm distal to the graft.Compound muscle action potential amplitude showed no differences between group A and group C.Nerve conduction velocity was slower in Group A than in Group C.The aforementioned indexes in Group B were inferior to Group A and Group C.Conclusion Chondrotinase ABC treatment facilitates axons to enter the decelluarilzed nerve allograft.This reduces target organ reinnervation time and improves functional recovery. Key words: Nerve regeneration; Chondroitin sulfate; Chondroitin sulfate proteoglycans; Ace]lular nerve