Autologous transplantation with fewer fibers repairs large peripheral nerve defects

Peripheral nerve injury is a serious disease and its repair is challenging. A cable-style autologous graft is the gold standard for repairing long peripheral nerve defects; however, ensuring that the minimum number of transplanted nerve attains maximum therapeutic effect remains poorly understood. In this study, a rat model of common peroneal nerve defect was established by resecting a 10-mm long right common peroneal nerve. Rats receiving transplantation of the common peroneal nerve in situ were designated as the in situ graft group. Ipsilateral sural nerves（10–30 mm long） were resected to establish the one sural nerve graft group, two sural nerves cable-style nerve graft group and three sural nerves cable-style nerve graft group. Each bundle of the peroneal nerve was 10 mm long. To reduce the barrier effect due to invasion by surrounding tissue and connective-tissue overgrowth between neural stumps, small gap sleeve suture was used in both proximal and distal terminals to allow repair of the injured common peroneal nerve. At three months postoperatively, recovery of nerve function and morphology was observed using osmium tetroxide staining and functional detection. The results showed that the number of regenerated nerve fibers, common peroneal nerve function index, motor nerve conduction velocity, recovery of myodynamia, and wet weight ratios of tibialis anterior muscle were not significantly different among the one sural nerve graft group, two sural nerves cable-style nerve graft group, and three sural nerves cable-style nerve graft group. These data suggest that the repair effect achieved using one sural nerve graft with a lower number of nerve fibers is the same as that achieved using the two sural nerves cable-style nerve graft and three sural nerves cable-style nerve graft. This indicates that according to the ‘multiple amplification＇ phenomenon, one small nerve graft can provide a good therapeutic effect for a large peripheral nerve defect.

A novel artificial nerve graft for repairing longdistance sciatic nerve defects:a self-assembling peptide nanofiber scaffold-containing poly (lactic-co-glycolic acid) conduit

In this study, we developed a novel artificial nerve graft termed self-assembling peptide nanofiber scaffold （SAPNS）-containing poly（lactic-co-glycolic acid） （PLGA） conduit （SPC） and used it to bridge a 10-mm-long sciatic nerve defect in the rat. Retrograde tracing, behavioral testing and histomorphometric analyses showed that compared with the empty PLGA conduit implantation group, the SPC implantation group had a larger number of growing and extending axons, a markedly increased diameter of regenerated axons and a greater thickness of the myelin sheath in the conduit. Furthermore, there was an increase in the size of the neuromuscular junction and myofiber diameter in the target muscle. These findings suggest that the novel artificial SPC nerve graft can promote axonal regeneration and remyelination in the transected peripheral nerve and can be used for repairing peripheral nerve injury.

Biological and artificial nerve conduit for repairing peripheral nerve defect

OBJECTIVE： Recently, with the development of biological and artificial materials, the experimental and clinical studies on application of this new material-type nerve conduit for treatment of peripheral nerve defect have become the hotspot topics for professorial physicians. DATA SOURCES ： Using the terms ＂nerve conduits, peripheral nerve, nerve regeneration and nerve transplantation＂ in English, we searched Pubmed database, which was published during January 2000 to June 2006, for the literatures related to repairing peripheral nerve defect with various materials. At the same time, we also searched Chinese Technical Scientific Periodical Database at the same time period by inputting ＂ peripheral nerve defect, nerve repair, nerve regeneration and nerve graft＂ in Chinese. STUDY SELECTION ： The materials were firstly selected, and literatures about study on various materials for repairing peripheral nerve defect and their full texts were also searched. Inclusive criteria： nerve conduits related animal experiments and clinical studies. Exclusive criteria： review or repetitive studies. DATA EXTRACTION： Seventy-nine relevant literatures were collected and 30 of them met inclusive criteria and were cited. DATA SYNTHESIS ： Peripheral nerve defect, a commonly seen problem in clinic, is difficult to be solved. Autogenous nerve grafting is still the gold standard for repairing peripheral nerve defect, but because of its application limitation and possible complications, people studied nerve conduits to repair nerve defect. Nerve conduits consist of biological and artificial materials. CONCLUSION： There have been numerous reports about animal experimental and clinical studies of various nerve conduits, but nerve conduit, which is more ideal than autogenous nerve grafting, needs further clinical observation and investigation.

Effects of subcutaneous implant of peripheral nerve allograft on the regeneration of defected sciatic nerve

BAEKGROUND： Some experimental studies demonstrate that subcutaneous implant of allograft can significantly decrease lymphocyte infiltration and reduce immunological reaction. However, compared with autologous nerve grafting, what is the effect of nerve regeneration after repair？OB]EETIVE： To observe the local nervous status of the detected part of sciatic nerve repaired through subcutaneously implanting peripheral nerve allograft, and compare the effect with fresh autologous nerve grafting.DESTGN ： Contrast observation.SETTING ： Departments of Orthopaedics of Zhengzhou Fifth People＇s Hospital and First Hospital Affiliated to Chongqing Medical University.MATEREALS ： Totally 30 healthy adult Wistar male rats, with body mass of （200±20）g, were enrolled. Optical microscope （Olympus biological microscope BHS, Japan）, Electron microscope （H-600, Japan）,CM-2000 biomedical image analysis system （CM-2000,Beihang） and myoelectricity scanner （KEYPOINT, Denmark） were used in this experiment.METHODS ： This experiment was carried out in the Orthopaedic Laboratory of Chongqing Medical University between October 2000 and April 2002. ① Six rats were chosen as the donors for allogenic nerve grafting, and 15 mm sciatic nerve segment was chosen as graft. The other rats were randomly divided into two groups： allogenic nerve grafting group and autologous nerve grafting group, with 12 rats in each group. In the allogenic nerve grafting group, a skin incision was made on the posterior side of right thigh, and subcutaneous blunt dissection was performed prorsally a little, then allograft was implanted. Two weeks later, sciatic nerve was exposed at the posterior side of left thigh and cut respectively at 5 mm and another 10 mm away from pelvis. The donor nerve （with connective tissue veil） implanted subcutaneously on the right thigh was taken out. Sectioned connective tissue at the proximal end was trimmed and that at the distal end as done but reserved 10 mm in length, and inosculated antegradely at the nerve defect on the left side with 11/0 nylon line. Twelve rats in autologous nerve grafting group underwent a 10 mm sciatic nerve defect inci- sion on the right thigh and implant of fresh sciatic nerve from left thigh. The incision on the left thigh was repaired in situ. ②2,4,8 and 14 weeks after grafting, the nerve specimen of anastomosis segment was observed under optical microscope. Fourteen weeks after grafting, the ultrathin section of distal sciatic nerve was observed under transmission electron microscope. The number and size of regenerated axons at the cross section of anastomosis of proximal and distal sciatic nerve were analyzed with biomedical image analysis system. Neuroelectrophysiological change of in vivo sciatic nerve was detected with myoelectricity scanner.③ t test was used in the comparison of difference of measurement data.MAZN OUTCOME MEASURES ： ① Observation of anastomosis part of sciatic nerve under optical and electron microscopes in the two groups. ② Comparison of motor nerve conduction velocity, latent period and action potential peak as well as the number and size of cross-section of anastomosis part of proximal and distal sciatic nerve between two groups.RESULTS： ①Observation under optical microscope：Two weeks after grafting, neve axon of repaired region broke and medullary sheath denatured in the allogenic nerve grafting group and autologous nerve grafting group. At the same time, vascular engorgement and a little lymphocytes infiltration were found in the autologous nerve grafting group, but those were found worsened in the allogenic nerve grafting group. Four weeks after grafting, the intensity of the inflammatory reaction was similar between two groups, some collagen fibers at the proximal end proliferated; 8 weeks after grafting, the inflammatory reaction ended basically and the collagen fibers proliferated obviously in the two groups. ② Observation under electron microscope： Fourteen weeks after grafting, the structure of epineurium was in integrity and there were no obvious differences in perineurium and endonurium between two groups. A large number of myelinated nerve fibers and a few unmyelinated nerve fibers regenerated. The structure of myelin sheath was in integrity. ③The number and size of regenerated axons of anastomosis of proximal and distal sciatic nerve had no significant difference 14 weeks after grafting [（575.500±7.495） vs（585.700±11.172） axons/visual field ; （389.300±49.073） vs （407.600±0.283） axons/visual field;（6 423.830±119.911 ） vs （6 695.36± 84.287） μm^2/visual field = （ 5 980.110±74.572） vs（5 980.110±74.572） μm^2/visual field] （P 〉 0.05）. ④ Neuroelectrophysiological examination showed that there were no significant differences in motor nerve evoked potential latent pedod[（1.420±0.346）vs （1.237±0.250） ms] , motor nerve conduction velocity [（ 12.120±0.906 ） vs（13.020±0.599） m/s]and latent period of sciatic nerve [（0.500±0.380）vs （1.250±1.067） mV] of rats between two groups （P 〉 0.05）.CONCLUSTON： Although subcutaneous implant of peripheral nerve allograft has some inflammatory reactions, no obvious rejection is found. Repair results of two groups show that subcutaneous implant of allograft can promote nerve regeneration, which is similar to autologous nerve grafting.

Differentiation of mesenchymal stem cells into neuronal cells on fetal bovine acellular dermal matrix as a tissue engineered nerve scaffold

The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined speciifc neu-ronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuro-nal-speciifc proteins, includingβIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differen-tiation medium differentiated into a multilayered neural network-like structure with long nerve ifbers that was composed of several parallel microifbers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sec-tioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve.

Immunological rejection of acellular heterogeneous nerve transplant for bridging the sciatic nerve in rats

BACKGROUND：Artificial materials composed of acellular heterogeneous nerves can resolve donor shortage problems for the repair of peripheral nerve defects.However,it remains unclear whether artificial materials can overcome immunological rejection of heterogeneous nerve grafts and obtain similar effects as allogeneic nerve grafts.OBJECTIVE：To analyze regeneration and immunological rejection of defective sciatic nerves in rats through the use of acellular heterogeneous nerve grafts.DESIGN,TIME AND SETTING：A randomized,controlled study was performed at the Department of Anatomy,China Medical University and the Experimental Center,First Affiliated Hospital,China Medical University between January and December 2008.MATERIALS：TritonX-100 （Sigma,USA） and deoxycholate （Pierce,USA） were used.METHODS：Bilateral sciatic nerves were collected from adult rabbits and treated with TritonX-100 and sodium deoxycholate to prepare acellular sciatic nerves,which were used to bridge 1 -cm defective sciatic nerves in adult rats.MAIN OUTCOME MEASURES：The lymphocyte percentage in leukocytes was quantified following hemocyte staining.Neural regeneration and the recovery of motor end plates in the gastrocnemius muscle were observed under optical and electronic microscopy following toluidine blue staining,as well as acetylcholinesterase and succinate dehydrogenase histochemical staining.RESULTS：There was no significant difference in the lymphocyte percentage in leucocytes between transplanted and normal rats （P 〉 0.05）.At 3 months after surgery,the rat toes on the operated side were separated and the rats could walk.In addition,the footplates exhibited an escape response when acupunctured.A large number of regenerated nerve fibers were observed in the transplant group,and acetylcholinesterase-positive motor end plates were visible in fibers of the gastrocnemius muscle.CONCLUSION：Acellular heterogeneous nerve transplants for the repair of defective sciatic nerves in rats promote neural regeneration without significant immunological rejection.