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Objective To investigate the protective effect and mechanism of acellular nerve allografts(ANA)combined with electroacupuncture on spinal ganglia in rats with sciatic nerve injury(SNI).Methods Totally 50 male adult SD rats were randomly selected for this experiment.Ten rats were prepared for the ANA.Forty male SD rats were randomly divided into normal group,model group,ANA group and combinational group,with 10 rats in each group.The SNI model was established by cutting off the nerves 10 mm at the 5 mm on the inferior border of piriformis after separating the right sciatic nerves.The rats in the ANA group were bridged with ANA to the two broken ends of injured nerves.The rats in the combinational group were treated with electroacupuncture 2 days after ANA bridging,Huantiao(GB30)and Yanglingquan(GB34)were performed as the acupuncture points,each electroacupuncture lasted 15 minutes and 7 days as a course of treatment,4 courses in all.Sciatic nerve conduction velocity was measured by electrophysiology to evaluate the regeneration of damaged axons.Morphology of spinal ganglia was observed by Nissl staining.The expression of nerve growth factor(NGF)and brain-derived neurotrophic factor(BDNF)were detected by Western blotting and immunofluorescent staining.Results Compared with the normal group,the sciatic nerve conduction velocity in model group decreased significantly(P<0.01),Nissl bodies in neurons of spinal ganglia were swollen and dissolved,with incomplete structure and the number decreased dramatically(P<0.01),while the level of NGF and BDNF also decreased significantly(P<0.01).Compared with the model group,the sciatic nerve conduction velocity in ANA and combinational groups strongly increased(P<0.01),the damage of Nissl bodies in neurons of spinal ganglia reduced and the number obviously increased(P<0.01),the level of NGF and BDNF increased considerably(P<0.01).Compared with the ANA group,the sciatic nerve conduction velocity in combinational group increased significantly(P<0.01),the morphology of Nissl bodies in neurons of spinal ganglia were more regular and the number increased(P<0.01),moreover,the level of NGF also increased significantly(P<0.01).Conclusion ANA combined with electroacupuncture can enhance the sciatic nerve conduction velocity,improve the morphology of neurons in spinal ganglia and play a protective effect on spinal ganglia.The mechanism can be related to the higher expression of NGF and BDNF proteins,especially the expression of NGF protein.
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Objective:To observe clinical outcomes of the repair of traumatic nerve defects in the proximal upper extremities by human acellular nerve allograft(hANG).Methods:Nerve defects in the upper extremities in 8 patients were repaired by hANG from March 2017 to January 2019. The patients were 6 males and 2 females with mean age of 35.4 (21-53) years old. The nerve defects were 2 radial nerve in distal upper arm, 4 median nerve in forearm, 1 interosseous dorsal nerve and 1 ulnar nerve in forearm. All injuries were acute nerve injury. Two patients had combined injury of upper arm muscle, 4 of forearm muscle and 1 of brachial artery defect. All wound were moderate to severe contaminated. The length of nerve defects was 30-60 (mean 45) mm. The surgical procedures were fixation of fracture, repair of the muscle and discovery of the broken ends of nerve and to repair with hANG. The postoperative follow-up period ranged 18 to 40 (mean 30.6) months to observe the local response of recovery. The efficacy was evaluated by the Upper Extremity Function Evaluation Standard set up by Hand Surgery Branch of Chinese Medical Association and Grading Standard of Muscle Strength.Results:No graft rejection was observed in all cases. Primarily healing was in 5 patients. Delayed healing in 2 patients and free skin grafting was performed. Local flap transfer was performed to repair the wound in 1 patient who developed a skin necrosis 10 days after surgery. Two patients with median nerve defects had nerve function restored well. The strength of finger grip and thumb opposition muscle restored to grade IV and the sensory function had restored S 3+. The interosseous dorsal nerve in 1 patient restored well. The strength of extensor digitorum tendon had restored to grade IV. Based on the evaluation criteria for the upper extremity issued by the Hand Surgery of the Chinese Medical Association, 3 patients was rated in excellent for function recovery, 1 in fair and 4 in poor. Conclusion:After throughout debridement, hANG can be applied in the repair of traumatic nerve defect in the proximal upper extremity in an emergency surgery and it can partially restore the nerve function.
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BACKGROUND: It is seldom reported that the effect of chemical acellular nerve allograft combined with bone marrow mesenchymal stem cell transplantation on sciatic nerve injury is evaluated by tensile mechanical properties. OBJECTIVE: To determine the effect of chemical acellular nerve allograft combined with bone marrow mesenchymal stem cell transplantation in the treatment of sciatic nerve injury. METHODS: Forty-five Sprague-Dawley rats were randomly divided into autologous nerve transplantation group, chemical acellular nerve allograft group, and chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation group (n=15 per group). Models of 10-mm sciatic nerve injury were prepared. The injured sciatic nerves were repaired using autologous nerve, chemical acellular nerve allograft and chemical acellular nerve allograft and bone marrow mesenchymal stem cells. At 20 weeks after surgery, electrophysiological measurement and stretching experiments were carried out. RESULTS AND CONCLUSION: (1) The wave amplitude and motion conduction velocity were larger in the chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation group and autologous nerve transplantation group than in the chemical acellular nerve allograft group (P < 0.05). (2) The tensile elastic limit strain, elastic limit stress, maximum strain and maximum stress were larger in the chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation group and autologous nerve transplantation group than in the chemical acellular nerve allograft group (P < 0.05). (3) These results suggest that chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation has obvious effect on repairing sciatic nerve injury.
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La reconstrucción de nervios periféricos con aloinjertos nerviosos acelulares humanos en neurocirugía ha sido bastante estudiada estableciendo su predictibilidad y éxito en intervenciones principalmente en los nervios digitales de las manos. En cirugía maxilofacial existe una creciente investigación para poder restaurar el nervio alveolar inferior en cirugías de resección mandibular en donde la extirpación de esta estructura nerviosa es inevitable. El objetivo de esta publicación es mostrar un reporte de un caso en donde se realizó la reconstrucción del nervio alveolar inferior con aloinjerto de nervio acelular humano (Avance® Nerve Graft, Axogen) con microcirugía para poder proveer de sensibilidad a la región de la cara afectada en un paciente reconstruido con un injerto de fíbula microvascularizada posterior a una hemimandibulectomía por ameloblastoma plexiforme.
The reconstruction of peripheral nerves with allografts of human acellular nerves in neurosurgery is well studied, establishing its predictability and success in interventions mainly in the digital nerves of the hands. In maxillofacial surgery there is a growing investigation to be able to restore the inferior alveolar nerve in mandibular resection surgeries where the removal of this nervous structure is inevitable. The objective of this publication is to show a case report in which the reconstruction of the inferior alveolar nerve was performed with human acellular nerve allograft (Avance® Nerve Graft, Axogen) with microsurgery in order to provide sensitivity to the region of the affected face in a reconstructed patient with a microvascularized bone fibula graft after hemimandibulectomy due to plexiform ameloblastoma.
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Humans , Male , Adolescent , Peripheral Nerves/transplantation , Neurosurgical Procedures/methods , Mandibular Nerve/surgery , AllograftsABSTRACT
Objective To discuss the effect of bone marrow mesenchymal stem cell (BMSC)as the seed cell transplantation of tissue-engineered artificial nerve in the treatment of peripheral nerve injury. Methods BMSC was obtained from the bone marrow of adult rat through isolation and culture and combined with acellular nerve scaffold to construct ‘tissue-engineered artificial nerve’.After transplantation,rats were divided into two groups,the BMSC +acellular nerve conduit group(BMSC treatment group)and the empty cell conduit group(negative control group)with 5 rats in each group.Sciatic functional index (SFI)of the affected side of rats was compared between two groups at 2 weeks,4 weeks and 8 weeks after the surgery.Moreover,the sciatic conduction,recovery rate of tricipital muscle wet weight and other repair effects of the affected side were compared between two groups at 8 weeks after the surgery.Results The indicators of BMSC treatment group, including SFI assessed at 2 weeks,4 weeks and 8 weeks after the surgery as well as the sciatic conduction and recovery rate of tricipital muscle wet weight assessed at 8 weeks after the surgery,were better than those of the negative control group(all in P <0.05).Conclusions BMSC combined with tissue-engineered artificial nerve of acellular nerve scaffold can effectively promote nerve regeneration and function recovery.
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Objective To investigate clinical efficacy of tissue-engineered artificial nerve grafts constructed by acellular nerve grafts combined with adult rat Schwann cell (SC)in the treatment of peripheral nerve injury.Methods SCs were isolated and cultured from the distal nerves of adult Sprague Dawley (SD) rats with 1-week Wallerian degeneration and then combined with acellular nerve grafts to construct tissue-engineered artificial nerve.All rats were divided into acellular nerve graft containing SCs (SC group)and nerve graft containing no cells groups (control group),five animals in each group.At 2-,4-and 8-week after surgery,sciatic function index (SFI)of the affected side was compared between two groups.At postoperative 8 weeks,nerve conduction of sciatic nerve of the injured side,recovery rate of triceps surae wet weight and other relevant parameters were equally compared between two groups.Results In the SC group,SFI of the affected side at 2-,4-and 8-week after surgery,nerve conduction of sciatic nerve at the injured side and recovery rate of triceps surae wet weight at postoperative 8 weeks were significantly better compared with those in the control group (all in P <0.05).Conclusions Combined use of adult rat SCs and acellular nerve grafts effectively repairs peripheral nerve defects and accelerates functional recovery of injured nerves.
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Objective To observe the way of vascularization of acellular nerves and evaluate the enhanced vascularization of using COMP-Ang-1 into acellular nerve on bridging sciatic nerve gaps by radiography.Methods From March,2013 to June,2014,acellular nerves were harvested by chemical extraction.Thirty-six female rats weighing 200-250 g were randomly divided into 2 groups:18 animals with 1 cm long sciatic nerve lesions were repaired by nerve grafting (control group),18 animals with 1 cm long sciatic nerve lesions were repaired by nerve grafting and COMP-Ang-1 were administrated after surgery.Grafts were harvested after perfusion of lead oxide (carotid artery) on day 7,day 14 and day 21 postoperatively.Radiography was performed to capture the two dimensional image.The rules of vascularization of acellular nerve and the enhanced effects of COMP-Ang-1 on vascularization were evaluated.Results The density of vessels in COMP-Ang-1 group were higher than control group after 7 days (2701.60 ± 318.93 vs.925.40 ± 106.22,P =0.030),14 days (3309.21 ± 381.31 vs.2832.70 ± 189.23,P =0.210) and 21 days (4787.33 ± 251.09 vs.3469.36 ± 232.10,P =0.030) postoperatively; the area of vessels in COMP-Ang-1 group were higher than control group after 7 days (9231.03 ± 581.91 μm2 vs.4839.01 ± 101.01 μm2,P =0.043) and 14 days (15561.13 ± 697.73 vs.6811.07 ± 250.05,P =0.049) postoperatively.Conclusion COMP-Ang-1 can enhance the vascularization of acellular nerves fairly.
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Objective To explore the safety and clinical effect of the human acellular nerve allograft (hANG) for repairing peripheral nerve defects.Methods During November,2009 to October,2010,6 patients with 3 digital nerve defects and 3 radial nerve defects were repaired with hANG.During postoperation period,safety was evaluated by local wound response and laboratory testing,while the efficacy was evaluated by British Medical Research Council sensory function assessment standards,static 2-point discrimination (2PD) and Semmes-Weinstein monofilament testing.Results Three patients with 6 digital nerve defects received hANG transplant.The length of nerve graft was 20-50 mm(mean 30.8 mm).After followed up for 31-40 months,the excellent rate of 2PD was 66.7%.Two of 3 patients rahabilited as well as the normal.Three patients with radial nerve defects,whose length of nerve graft was 35-60 mm(mean 48.3 mm).The strength of extensor carpiradialis longus muscle had restored Ⅲ in 1 case,and other 2 cases had no restoration.Conclusion hANG is safe and effective for repairing peripheral nerve defects,especially for digital nerve defects.
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Objective: To observe the biocompatibility of acellular nerve scaffold (ANS) via three sterilization methods, to provide experimental data for tissue engineering industrialization. Methods: Pig sciatic nerves were cut and treated using the NaOH maceration method. ANSs were sterilized by ethylene oxide, ~(60)Co-irradiation and peracetic acid. Evaluated the biocompatibility by MTT, cellular compatibility test, collagenase susceptibility test in vitro and local implantation test. Results: ANS retained the integrity of structure and major components of the basement membrane. The result of MTT test showed that the ANSs via different sterilization methods had statistical differences. There were no overall significant differences in Collagenase susceptibility test. Scanning electron microscope results showed the skin fibroblasts could attach, proliferate and grow well on the surface and holes of ANS with sterilization of PAA and Co~(60),a small quantity of cells adhered on ANS with sterilization of ETO. Tests for local effects after implantation show that different sterilization methods don't effect the ability of ANS to resist the enzyme degradation. In ETO group, rats showed an acute inflammatory response followed by chronic inflammation. In PAA and ~(60)Co group rats showed an acute inflammatory response that diminished such that the graft ultimately became indistinguishable from native tissue, observations that were consistent with graft acceptance. Conclusion: Peracetic acid sterilization offers a convenient alternative protocol for ANS processing. ANS sterilized with PAA shows good compatibility and biologic safety. It is an ideal sterilization method for ANS.
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Objective To explore the feasibility of using human umbilical cord derived mesenchymal stem cells as seed cells to repair sciatic nerve defects of rats by tissue engineering methods. Methods Mesenchymal stem cells from human umbilical cord were cultured and induced into neuron-liked cells,which were co-cultured with acellular basal lamina tube to construct tissue engineering nerve;models of sciatic nerve defects 10 mm in length were set up with thirty healthy adult SD rats and were divided randomly into 3 groups:tissue engineering nerve group (group A, compound of human umbilical cord derived mesenchymal stem cells and acellular basal lamina tube), pure acellular basal lamina tube group (group B), and autogenous nerve bridging group (group C). Evaluation of electrophysiological and histological results was carried out 10 weeks after operation. Results The engineering nerve group had good result in nerve regeneration which was close to the effect of autogenous nerve transfer group (group A), and much better than the effect of pure acellular basal lamina tube group. Conclusion Engineering nerves from human umbilical cord derived mesenchymal stem cells can effectively repair 10 mm defects of sciatic nerve.
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[Objective]The purpose of this paper is to demonstrate whether the nerve length could affect the quality of acellular nerve and investigate the properly repairable distance of nerve defects with acellular nerve allografts.[Method]Fresh sciatic nerves were obtained from adult dogs and divided into 12 cm long segments.The nerve segments were decellularized via an improved chemical decelluarization treatment as following: Nerve segments were rinsed with cold sterile Ringer's solution and submergedin 5% Triton-100 solution 12h,and then soaked the nerve segments into 5% sodium deoxycholate for 12h.The treated nerve segments were washed in distilled water for 3h.This procedures were repeated once again.In vitro,the degrees of decellularization,demyelination and integrity of nerve fiber tubal of chemically extracted acellular nerves were observed with microscope and assessed by a score system.In vivo,the sciatic nerve of dogs on the right was exposed.In 8 cm grafted group(n=6),a 7 cm segment of sciatic nerve was removed from the midthigh level.In 10 cm grafted group(n=6),a 9 cm segment of sciatic nerve was removed at the same level.The gaps were bridged with acellular nerve allografts by 8 cm and 10 cm long segments respectively.The follow-up period was 12 month postoperatively.Motor functional recovery of the right hind following allografting was examined by neurobehavioral,electrophysiological,histological and immunohistochemical assessment.[Result]There was no difference on the degrees of decellularization,demyelination and integrity of nerve fiber tubal among every fraction of the acellular nerve from the two ends to the central portion.In 8 cm grafted group,all survival dogs(n=5) were held upright with the affected hindlimb extended so that the body's weight was supported by the distal metatarsus and toes.In 10 cm grafted group,animals were failed to held upright with the affected hindlimb.Electrophysiological studies showed that elctromyographic activity was observed in both groups.After 12 month the conduction velocity was 32.1+5.1 m/s in 8 cm grafted animals and 18.3+6.0m/s in 10 cm grafted group.In normal animals, the conduction velocity was 106.6+16.4 m/s.The conduction velocity in 10 cm grafted group was lower than 8 cm grafted(P
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0.05).There were better effect of removal of myelin(P2
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[Objective]To evaluate the effect of post-operative rehabilitative nursing of patients repaired with acellular nerve allograft.[Method]From April 2003 to April 2006,39 inpatients with peripheral nerve defect were subjected to receive acellular nerve allograft in order to repair nerve defect.The patients were rehabilitated with special nursing after being operated and discharged.Among of them,21 patients were followed up over 6 months,the effect of treatment was analyzed.[Result]Among 21 patients,16 people had excellent and good effect of treatment and the efficient rate was 71.4%.[Conclusion]Post-operative rehabili tative nursing is important and effective for rehabilitation patients of peripheral nerve injuries repaired with acellular nerve allograft.
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[Objective]To compare two preparing procedures for larger chemically acellular nerve allografts (CANA). [Methods]The sciatic nerves of pigs were exposed by a muscle-splitting incision and were isolated free of the underlying fascia. The 60-mm-long segments of the nerve were obtained. They were treated according to the following decellularization processes.In group I,the nerve segments were treated with 7% Triton-100 solution and 7% sodium deoxycholate for two times.In group II,another protocol was created with the detergents Triton X-200,sulfobetaine-16,and sulfobetaine-10 for two times. The degrees of decellularization,activity of laminin,degrees of demyelination,and integrity of the nerve fiber tube were observed under microscope and were assessed by a scoring system.[Results]In both experimental groups the activity of laminin was present and the degrees of decellularization were complete. As for the demyelization of the nerve segments,the myelin sheath in Group II was partially preserved,but it completely disappeared in Group Ⅰ. The structure of the nerve fiber tube in Groups Ⅰ and Ⅱ were not as integral as that in the normal group.[Conclusion]It may be a better method for the larger CANA,to be treated with TritonX-100 and sodium deoxycholate during the decellularization procedure.
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Objective To investigate the activity of gastrocnemius muscle after acellular nerve transplantation with microencapsulated peripheral nerve tissue to repair sciatic nerve gap in dogs. Methods Twelve dogs were selected and divided randomly into four groups with three in each. Both sciatic nerves of the dogs in group A were cut and chemically extracted to make acellular nerves. Acellular nerves were transplanted into the dogs of groups B and C to bridge 30 mm sciatic nerve gaps, and additionally microencapsulated peripheral nerve were added in group B. Autograft was used for repairing the defects in group D. The motor function of operated limbs were observed regularly after operation. Six months after operation, motor conduction velocity of the specimens of sciatic grafting nerve segment were observed, and the gastrocnemius muscle of both sides were obtained and performed succinic dehydrogenase (SDH), synaptophysin (SYP) and motor end plate histochemical staining, and sciatic nerve 1 cm from distal end stoma were obtained and underwent Masson, Luxol fast blue and NF200 staining. Results All the dogs in groups B, C and D were involved in the results analysis without loss. Image analysis indicated the optical density and the cross section area of gastrocnemius muscle, area and optical density of synaptophysin, area and optical density of motor end plate in group B showed statistically significant difference compared with those in group C, but no significant difference with those in group D. The results of motor function recovery of operated-limb, electrophysiological examination and morphologic observations of sciatic nerve were all consistent with the detections of gastrocnemius muscle. Conclusion The gastrocnemius muscle may be again innerved by nerve, and the muscle atrophy is significantly alleviated after acellular nerve transplantation with microencapsulated peripheral nerve tissue for repairing sciatic nerve gap. The regeneration of nerve was better after the combined transplantation than only acellular nerve transplantation, so the combination of acellular nerve and microencapsulated peripheral nerve tissue may be a useful substitute of autograft for nerve repair.
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0.05). Conclusion The nerve bridge compounded of bFGF heparin, acellular nerve basal lamina tube and Schwann cells can facilitate nerve regeneration. [Key words] Peripheral nerve; Tissue engineer; Acellular nerve basal lamina tube; Basic fibroblast growth factor(bFGF); Schwann cells
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Objective To observe the repairing effect of the acellular nerve allografts on the sciatic nerve gap of rat. Methods The acellular nerve allografts,treated by hypotonic-chemical detergent,were put on the 10 mm gap of the sciatic nerve in the rat.The action potential of the regenerated nerves was determined by the electrophysiologic method 13 weeks after operation.The morphology of the regenerated nerves was observed under light microscope and electron microscope,and the results were analyzed statistically. Results No inflammation and rejected reaction were found in the period of 13 weeks after operation in the operated and control groups.There was no significant difference in number of the regenerated nerve fibers,diameter of the axons,and the thickness of the regenerated myelinated nerve between the experimental group and control group.Conclusion The present results indicated that the acellular nerve allografts had good biocompatibility for the host rat in vivo and might as a bridge promote the regeneration of the injured sciatic nerve.;