Previous studies have shown that exogenous gangliosides promote nervous system regeneration and synapse formation. In this study, 10 mm sciatic nerve segments from New Zealand rabbits were thawed from cryopreservation...Previous studies have shown that exogenous gangliosides promote nervous system regeneration and synapse formation. In this study, 10 mm sciatic nerve segments from New Zealand rabbits were thawed from cryopreservation and were used for the repair of left sciatic nerve defects through allograft bridging. Three days later, 1 mL ganglioside solution （1 g/L） was sub- cutaneously iniected into the right hind leg of rabbits. Compared with non-injected rats, muscle wet weight ratio was increased at 2-12 weeks after modeling. The quantity of myelinated fibers in regenerated sciatic nerve, myelin thickness and fiber diameter were elevated at 4-12 weeks after modeling. Sciatic nerve potential amplitude and conduction velocity were raised at 8 and 12 weeks, while conduction latencies were decreased at 12 weeks. Experimental findings indicate that ganglioside can promote the regeneration of sciatic nerve defects after repair with cryopre- served peripheral nerve allografts.展开更多
Acellular nerve ailograft preserves the basilar membrane tube and extracellular matrix, which promotes selective regeneration of neural defects via bridging. In the present study, a Sprague Dawley rat sciatic nerve wa...Acellular nerve ailograft preserves the basilar membrane tube and extracellular matrix, which promotes selective regeneration of neural defects via bridging. In the present study, a Sprague Dawley rat sciatic nerve was utilized to prepare acellular nerve allografts through the use of the chemical extraction method. Subsequently, the allograft was transplanted into a 10-mm sciatic nerve defect in Wistar rats, while autologous nerve grafts from Wistar rats served as controls. Compared with autologous nerve grafts, the acellular nerve allografts induced a greater number of degenerated nerve fibers from sural nerves, as well as a reduced misconnect rate in motor fibers, fewer acetylcholine esterase-positive sural nerves, and a greater number of carbonic anhydrase-positive sensory nerve fibers. Results demonstrated that the acellular nerve allograft exhibited significant neural selective regeneration in the process of bridging nerve defects.展开更多
The experimental design evaluated histological,mechanical,and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle,multi-directional manner to provide evidence for long-ter...The experimental design evaluated histological,mechanical,and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle,multi-directional manner to provide evidence for long-term preservation.Acellular nerve allografts from human and rats were cryopreserved in a cryoprotectant（10% fetal bovine serum,10% dimethyl sulfoxide,and 5% sucrose in RPMI1640 medium） at-80°C for 1 year,followed by thawing at 40°C or 37°C for 8 minutes.The breaking force of acellular nerve allografts was measured using a tensile test.Cell survival was determined using L-929 cell suspensions.Acellular nerve allografts were transplanted into a rat model with loss of a 15-mm segment of the left sciatic nerve.Immunohistochemistry staining was used to measure neurofilament 200 expression.Hematoxylin-eosin staining was utilized to detect relative muscle area in gastrocnemius muscle.Electron microscopy was applied to observe changes in allograft ultrastructure.There was no obvious change in morphological appearance or ultrastructure,breaking force,or cytotoxicity of human acellular nerve allografts after cryopreservation at-80°C.Moreover,there was no remarkable change in neurofilament 200 expression,myelin sheath thickness,or muscle atrophy when fresh or cryopreserved rat acellular nerve allografts were applied to repair nerve injury in rats.These results suggest that cryopreservation can greatly extend the storage duration of acellular nerve tissue allografts without concomitant alteration of the physiochemical and biological properties of the engineered tissue to be used for transplantation.展开更多
BAEKGROUND： Some experimental studies demonstrate that subcutaneous implant of allograft can significantly decrease lymphocyte infiltration and reduce immunological reaction. However, compared with autologous nerve g...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.展开更多
Recent studies suggest that acellular nerve allografts(ANA)have similar efficacy as nerve autografts in certain applications of nerve surgery.However,multiple studies also demonstrate the limitations of nerve allograf...Recent studies suggest that acellular nerve allografts(ANA)have similar efficacy as nerve autografts in certain applications of nerve surgery.However,multiple studies also demonstrate the limitations of nerve allografts,resulting in poor patient outcomes.This submission discusses a recent case series of patients who failed allograft use with subsequent histologic analyses of these allografts.Recommendations on the treatment of nerve gaps are presented,drawing from our current understanding of allograft and autograft utility in reconstruction.Factors taken into account include recipient critical nerve function,existent nerve gap,and nerve diameter.展开更多
The landscape of available technology and surgical technique has changed over the last several decades,thus leading to changes in the peripheral nerve repair surgical algorithm.Neurorrhaphy is a common procedure;howev...The landscape of available technology and surgical technique has changed over the last several decades,thus leading to changes in the peripheral nerve repair surgical algorithm.Neurorrhaphy is a common procedure;however,it is well recognized that nerve repair should be performed tensionless,thus preventing the ability to perform direct repair with a nerve gap.Historically,nerve gaps were repaired with autograft.However,autograft surgery has been associated with complications such as numbness and chronic pain,which left surgeons searching for alternatives.Nerve allografts were first utilized in the 1800s but failed due to the immune response.In the modern era,they were again utilized in the 1980s,but did not gain popularity because of the need for the use of immunosuppressants.It was evident through the 1990s that continued innovation in peripheral nerve repair was needed,as studies showed that only approximately 50% of patients with nerve gap repair achieved good or excellent outcomes.In the 2000s,the advent of an engineered nerve allograft(Avance■Nerve Graft)changed the landscape of peripheral nerve repair.Early clinical evaluation of Avance showed that adequate sensation was able to be achieved in nerve gaps up to 30 mm,providing an alternative to autografts.As engineered nerve allograft use became more conventional,studies showed 87.3% meaningful recovery in nerve gaps up to 50 mm.Furthermore,recent studies have shown that gaps between 50-70 mm have shown 69% meaningful recovery.While technology and surgical technique continue to improve,these results are promising for large nerve gap repair.展开更多
文摘Previous studies have shown that exogenous gangliosides promote nervous system regeneration and synapse formation. In this study, 10 mm sciatic nerve segments from New Zealand rabbits were thawed from cryopreservation and were used for the repair of left sciatic nerve defects through allograft bridging. Three days later, 1 mL ganglioside solution （1 g/L） was sub- cutaneously iniected into the right hind leg of rabbits. Compared with non-injected rats, muscle wet weight ratio was increased at 2-12 weeks after modeling. The quantity of myelinated fibers in regenerated sciatic nerve, myelin thickness and fiber diameter were elevated at 4-12 weeks after modeling. Sciatic nerve potential amplitude and conduction velocity were raised at 8 and 12 weeks, while conduction latencies were decreased at 12 weeks. Experimental findings indicate that ganglioside can promote the regeneration of sciatic nerve defects after repair with cryopre- served peripheral nerve allografts.
文摘Acellular nerve ailograft preserves the basilar membrane tube and extracellular matrix, which promotes selective regeneration of neural defects via bridging. In the present study, a Sprague Dawley rat sciatic nerve was utilized to prepare acellular nerve allografts through the use of the chemical extraction method. Subsequently, the allograft was transplanted into a 10-mm sciatic nerve defect in Wistar rats, while autologous nerve grafts from Wistar rats served as controls. Compared with autologous nerve grafts, the acellular nerve allografts induced a greater number of degenerated nerve fibers from sural nerves, as well as a reduced misconnect rate in motor fibers, fewer acetylcholine esterase-positive sural nerves, and a greater number of carbonic anhydrase-positive sensory nerve fibers. Results demonstrated that the acellular nerve allograft exhibited significant neural selective regeneration in the process of bridging nerve defects.
基金supported by the National Natural Science Foundation of China,No.81201546the Doctoral Start-up Program of Natural Science Foundation of Guangdong Province of China,No.2017A030310302+1 种基金the Medical Scientific Research Foundation of Guangdong Province of China,No.A2016018grants from the Science and Technology Project of Guangdong Province of China,No.2016A010103012,2013B010404019
文摘The experimental design evaluated histological,mechanical,and biological properties of allogeneic decellularized nerves after cryopreservation in a multi-angle,multi-directional manner to provide evidence for long-term preservation.Acellular nerve allografts from human and rats were cryopreserved in a cryoprotectant（10% fetal bovine serum,10% dimethyl sulfoxide,and 5% sucrose in RPMI1640 medium） at-80°C for 1 year,followed by thawing at 40°C or 37°C for 8 minutes.The breaking force of acellular nerve allografts was measured using a tensile test.Cell survival was determined using L-929 cell suspensions.Acellular nerve allografts were transplanted into a rat model with loss of a 15-mm segment of the left sciatic nerve.Immunohistochemistry staining was used to measure neurofilament 200 expression.Hematoxylin-eosin staining was utilized to detect relative muscle area in gastrocnemius muscle.Electron microscopy was applied to observe changes in allograft ultrastructure.There was no obvious change in morphological appearance or ultrastructure,breaking force,or cytotoxicity of human acellular nerve allografts after cryopreservation at-80°C.Moreover,there was no remarkable change in neurofilament 200 expression,myelin sheath thickness,or muscle atrophy when fresh or cryopreserved rat acellular nerve allografts were applied to repair nerve injury in rats.These results suggest that cryopreservation can greatly extend the storage duration of acellular nerve tissue allografts without concomitant alteration of the physiochemical and biological properties of the engineered tissue to be used for transplantation.
文摘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.
文摘Recent studies suggest that acellular nerve allografts(ANA)have similar efficacy as nerve autografts in certain applications of nerve surgery.However,multiple studies also demonstrate the limitations of nerve allografts,resulting in poor patient outcomes.This submission discusses a recent case series of patients who failed allograft use with subsequent histologic analyses of these allografts.Recommendations on the treatment of nerve gaps are presented,drawing from our current understanding of allograft and autograft utility in reconstruction.Factors taken into account include recipient critical nerve function,existent nerve gap,and nerve diameter.
文摘The landscape of available technology and surgical technique has changed over the last several decades,thus leading to changes in the peripheral nerve repair surgical algorithm.Neurorrhaphy is a common procedure;however,it is well recognized that nerve repair should be performed tensionless,thus preventing the ability to perform direct repair with a nerve gap.Historically,nerve gaps were repaired with autograft.However,autograft surgery has been associated with complications such as numbness and chronic pain,which left surgeons searching for alternatives.Nerve allografts were first utilized in the 1800s but failed due to the immune response.In the modern era,they were again utilized in the 1980s,but did not gain popularity because of the need for the use of immunosuppressants.It was evident through the 1990s that continued innovation in peripheral nerve repair was needed,as studies showed that only approximately 50% of patients with nerve gap repair achieved good or excellent outcomes.In the 2000s,the advent of an engineered nerve allograft(Avance■Nerve Graft)changed the landscape of peripheral nerve repair.Early clinical evaluation of Avance showed that adequate sensation was able to be achieved in nerve gaps up to 30 mm,providing an alternative to autografts.As engineered nerve allograft use became more conventional,studies showed 87.3% meaningful recovery in nerve gaps up to 50 mm.Furthermore,recent studies have shown that gaps between 50-70 mm have shown 69% meaningful recovery.While technology and surgical technique continue to improve,these results are promising for large nerve gap repair.
