Strain and stress variations in the human amniotic membrane and fresh corpse autologous sciatic nerve anastomosis in a model of sciatic nerve injury

A 10-mm long sciatic nerve injury model was established in fresh normal Chinese patient cadavers. Amniotic membrane was harvested from healthy maternal placentas and was prepared into multilayered,coiled,tubular specimens.Sciatic nerve injury models were respectively anastomosed using the autologous cadaveric sciatic nerve and human amniotic membrane.Tensile test results showed that maximal loading,maximal displacement,maximal stress,and maximal strain of sciatic nerve injury models anastomosed with human amniotic membrane were greater than those in the autologous nerve anastomosis group.The strain-stress curves of the human amniotic membrane and sciatic nerves indicated exponential change at the first phase,which became elastic deformation curves at the second and third phases,and displayed plastic deformation curves at the fourth phase,at which point the specimens lost their bearing capacity.Experimental findings suggested that human amniotic membranes and autologous sciatic nerves exhibit similar stress-strain curves, good elastic properties,and certain strain and stress capabilities in anastomosis of the injured sciatic nerve.

Changes in proteins related to early nerve repair in a rat model of sciatic nerve injury

Peripheral nerves have a limited capacity for self-repair and those that are severely damaged or have significant defects are challenging to repair. Investigating the pathophysiology of peripheral nerve repair is important for the clinical treatment of peripheral nerve repair and regeneration. In this study, rat models of right sciatic nerve injury were established by a clamping method. Protein chip assay was performed to quantify the levels of neurotrophic, inflammation-related, chemotaxis-related and cell generation-related factors in the sciatic nerve within 7 days after injury. The results revealed that the expression levels of neurotrophic factors(ciliary neurotrophic factor) and inflammationrelated factors(intercellular cell adhesion molecule-1, interferon γ, interleukin-1α, interleukin-2, interleukin-4, interleukin-6, monocyte chemoattractant protein-1, prolactin R, receptor of advanced glycation end products and tumor necrosis factor-α), chemotaxis-related factors(cytokine-induced neutrophil chemoattractant-1, L-selectin and platelet-derived growth factor-AA) and cell generation-related factors(granulocyte-macrophage colony-stimulating factor) followed different trajectories. These findings will help clarify the pathophysiology of sciatic nerve injury repair and develop clinical treatments of peripheral nerve injury. This study was approved by the Ethics Committee of Peking University People's Hospital of China(approval No. 2015-50) on December 9, 2015.

Expression and effect of sodium-potassium-chloride cotransporter on dorsal root ganglion neurons in a rat model of chronic constriction injury

Sodium-potassium-chloride cotransporter 1 (NKCC1) and potassium-chloride cotransporter 2 (KCC2) are associated with the transmission of peripheral pain.We investigated whether the increase of NKCC1 and KCC2 is associated with peripheral pain transmission in dorsal root ganglion neurons.To this aim,rats with persistent hyperalgesia were randomly divided into four groups.Rats in the control group received no treatment,and the rat sciatic nerve was only exposed in the sham group.Rats in the chronic constriction injury group were established into chronic constriction injury models by ligating sciatic nerve and rats were given bumetanide,an inhibitor of NKCC1,based on chronic constriction injury modeling in the chronic constriction injury + bumetanide group.In the experiment measuring thermal withdrawal latency,bumetanide (15 mg/kg) was intravenously administered.In the patch clamp experiment,bumetanide (10 μg/μL) and acutely isolated dorsal root ganglion neurons (on day 14) were incubated for 1 hour,or bumetanide (5 μg/μL) was intrathecally injected.The Hargreaves test was conducted to detect changes in thermal hyperalgesia in rats.We found that the thermal withdrawal latency of rats was significantly decreased on days 7,14,and 21 after model establishment.After intravenous injection of bumetanide,the reduction in thermal retraction latency caused by model establishment was significantly inhibited.Immunohistochemistry and western blot assay results revealed that the immune response and protein expression of NKCC1 in dorsal root ganglion neurons of the chronic constriction injury group increased significantly on days 7,14,and 21 after model establishment.No immune response or protein expression of KCC2 was observed in dorsal root ganglion neurons before and after model establishment.The Cl^– (chloride ion) fluorescent probe technique was used to evaluate the change of Cl^– concentration in dorsal root ganglion neurons of chronic constriction injury model rats.We found that the relative optical density of N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (a Cl^– fluorescent probe whose fluorescence Cenintensity decreases as Cl– concentration increases) in the dorsal root ganglion neurons of the chronic constriction injury group was significantly decreased on days 7 and 14 after model establishment.The whole-cell patch clamp technique revealed that the resting potential and action potential frequency of dorsal root ganglion neurons increased,and the threshold and rheobase of action potentials decreased in the chronic constriction injury group on day 14 after model establishment.After bumetanide administration,the above indicators were significantly suppressed.These results confirm that CCI can induce abnormal overexpression of NKCC1,thereby increasing the Cl^– concentration in dorsal root ganglion neurons;this then enhances the excitability of dorsal root ganglion neurons and ultimately promotes hyperalgesia and allodynia.In addition,bumetanide can achieve analgesic effects.All experiments were approved by the Institutional Ethics Review Board at the First Affiliated Hospital,College of Medicine,Shihezi University,China on February 22,2017 (approval No.A2017-169-01).

Autologous nerve graft repair of different degrees of sciatic nerve defect:stress and displacement at the anastomosis in a three-dimensional finite element simulation model

In the repair of peripheral nerve injury using autologous or synthetic nerve grafting, the mag- nitude of tensile forces at the anastomosis affects its response to physiological stress and the ultimate success of the treatment. One-dimensional stretching is commonly used to measure changes in tensile stress and strain; however, the accuracy of this simple method is limited. There- fore, in the present study, we established three-dimensional finite element models of sciatic nerve defects repaired by autologous nerve grafts. Using PRO E 5.0 finite element simulation software, we calculated the maximum stress and displacement of an anastomosis under a 5 N load in 10-, 20-, 30-, 40-mm long autologous nerve grafts. We found that maximum displacement increased with graft length, consistent with specimen force. These findings indicate that three-dimensional finite element simulation is a feasible method for analyzing stress and displacement at the anas- tomosis after autologous nerve grafting.

The longitudinal epineural incision and complete nerve transection method for modeling sciatic nerve injury

Injury severity, operative technique and nerve regeneration are important factors to consider when constructing a model of peripheral nerve injury. Here, we present a novel peripheral nerve injury model and compare it with the complete sciatic nerve transection method. In the experimental group, under a microscope, a 3-mm longitudinal incision was made in the epineurium of the sciatic nerve to reveal the nerve fibers, which were then transected. The small, longitudinal incision in the epineurium was then sutured closed, requiring no stump anastomosis. In the control group, the sciatic nerve was completely transected, and the epineurium was repaired by anastomosis. At 2 and 4 weeks after surgery, Wallerian degeneration was observed in both groups. In the experimental group, at 8 and 12 weeks after surgery, distinct medullary nerve fibers and axons were observed in the injured sciatic nerve. Regular, dense myelin sheaths were visible, as well as some scarring. By 12 weeks, the myelin sheaths were normal and intact, and a tight lamellar structure was observed. Functionally, limb movement and nerve conduction recovered in the injured region between 4 and 12 weeks. The present results demonstrate that longitudinal epineural incision with nerve transection can stably replicate a model of Sunderland grade IV peripheral nerve injury. Compared with the complete sciatic nerve transection model, our method reduced the difficulties of micromanipulation and surgery time, and resulted in good stump restoration, nerve regeneration, and functional recovery.

Mechanism of persistent hyperalgesia in neuropathic pain caused by chronic constriction injury

Transmembrane member 16 A(TMEM16 A) is involved in many physiological functions, such as epithelial secretion, sensory conduction, nociception, control of neuronal excitability, and regulation of smooth muscle contraction, and may be important in peripheral pain transmission. To explore the role of TMEM16 A in the persistent hyperalgesia that results from chronic constriction injury-induced neuropathic pain, a rat model of the condition was established by ligating the left sciatic nerve. A TMEM16 A selective antagonist(10 μg T16 Ainh-A01) was intrathecally injected at L5–6. For measurement of thermal hyperalgesia, the drug was administered once at 14 days and thermal withdrawal latency was recorded with an analgesia meter. For measurement of other indexes, the drug was administered at 12 days,once every 6 hours, totally five times. The measurements were performed at 14 days. Western blot assay was conducted to analyze TMEM16 A expression in the L4–6 dorsal root ganglion. Immunofluorescence staining was used to detect the immunoreactivity of TMEM16 A in the L4–6 dorsal root ganglion on the injured side. Patch clamp was used to detect electrophysiological changes in the neurons in the L4–6 dorsal root ganglion. Our results demonstrated that thermal withdrawal latency was shortened in the model rats compared with control rats.Additionally, TMEM16 A expression and the number of TMEM16 A positive cells in the L4–6 dorsal root ganglion were higher in the model rats, which induced excitation of the neurons in the L4–6 dorsal root ganglion. These findings were inhibited by T16 Ainh-A01 and confirm that TMEM16 A plays a key role in persistent chronic constriction injury-induced hyperalgesia. Thus, inhibiting TMEM16 A might be a novel pharmacological intervention for neuropathic pain. All experimental protocols were approved by the Animal Ethics Committee at the First Affiliated Hospital of Shihezi University School of Medicine, China(approval No. A2017-170-01) on February 27, 2017.

Microencapsulation improves inhibitory effects of transplanted olfactory ensheathing cells on pain after sciatic nerve injury

Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells（OECs） remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4–5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4–5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.

Guinea pigs as an animal model for sciatic nerve injury

The overwhelming use of rat models in nerve regeneration studies is likely to induce skewness in treatment outcomes.To address the problem,this study was conducted in 8 adult guinea pigs of either sex to investigate the suitability of guinea pig as an alternative model for nerve regeneration studies.A crush injury was inflicted to the sciatic nerve of the left limb,which led to significant decrease in the pain perception and neurorecovery up to the 4th weak.Lengthening of foot print and shortening of toe spread were observed in the paw after nerve injury.A 3.49 ± 0.35 fold increase in expression of neuropilin 1（NRP1） gene and 2.09 ± 0.51 fold increase in neuropilin 2（NRP2） gene were recorded 1 week after nerve injury as compared to the normal nerve.Ratios of gastrocnemius muscle weight and volume of the experimental limb to control limb showed more than 50% decrease on the 30 th day.Histopathologically,vacuolated appearance of the nerve was observed with presence of degenerated myelin debris in digestion chambers.Gastrocnemius muscle also showed degenerative changes.Scanning electron microscopy revealed loose and rough arrangement of connective tissue fibrils and presence of large spherical globules in crushed sciatic nerve.The findings suggest that guinea pigs could be used as an alternative animal model for nerve regeneration studies and might be preferred over rats due to their cooperative nature while recording different parameters.

Viscoelasticity of repaired sciatic nerve by poly(lactic-co-glycolic acid) tubes

Medical-grade synthetic poly（lactic-co-glycolic acid） polymer can be used as a biomaterial for nerve repair because of its good biocompatibility, biodegradability and adjustable degradation rate. The stress relaxation and creep properties of peripheral nerve can be greatly improved by repair with poly（lactic-co-glycolic acid） tubes. ＂Fen sciatic nerve specimens were harvested from fresh corpses within 24 hours of death, and were prepared into sciatic nerve injury models by creating a 10 mm defect in each specimen. Defects were repaired by anastomosis with nerve autografts and poly（lactic-co-glycolic acid） tubes. Stress relaxation and creep testing showed that at 7 200 seconds the sciatic nerve anastomosed by poly（lactic-co-glycolic acid） tubes exhibited a greater decrease in stress and increase in strain than those anastomosed by nerve autografts. These findings suggest that poly（lactic-co-glycolic acid） exhibits good viscoelasticity to meet the biomechanical require- ments for a biomaterial used to repair sciatic nerve injury.

Poly(lactic-co-glycolic acid) conduit for repair of injured sciatic nerve A mechanical analysis

Tensile stress and tensile strain directly affect the quality of nerve regeneration after bridging nerve defects by poly（lactic-co-glycolic acid） conduit transplantation and autogenous nerve grafting for sciatic nerve injury. This study collected the sciatic nerve from the gluteus maximus muscle from fresh human cadaver, and established 10-mm-long sciatic nerve injury models by removing the ischium, following which poly（lactic-co-glycolic acid） conduits or autogenous nerve grafts were transplanted. Scanning electron microscopy revealed that the axon and myelin sheath were torn, and the vessels of basilar membrane were obstructed in the poly（lactic-co-glycolic acid） conduit-repaired sciatic nerve following tensile testing. There were no significant differences in tensile tests with autogenous nerve graft-repaired sciatic nerve. Following poly（lactic-co-glycolic acid） conduit transplantation for sciatic nerve repair, tensile test results suggest that maximum tensile load, maximum stress, elastic limit load and elastic limit stress increased compared with autogenous nerve grafts, but elastic limit strain and maximum strain decreased. Moreover, the tendencies of stress-strain curves of sciatic nerves were similar after transplantation of poly（lactic-co-glycolic acid） conduits or autogenous nerve grafts. Results showed that after transplantation in vitro for sciatic nerve injury, poly（lactic-co-glycolic acid） conduits exhibited good intensity, elasticity and plasticity, indicating that poly（lactic-co-glycolic acid） conduits are suitable for sciatic nerve injury repair.

Treatment with acetyl-L-carnitine exerts a neuroprotective effect in the sciatic nerve following loose ligation:a functional and microanatomical study

Peripheral neuropathies are chronic painful syndromes characterized by allodynia,hyperalgesia and altered nerve functionality.Nerve tissue degeneration represents the microanatomical correlate of peripheral neuropathies.Aimed to improve the therapeutic possibilities,this study investigated the hypersensitivity and the neuromorphological alterations related to the loose ligation of the sciatic nerve in rats.Effects elicited by treatment with acetyl-L-carnitine（ALCAR） in comparison to gabapentin were assessed.Axonal injury,reduction of myelin deposition and accumulation of inflammatory cells were detected in damaged nerve.A decrease of phosphorylated 200-k Da neurofilament（NFP） immunoreactivity and a redistribution in small clusters of myelin basic like-protein（MBP） were observed in ipsilateral nerves.Treatment with ALCAR（100 mg/kg intraperitoneally-i.p.） and gabapentin（70 mg/kg i.p.） administered bis in die for 14 days induced a significant pain relieving effect.ALCAR,but not gabapentin,significantly countered neuromorphological changes and increased axonal NFP immunoreactivity.These findings indicate that both ALCAR and gabapentin significantly decreased the hypersensitivity related to neuropathic lesions.The observation of the positive ALCAR effect on axonal and myelin sheath alterations in damaged nerve supports its use as neurorestorative agent against neuropathies through mechanism（s） consistent to those focused in this study.

Complement components of nerve regeneration conditioned fluid influence the microenvironment of nerve regeneration

Nerve regeneration conditioned fluid is secreted by nerve stumps inside a nerve regeneration chamber.A better understanding of the proteinogram of nerve regeneration conditioned fluid can provide evidence for studying the role of the microenvironment in peripheral nerve regeneration.In this study,we used cylindrical silicone tubes as the nerve regeneration chamber model for the repair of injured rat sciatic nerve.Isobaric tags for relative and absolute quantitation proteomics technology and western blot analysis confirmed that there were more than 10 complement components（complement factor I,C1q-A,C1q-B,C2,C3,C4,C5,C7,C8β and complement factor D） in the nerve regeneration conditioned fluid and each varied at different time points.These findings suggest that all these complement components have a functional role in nerve regeneration.

Aminooxyacetic acid improves learning and memory in a rat model of chronic alcoholism

Chronic alcoholism seriously damages the central nervous system and leads to impaired learning and memory.Cell damage in chronic alcoholism is strongly associated with elevated levels of hydrogen sulfide（H2S） and calcium ion overload.Aminooxyacetic acid is a cystathionine-β-synthase activity inhibitor that can reduce H2S formation in the brain.This study sought to observe the effect of aminooxyacetic acid on learning and memory in a chronic alcoholism rat model.Rats were randomly divided into three groups.Rats in the control group were given pure water for 28 days.Rats in the model group were given 6% alcohol for 28 days to establish an alcoholism rat model.Rats in the aminooxyacetic acid remedy group were also given 6% alcohol for 28 days and were also intraperitoneally injected daily with aminooxyacetic acid（5 mg/kg） from day 15 to day 28.Learning and memory was tested using the Morris water maze test.The ultrastructure of mitochondria in the hippocampus was observed by electron microscopy.H2S levels in the hippocampus were measured indirectly by spectrophotometry,and ATPase activity was measured using a commercial kit.The expression of myelin basic protein was determined by immunohistochemistry and western blotting.Compared with the control group,latency and swimming distance were prolonged in the navigation test on days 2,3,and 4 in the model group.In the spatial probe test on day 5,the number of platform crosses was reduced in the model group.Cristae cracks,swelling or deformation of mitochondria appeared in the hippocampus,the hippocampal H2S level was increased,the mitochondrial ATPase activity was decreased,and the expression of myelin basic protein in the hippocampus was down-regulated in the model group compared with the control group.All the above indexes were ameliorated in the aminooxyacetic acid remedy group compared with the model group.These findings indicate that aminooxyacetic acid can improve learning and memory in a chronic alcoholism rat model,which may be associated with reduction of hippocampal H2S level and mitochondrial ATPase activity,and up-regulation of myelin basic protein levels in the hippocampus.

Pulsed radiofrequency alleviated neuropathic pain by down-regulating the expression of substance P in chronic constriction injury rat model

Background:Pulsed radiofrequency(PRF),as a non-invasive treatment of neuropathic pain(NP),has been widely administered clinically.Previous studies have shown that PRF has the potential to improve hyperalgesia in animal models of NP.However,there have been few reports to clarify whether the mechanism of PRF treatment of NP involves intervention in the expression of substance P(SP).Therefore,this study administered PRF treatment to chronic constriction injury(CCI)model rats and observed the sciatic nerve mechanical pain threshold and SP expression in the spinal cord to explore the mechanism of PRF treatment.Methods:A total of 96 Sprague-Dawley rats were randomly divided into the sham-surgery-sham-treatment group(S-S group),the sham-surgery-PRF group(S-P group),the CCI-sham-treatment group(C-S group),and the CCI-PRF group(C-P group).The C-S group and the C-P group underwent sciatic nerve CCI,while the other groups received a sham operation.At 14 days after the operation,the C-P group and the S-P group were treated with PRF for 300 s.We recorded the hindpaw withdrawal threshold(HWT)and the thermal withdrawal latency(TWL)of rats in the various groups at baseline,before treatment(0 days),and at 1,7,14,and 28 days after treatment.L4 to L6 spinal cord tissues were taken before treatment(0 days)and 1,7,14,and 28 days after treatment.The transcription and translation of SP were measured by quantitative polymerase chain reaction and Western blotting,respectively.Results:The HWT and the TWL in the C-P group 28 days after PRF treatment were significantly higher than those in the C-S group(95%confidence interval[CI]:5.84–19.50,P<0.01;95%CI:2.58–8.69,P=0.01).The expression of SP in the C-P group 28 days after PRF treatment was significantly lower than that in the C-S group(95%CI:1.17–2.48,P<0.01).Conclusions:PRF may alleviate CCI-induced NP by down-regulating the expression of SP in the spinal cord of CCI model rats.

Estrogen affects neuropathic pain through upregulating N-methyl-D-aspartate acid receptor 1 expression in the dorsal root ganglion of rats

Estrogen affects the generation and transmission of neuropathic pain,but the specific regulatory mechanism is still unclear.Activation of the N-methyl-D-aspartate acid receptor 1（NMDAR1） plays an important role in the production and maintenance of hyperalgesia and allodynia.The present study was conducted to determine whether a relationship exists between estrogen and NMDAR1 in peripheral nerve pain.A chronic sciatic nerve constriction injury model of chronic neuropathic pain was established in rats.These rats were then subcutaneously injected with 17β-estradiol,the NMDAR1 antagonist D（-）-2-amino-5-phosphonopentanoic acid（AP-5）,or both once daily for 15 days.Compared with injured drug na？ve rats,rats with chronic sciatic nerve injury that were administered estradiol showed a lower paw withdrawal mechanical threshold and a shorter paw withdrawal thermal latency,indicating increased sensitivity to mechanical and thermal pain.Estrogen administration was also associated with increased expression of NMDAR1 immunoreactivity（as assessed by immunohistochemistry） and protein（as determined by western blot assay） in spinal dorsal root ganglia.This 17β-estradiol-induced increase in NMDAR1 expression was blocked by co-administration with AP-5,whereas AP-5 alone did not affect NMDAR1 expression.These results suggest that 17β-estradiol administration significantly reduced mechanical and thermal pain thresholds in rats with chronic constriction of the sciatic nerve,and that the mechanism for this increased sensitivity may be related to the upregulation of NMDAR1 expression in dorsal root ganglia.

Radial shock wave therapy in the treatment of chronic constriction injury model in rats： a preliminary study

Background Pain physicians pay close attention to neuropathic pain （NP）,since there is currently no ideal treatment.Radial shock wave therapy （RSWT） is a noninvasive treatment to chronic pain of soft tissue disorders.So far,there is no information on the use of RSWT for the treatment of NP.Therefore we observe the effects of RSWT on a NP model induced by chronic constriction injury （CCI） in rats.Methods Four different energy densities （1.0,1.5,2.0 and 2.5 bar） RSWT administered as a single session or repeated sessions in rats with NP induced by CCI of the sciatic nerve.The analgesic effect was assessed by measuring mechanical withdrawal threshold （MWT） and thermal withdrawal latency （TWL）.The safety was assessed through calculating sciatic functional index （SFI）.Results MWT and TWL increased after a single session of RSWT from day 1 to day 5 but retumed to baseline levels by day 10.Following repeated sessions of RSWT,both the MWT and TWL were significantly higher than NP group （P ＜ 0.01）for at least 4 weeks.In addition,no significant changes of SFI were observed in any groups after repeated sessions of RSWT and no increased pain or other side effects in any animals.Conclusions A single session of RSWT is rapidly effective in the treatment of CCI,but the efficacy maintained in a short period.However,repeated sessions of RSWT have prolonged efficacy.

Molecular mechanisms underlying the effects of acupuncture on neuropathic pain

Acupuncture has been used to treat neuropathic pain for a long time, but its mechanisms of action remain unknown. In this study, we observed the effects of electroacupuncture and manual acu- puncture on neuropathic pain and on ephrin-B/EphB signaling in rats models of chronic constriction injury-induced neuropathic pain. The results showed that manual acupuncture and electroacu- puncture significantly reduced mechanical hypersensitivity following chronic constriction injury, es- pecially electroacupuncture treatment. Real-time PCR results revealed that ephrin-B1/B3 and EphB1/B2 mRNA expression levels were significantly increased in the spinal dorsal horns of chronic constriction injury rats. Electroacupuncture and manual acupuncture suppressed the high expres- sion of ephrin-B1 mRNA, and elevated EphB3/B4 mRNA expression. Electroacupuncture signifi- cantly enhanced the mRNA expression of ephrin-B3 and EphB3/B6 in the dorsal horns of neuro- pathic pain rats. Western blot results revealed that electroacupuncture in particular, and manual acupuncture, significantly up-regulated ephrin-B3 protein levels in rat spinal dorsal horns. The re- sults of this study suggest that acupuncture could activate ephrin-B/EphB signaling in neuropathic pain rats and improve neurological function.

Effect of Pulsed Radiofrequency on Rat Sciatic Nerve Chronic Constriction Injury： A Preliminary Study

Background：Pulsed radiofrequency （PRF） application to the dorsal root ganglia can reduce neuropathic pain （NP） in animal models,but the effect of PRF on damaged peripheral nerves has not been examined.We investigated the effect of PRF to the rat sciatic nerve （SN） on pain-related behavior and SN ultrastructure following chronic constriction injury （CCI）.Methods：The analgesic effect was measured by hindpaw mechanical withdrawal threshold （MWT） and thermal withdrawal latency （TWL）.Twenty rats with NP induced by ligating the common SN were then randomly divided into a PRF treatment group and a sham group.The contralateral SN served as a control.The MWT and TWL were determined again 2,4,6,8,10,12,and 14 days after the PRF or sham treatment.On day 14,ipsilateral and contralateral common SNs were excised and examined by electron microscopy.Results：Ipsilateral MWT was significantly reduced and TWL significantly shorter compared to the contralateral side 14 days after CCI （both P =0.000）.In the PRF group,MWT was significantly higher and TWL significantly longer 14 days after the PRF treatment compared to before PRF treatment （both P =0.000）,while no such difference was observed in the sham group （P ＞ 0.05）.Electron microscopy revealed extensive demyelination and collagen fiber formation in the ipsilateral SN of sham-treated rats but sparse demyelination and some nerve fiber regrowth in the PRF treatment group.Conclusions：Hyperalgesia is relieved,and ultrastructural damage ameliorated after direct PRF treatment to the SN in the CCI rat model of NP.

Recent evidence for activity-dependent initiation of sympathetic sprouting and neuropathic pain

Traumatic injury or inflammatory irritation of the peripheral nervous system often leads to persistent pathophysiological pain states.It has been well-documented that,after peripheral nerve injury or inflammation,functional and anatomical alterations sweep over the entire peripheral nervous system including the peripheral nerve endings,the injured or inflamed afferent fibers,the dorsal root ganglion(DRG),and the central afferent terminals in the spinal cord.Among all the changes,ectopic discharge or spontaneous activity of primary sensory neurons is of great clinical interest,as such discharges doubtless contribute to the develop-ment of pathological pain states such as neuropathic pain.Two key sources of abnormal spontaneous activity have been identified following peripheral nerve injury:the injured afferent fibers(neuroma) leading to the DRG,and the DRG somata.The purpose of this review is to provide a global account of the abnormal spontaneous activity in various animal models of pain.Particular attention is focused on the consequence of peripheral nerve injury and localized inflammation.Further,mechanisms involved in the generation of spontaneous activity are also reviewed;evidence of spontaneous activity in contributing to abnormal sympathetic sprouting in the axotomized DRG and to the initiation of neuropathic pain based on new findings from our research group are discussed.An improved understanding of the causes of spontaneous activity and the origins of neuropathic pain should facilitate the development of novel strategies for effective treatment of pathological pain.

Repair and regeneration of peripheral nerve injuries that ablate branch points

The peripheral nervous system has an extensive branching organization, and peripheral nerve injuries that ablate branch points present a complex challenge for clinical repair. Ablations of linear segments of the PNS have been extensively studied and routinely treated with autografts, acellular nerve allografts, conduits, wraps, and nerve transfers. In contrast, segmental-loss peripheral nerve injuries, in which one or more branch points are ablated so that there are three or more nerve endings, present additional complications that have not been rigorously studied or documented. This review discusses:(1) the branched anatomy of the peripheral nervous system,(2) case reports describing how peripheral nerve injuries with branched ablations have been surgically managed,(3) factors known to influence regeneration through branched nerve structures,(4) techniques and models of branched peripheral nerve injuries in animal models, and(5) conclusions regarding outcome measures and studies needed to improve understanding of regeneration through ablated branched structures of the peripheral nervous system.