Functional nerve recovery after bridging a 15 mm gap in rat sciatic nerve with a biodegradable nerve guide.

Recovery of nerve function was evaluated after bridging a 15 mm sciatic nerve gap in 51 rats with a biodegradable poly(DL-lactide-epsilon-caprolactone) nerve guide. Recovery of function was investigated by analysing the footprints, by analysing video recordings of gait, by electrically eliciting the withdrawal reflex, by nerve conduction velocity and by electromyography (EMG). Sensory nerve function recovered as measured by electrostimulation. Motor nerve function partly recovered but electromyograms remained abnormal throughout the study. We conclude that functional reinnervation by regenerating axons occurs after bridging a 15 mm nerve gap with a biodegradable poly(DL-lactide-epsilon-caprolactone) nerve guide, but the walking patterns remain abnormal. Video analysis is a useful tool to record and analyse the walking patterns of rats. Further studies are necessary to investigate the possibility of obtaining selective reinnervation of specific muscles.     

优化法去细胞大鼠神经同种异体移植修复坐骨神经缺损

[目的]以优化法去细胞大鼠神经移植,修复同种异体坐骨神经缺损,观察术后动物的免疫排斥、早期功能恢复及神经再生情况。[方法]以优化去细胞方法处理新鲜取材的成年SD大鼠坐骨神经,移植修复同种异体1.0cm坐骨神经缺损,以自体神经和新鲜异体神经移植为对照,术后1个月行坐骨神经功能指数评价、神经电生理和组织学检查,观察动物在功能恢复、免疫排斥及神经再生方面的情况。[结果]自体神经和去细胞异体神经移植组动物的坐骨神经功能指数无显著差异(P>0.05),大体观察均可见神经连续性良好。电生理检测表明2组动物移植神经均已恢复电传导能力,在传导速度(CV)上无显著差异(P>0.05),但均未达到正常神经水平(P<0.05)。组织学观察则显示2组再生神经纤维均已长入移植段远端。S-100免疫组化显示两者在雪旺氏细胞数、形态和排列等方面无明显差异。2组在CD8 T细胞和巨噬细胞免疫组化染色阳性面积百分比上差异无统计学意义(P>0.05)。计算移植神经中段轴突密度后表明两者无显著差异(P>0.05),但都比正常神经小(P<0.05),比新鲜异体神经移植组大(P<0.05)。[结论]优化去细胞神经移植组与自体神经移植组在免疫排斥、功能恢复及神经再生方面无显著差异。优化法去细胞神经在移植修复同种异体神经缺损时,可以达到免疫耐受,其早期功能恢复和神经再生情况良好,在修复周围神经缺损时可以作为自体神经移植的一种替代疗法。     

无细胞的异体神经修复鼠坐骨神经缺损

目的 通过化学萃取同种异体神经,去除髓鞘和雪旺细胞,形成无细胞基膜管后桥接鼠坐骨神经缺损,研究神经再生效果。方法 正常鼠坐骨神经用非变性生物剂处理后得到无细胞的基膜管,桥接鼠坐骨神经20mm缺损。实验分3组:无细胞基膜管移植组(A组),自体神经移植组(B组)和异体神经移植组(C组)。术后进行肌电图、光镜、电镜及图象分析仪检查。结果 A组再生神经有大量轴突通过移植体,术后2个月电生理检测再生神经的潜伏期及波幅低于B组(P<0.05),术后3个月2组差异无显著意义。髓鞘厚度在术后3个月时亦低于B组,差异有显著意义(P<0.05)。轴突直径及数目两组无差异。C组因无神经再生,结果无法测量。结论 这种无细胞基膜管移植体能支持轴突的生长和雪旺细胞的迁移,是一种良好的神经移植替代材料。     

Functional assessment of sciatic nerve recovery: biodegradable poly (DLLA-epsilon-CL) nerve guide filled with fresh skeletal muscle

The aim of this study was to compare functional peripheral nerve recovery in the rat sciatic nerve model after reconstruction of a 10-mm gap with a biodegradable poly (DLLA-epsilon-CL) nerve guide, as filled with either fresh skeletal muscle or phosphate-buffered saline (PBS). During 24 weeks of recovery, motor and sensory functional evaluation was tested by extensor postural thrust (EPT) and withdrawal reflex latency (WRL), respectively. At the end of the experiment, anesthetized animals were prepared for motor nerve conduction velocity (MNCV) studies, followed by gastrocnemius and soleus muscle weight measurement. Motor functional recovery was greater in the muscle-grafted group, and reached a significant difference from weeks 8-12 (P < 0.05). The results of this investigation suggest that filling a nerve guide with fresh skeletal muscle induces faster maturation of regenerated nerve fibers in comparison with traditional tubular repair.     

犬化学去细胞神经同种异体移植的实验研究

目的以化学去细胞同种坐骨神经移植修复犬坐骨神经的长段缺损，观察其功能恢复及神经再生。方法15犬分成去细胞同种神经组(实验组)6犬、自体神经组(对照组Ⅰ)6犬、新鲜同种神经组(对照组Ⅱ)3犬。右侧坐骨神经造成5．0cm长缺损，以上述三种移植物桥接修复。术后6个月行步态分析、神经电生理及神经再生观察。结果实验组和对照组Ⅰ在运动功能恢复，踝关节运动步态，小腿二头肌运动诱发电位、感觉诱发电位，移植段内新生轴突、血管及雪旺细胞，远端胫神经内有髓神经纤维及靶肌肉运动终板等方面非常相似。对照组Ⅱ神经功能始终无恢复，移植段被吸收。结论化学去细胞同种神经移植物修复犬粗大长段神经缺损时不会被宿主排斥和吸收，其近期功能恢复及神经再生与自体神经移植无明显差别。     

Large-area irradiated low-level laser effect in a biodegradable nerve guide conduit on neural regeneration of peripheral nerve injury in rats

This study used a biodegradable composite containing genipin-cross-linked gelatin annexed with β-tricalcium phosphate ceramic particles (genipin-gelatin-tricalcium phosphate, GGT), developed in a previous study, as a nerve guide conduit. The aim of this study was to analyse the influence of a large-area irradiated aluminium-gallium-indium phosphide (AlGaInP) diode laser (660 nm) on the neural regeneration of the transected sciatic nerve after bridging the GGT nerve guide conduit in rats. The animals were divided into two groups: group 1 comprised sham-irradiated controls and group 2 rats underwent low-level laser (LLL) therapy. A compact multi-cluster laser system with 20 AlGaInP laser diodes (output power, 50 mW) was applied transcutaneously to the injured peripheral nerve immediately after closing the wound, which was repeated daily for 5 min for 21 consecutive days. Eight weeks after implantation, walking track analysis showed a significantly higher sciatic function index (SFI) score (P < 0.05) and better toe spreading development in the laser-treated group than in the sham-irradiated control group. For electrophysiological measurement, both the mean peak amplitude and nerve conduction velocity of compound muscle action potentials (CMAPs) were higher in the laser-treated group than in the sham-irradiated group. The two groups were found to be significantly different during the experimental period (P < 0.005). Histomorphometric assessments revealed that the qualitative observation and quantitative analysis of the regenerated nerve tissue in the laser-treated group were superior to those of the sham-irradiated group. Thus, the motor functional, electrophysiologic and histomorphometric assessments demonstrate that LLL therapy can accelerate neural repair of the corresponding transected peripheral nerve after bridging the GGT nerve guide conduit in rats.     

Functional recovery after facial and sciatic nerve crush injury in the rat

OBJECTIVES: To systematically record rat facial nerve recovery following crush injury to the main trunk with respect to ocular and vibrissial function and to compare the rates of facial and sciatic nerve recovery from crush injury in the same animals. This serves as a means of validating the functional parameters of facial nerve recovery against the well-known measure of hind limb function, the Sciatic Function Index. METHODS: The main trunk of the facial nerve and the proximal segment of the sciatic nerve were exposed in all animals. Both nerves were subjected to standardized crush injury and subsequent daily functional testing. After a plateau of functional recovery was achieved, the animals were killed, and the distances between the sites of injury and the end musculature were measured, which allowed determination and comparison of recovery rates in both systems. RESULTS: All crush injuries resulted in loss of electrical conductivity, as proven by intraoperative proximal nerve stimulation. Recovery of ocular and vibrissial motor function occurred starting at postoperative day (POD) 9 and continuing through POD 20. Hind limb function returned later (POD 14-34); however, when corrected for distance, the sciatic recovery rate (2.26 mm/d) appeared to match that of the facial nerve (1.5-2.4 mm/d). CONCLUSIONS: Recovery after facial nerve crush injury follows a predictable time course, and the rate of recovery is consistent with that of sciatic nerve injury. Return of the blink reflex, loss of vibrissial fibrillations, and return of vibrissial sweeping function appear to be internally consistent functional measures of facial recovery. These quantitative measures will be useful for future facial nerve manipulation studies.     

Comparison of assessment tools to score recovery of function after repair of traumatic lesions of the median nerve

In this paper the recovery after repair of the median nerve has been used to compare different assessment tools for evaluation of peripheral nerve function: touch (moving 2-point discrimination (2PD); Semmes-Weinstein (SW) monofilament, motor (Medical Research Council (MRC) scale), combined motor and sensory (Dellon modification of the Moberg pick up test; Moberg Recognition test), and pain (visual analogue scale; pinprick-test). The mean (SD) age of our 28 patients was 28 (12) years. The mean (SD) follow-up period was 5 years, 2 months (2 years, 8 months). On the operated side three patients (11%) had a moving 2PD of less than 4 mm. The results of the moving 2PD were compared with those of the SW monofilaments, but with a poor correlation. The MRC score correlated well with opposition movement of the thumb and muscle wasting (p<0.01). We recommend a number of tests to evaluate (the chronological return of) peripheral nerve function.     

Functional recovery of sciatic nerve through inside-out vein graft in rats

Objective: Present study aimed at further comprehensive functional, histomorphometrical and immunohistochemical assessment of peripheral nerve regeneration using rat sciatic nerve transection model.Methods: The 10-mm rat sciatic nerve gap was created in rats. In control group nerve stumps were sutured to adjacent muscle and in treatment group the gap was bridged using an inside-out vein graft. In sham-operated group the nerve was manipulated and left intact. All animals underwent walking track analysis test 4, 8, and 12 weeks after surgery.Subsequently, muscle mass measurement was performed to assess reenervation, histological examination to observe the sciatic nerve regeneration morphologically and immunohistochemistry to detect Schwann cells using anti S-100. Results were analyzed using a factorial ANOVA with two between-subjects factors. Bonferroni test for pairwise comparisons was used to examine the effect of treatments.Results: Functional analysis ofmyelinated nerve fibers showed that nerve function improved significantly in the time course in treatment group. However, quantitative morphometrical analysis of myelinated nerve fibers showed that there was no significant difference between 8 and 12 weeks in treatment group. Muscle weight ratio was bigger and weight loss of the gastrocnemius muscle was ameliorated by inside-out vein grafting. The position of positive immunohistochemical reactions further implied that regenerated axons and Schwann cell-like cells existed after vein grafting was performed, and was accompanied by the process of myelination and structural recovery of regenerated nerves.Conclusion: Functional analysis of peripheral nerve repair is far more reliable than quantitative morphometrical analysis     

Functional assessment after sciatic nerve injury in a rat model

The aim of this study is to evaluate the effectiveness of Sciatic Function Index (SFI) and Basso, Beattie, and Bresnahan (BBB) Locomotor Rating in assessing peripheral nerve injuries. SFI is a standard method for evaluating crush and transected peripheral nerve injuries, likewise BBB for spinal cord injury. Models of chronic nerve compression (CNC), crush, and transection injury were created on Sprague‐Dawley rats and functional outcomes were measured using BBB and SFI at 1‐week interval for 6 weeks. All injury models showed high correlation between SFI and BBB scores. With crush injury, the SFI showed near complete recovery while BBB showed residual deficits 6 weeks after injury. Both the BBB and SFI were unable to detect motor deficits in 6‐week CNC animals. The BBB score should be considered as an adjunct in evaluating peripheral nerve recovery and may be more sensitive in detecting residual deficits than SFI after crush‐type injuries. © 2009 Wiley‐Liss, Inc. Microsurgery, 2009.     

Functional recovery after discharge in enhanced recovery video-assisted thoracoscopic lobectomy: a pilot prospective cohort study

Little is known about functional recovery following patient discharge in an established enhanced recovery programme after video-assisted thoracoscopic lobectomy. We conducted a single-centre pilot prospective observational cohort study. We hypothesised that patients achieved early functional recovery after discharge. A total of 32 patients aged ≥ 18 years were enrolled. A digital device was used for objective activity measurements, and patient-reported outcomes were collected as subjective measurements. Primary outcomes were the difference in physical activity; sleep duration; pain; fatigue; and average quality of life scores between pre-operative baseline and 7 days following discharge. The secondary outcome was the reason for reduced daily activity during the first 7 days after discharge. Median (IQR [range]) length of stay was 3 (2–5 [1–13]) days. Up to post-discharge day 7, total, lower intensity and moderate-to-vigorous activities were lower than pre-operative activity (p < 0.001; p = 0.005 and p = 0.027, respectively). Numerical rating scale (0–10) pain scores increased postoperatively at rest (mean difference 1.2, p < 0.001) and during walking (mean difference 1.4, p < 0.001). Fatigue assessed by the Christensen Fatigue Scale (1–10) was also increased postoperatively (mean difference 1.7, p = 0.001). There was a reduction in quality of life scores, while sedentary activity and sleep duration were unchanged postoperatively. Dominant reasons for not recovering daily activity included fatigue in 43% and pain in 33% of patients. Despite compliance with an enhanced recovery programme with a median length of hospital stay of 3 days after video-assisted thoracoscopic lobectomy, functional recovery was not achieved within 7 days after hospital discharge. Reduction in postoperative pain and fatigue are important factors to enhance functional recovery.     

Effects of collagen nerve guide on neuroma formation and neuropathic pain in a rat model

BACKGROUND: Posttraumatic neuroma formation is a major cause of neuropathic pain that can occur after elective surgery, amputation, or trauma. This study examined the use of biosynthetic collagen nerve guides to prevent the development of posttraumatic neuromas. METHODS: Collagen nerve guides were applied after neurectomy in a rat sciatic nerve model in an effort to stimulate linear neuronal outgrowth and reduce random axon sprouting. Animals were monitored for evidence of neuropathic pain--autotomy scores were recorded for 8 weeks posttransection--after which proximal stumps were excised and processed for histologic analyses. RESULTS: Moderate to severe autotomy was observed in 88% (7 of 8) of the control (neurectomy) animals. In contrast, 13% (1 of 8) of animals receiving collagen nerve guides developed autotomy, which was significantly less than controls (P < .01). Qualitative analyses of neurofilament and Schwann cell-labeled nerve sections showed a significant enhancement in Schwann cell migration away from the proximal stump and advanced linear axonal regrowth in the collagen nerve guide-treated animals. CONCLUSIONS: Collagen nerve guides alter the regrowth of transected nerves and reduce the severity of symptoms associated with neuropathic pain.     

嗅鞘细胞促进大鼠坐骨神经损伤后神经功能恢复

目的研究嗅鞘细胞（Olfactory ensheathing cells,OECs）移植对大鼠坐骨神经损伤后神经再生恢复的作用。方法取SD大鼠40只随机分成对照生理盐水（SAL）组和实验（OECs）组,予离断坐骨神经后直接予神经外膜缝合修复,在神经缝合处周围充填可吸收的明胶海绵,SAL组和OECs组明胶海绵内分别给予SAL和体外培养纯化的OECs;术后4、12周分别行大体观察,电生理检查和组织学检查。结果术后4、12周,SAL组和OECs组修复神经均有不同程度恢复,OECs组在运动神经传导速度、神经肌肉动作电位幅度和直径数据上有优于SAL组,但统计学上未见明显差异,而在有髓神经纤维数目方面明显优于SAL组（P〈0.05）。结论在本实验条件下,嗅鞘细胞（OECs）对大鼠坐骨神经损伤后的神经功能恢复有部分的促进作用。     

Claudin 14/15 play important roles in early wallerian degeneration after rat sciatic nerve injury

PurposeWallerian degeneration (WD) is an antegrade degenerative process distal to peripheral nerve injury. Numerous genes are differentially regulated in response to the process. However, the underlying mechanism is unclear, especially the early response. We aimed at investigating the effects of sciatic nerve injury on WD via CLDN 14/15 interactions in vivo and in vitro.MethodsUsing the methods of molecular biology and bioinformatics analysis, we investigated the molecular mechanism by which claudin 14/15 participate in WD. Our previous study showed that claudins 14 and 15 trigger the early signal flow and pathway in damaged sciatic nerves. Here, we report the effects of the interaction between claudin 14 and claudin 15 on nerve degeneration and regeneration during early WD.ResultsIt was found that claudin 14/15 were upregulated in the sciatic nerve in WD. Claudin 14/15 promoted Schwann cell proliferation, migration and anti-apoptosis in vitro. PKCα, NT3, NF2, and bFGF were significantly upregulated in transfected Schwann cells. Moreover, the expression levels of the β-catenin, p-AKT/AKT, p-c-jun/c-jun, and p-ERK/ERK signaling pathways were also significantly altered.ConclusionClaudin 14/15 affect Schwann cell proliferation, migration, and anti-apoptosis via the β-catenin, p-AKT/AKT, p-c-jun/c-jun, and p-ERK/ERK pathways in vitro and in vivo. The results of this study may help elucidate the molecular mechanisms of the tight junction signaling pathway underlying peripheral nerve degeneration.     

Vascular endothelial growth factor promotes peripheral nerve regeneration after sciatic nerve transection in rat

Objective：To evaluate the local effect of vascular endothelial growth factor （VEGF） on transected sciatic nerve regeneration.Methods：Sixty male white Wistar rats were divided into four experimental groups randomly （n=15）.In transected group the left sciatic nerve was transected and the stump was fixed to adjacent muscle.In treatment group the defect was bridged using a silicone graft filled with 10μL VEGF.In silicone group the graft was filled with phosphate-buffered saline.In sham-operated group the sciatic nerve was exposed and manipulated.Each group was subdivided into three subgroups with five animals in each and nerve fibers were studied 4,8 and 12 weeks after operation.Results：Behavioral test,functional study of sciatic nerve,gastrocnemius muscle mass and morphometric indices confirmed a faster recovery of regenerated axons in VEGF group than in silicone group （P〈0.05）.In immunohistochemical assessment,reactions to S-100 in VEGF group were more positive than that in silicone group.Conclusion：Local administration of VEGF will improve functional recovery and morphometric indices of sciatic nerve.     

Implantation of Schwann cells in rat tendon autografts as a model for peripheral nerve repair: Long term effects on functional recovery

Cultured Schwann cells in tendon autografts for nerve repair improve the early phase of nerve regeneration in rat sciatic nerves as judged by the rate of axonal outgrowth. We tested the long-term effects on functional recovery using measurements of muscle force, the number of axons and myelination, using morphometry. In addition, we recorded wet weight of the gastrocnemius muscle. Schwann cell cultures were prepared from predegenerated nerves. Ten and 15mm defects in rat sciatic nerves were bridged using bilateral tendon autografts with Schwann cell-seeded tendon autografts on one side, and untreated tendon autografts on the other. Animals were evaluated at six and 12 weeks, respectively. At six weeks, myelination, as judged by G-ratio (ratio of axonal diameter to diameter of nerve fibres), was significantly increased in tendon autografts pretreated with Schwann cells in 10mm defects. No such difference was seen in the 15 mmdefects. We found no difference in functional recovery, other morphometric variables, or muscle weight between the two grafts. We conclude that early effects on nerve regeneration using transplantation of cultured Schwann cells in rat sciatic nerves are temporary. Other strategies are necessary to obtain lasting effects on functional recovery.