欢迎访问《解剖学报》官方网站！今天是
English
小鼠神经缺损管式修复后长时程功能恢复与再生评价
Long-term evaluation of functional recovery and nerve regeneration following tubulation repair of nerve defects in mice
目的 了解壳聚糖/聚乳酸-乙醇酸共聚体（PLGA）人工神经移植物修复小鼠神经缺损后神经功能长时程恢复水平与再生神经成熟度。方法 采用人工神经移植物桥接修复小鼠坐骨神经缺损(n=6)，以自体神经修复(n=6)和缺损组(n=6)为对照，术后1年采用热痛阈测定、电生理学、激光多普勒血流检测评定神经功能，采用靶肌湿重比、组织学和电子显微镜等技术综合评定神经重支配和再生神经成熟度。 结果 人工神经移植物组足底痛觉反应潜伏期、神经源性血管扩张程度、腓肠肌复合肌动作电位(CMAPs)波幅和潜伏期、靶肌湿重比、再生轴突数量等指标与自体神经修复组相近，但与健侧相比CMAPs潜伏期较长，髓鞘较薄，轴突直径分布滞后。
结论 人工神经移植物修复小鼠神经缺损术后1年感觉及自主神经功能、再生神经数量和靶肌重支配水平与自体神经修复相当，但再生神经纤维成熟度未达正常。
Objective This study is to identify long-term functional recovery and maturity of regenerated nerve fibers after repairing mouse nerve defects with chitosan/polylactide-co-polyglycolide artificial nerve grafts (CPANGs). Methods Mouse sciatic nerve defects, 2mm in length, were bridged by CPANGs (n=6), with nerve autograft (n=6) and nerve defect (n=6) as controls. Plantar test, electrophysiological examination and laser Doppler perfusion imaging following nerve crush were carried out at 1 year after repair to assess nerve function recovery, while muscle wet weight ratio, histological assessment and transmission electron microscopy were performed to evaluate nerve re-innervation and maturity of regenerated nerve fibers. Results When compared to the autograft group, the CPANG group did not show statistically significant difference in functional recovery in terms of paw withdrawal latency, neurogenic vasodilatation, amplitude and latency of compound muscle action potentials (CMAPs), wet weight ratio of gastrocnemius and tibialis cranialis muscles, number of myelinated nerve fibers and density of unmyelinated axons. However, both these two repair groups exhibited significantly longer CMAPs latency, thinner myelin sheath and a lagbehind shift of diameter distribution of myelinated axons as compared to the normal control. Conclusion At 1 year after the mouse sciatic nerve defect was repaired by CPANGs, sensory and autonomic nerve function, number of regenerated axons and muscle re-nnervation degree were recovered to the same extent as nerve autografting, but the regenerated nerve fibers were in a state of immaturity.
神经损伤 / 神经修复 / 壳聚糖 / 聚乳酸-乙醇酸共聚体 / 激光多普勒血流检测 / 电生理学 / 小鼠
Nerve injury / Nerve repair / Chitosan / Polylactide-co-polyglycolide / Lacer doppler perfusion imaging / Electrophysiological examinations / Mouse
［1］Gu X, Ding F, Yang Y, et al. Construction of tissue engineered nerve grafts and their application in peripheral nerve regeneration ［J］. Prog Neurobiol, 2011, 93(2): 204-230.
［2］Gu J, Hu W, Deng A, et al. Surgical repair of a 30mm long human median nerve defect in the distal forearm by implantation of a chitosan-PGA nerve guidance conduit ［J］. J Tissue Eng Regen Med, 2012, 6(2): 163-168.
［3］Yao J, Shi W, Yuan Y, et al. Effect of the artificial nerve graft on nerve regeneration in rat sciatic nerve of two months defect model ［J］. Acta Anantomica Sinica, 2007, 38(5): 505-510. (in Chinese)
姚健, 施伟, 袁颖, 等. 人工组织神经移植物对大鼠陈旧性坐骨神经缺损(60天)的修复作用 ［J］. 解剖学报, 2007, 38(5): 500-510.
［4］Wang X, Hu W, Cao Y, et al. Dog sciatic nerve regeneration across a 30-mm defect bridged by a chitosan/PGA artificial nerve graft ［J］. Brain, 2005, 128(Pt 8): 1897-1910.
［5］Hu N, Wu H, Xue C, et al. Long-term outcome of the repair of 50mm long median nerve defects in rhesus monkeys with marrow mesenchymal stem cells-containing, chitosan-based tissue engineered nerve grafts ［J］. Biomaterials, 2013, 34(1): 100-111.
［6］Yang Y, Ding F, Wu J, et al. Development and evaluation of silk fibroin-based nerve grafts used for peripheral nerve regeneration ［J］. Biomaterials, 2007, 28(36): 5526-5535.
［7］Yang Y, Yuan X, Ding F, et al. Repair of rat sciatic nerve gap by a silk fibroin-based scaffold added with bone marrow mesenchymal stem cells ［J］. Tissue Eng Part A, 2011, 17(1718): 2231-2244.
［8］Hu W, Gu XS. Evaluation of peripheral nerve regeneration outcome in experimental settings ［J］. Chinese Journal of Microsurgery, 2012, 10(35): 435-439. (in Chinese)
胡文, 顾晓松. 周围神经损伤动物模型神经再生效果的评价 ［J］. 中华显微外科杂志, 2012, 355(5): 435-439.
［9］Shen ZhY, Gu TW, Hu W. Time-lapse LDPI analysis of skin territory following sciatic nerve transection in rats ［J］. Acta Universitatis Medicinalis Nanjing (Natural Science), 2013, 33(6): 779-783. (in Chinese)
沈钟一, 顾天文, 胡文. 大鼠坐骨神经切断后足底皮肤血流灌注变化研究 ［J］. 南京医科大学学报(自然科学版), 2013, 33(6): 779-783.
［10］Hu W, Liu D, Zhang Y, et al. Neurological function following intra-neural injection of fluorescent neuronal tracers in rats ［J］. Neural Regen Res, 2013, 8(14): 1253-1261.
［11］Korte N, Schenk HC, Grothe C, et al. Evaluation of periodic electrodiagnostic measurements to monitor motor recovery after different peripheral nerve lesions in the rat ［J］. Muscle Nerve, 2011, 44(1): 63-73.
［12］Hashimoto T, Suzuki Y, Kitada M, et al. Peripheral nerve regeneration through alginate gel: analysis of early outgrowth and late increase in diameter of regenerating axons ［J］. Exp Brain Res, 2002, 146(3): 356-368.
［13］Fricker FR, Antunes-Martins A, Galino J, et al. Axonal neuregulin 1 is a rate limiting but not essential factor for nerve remyelination ［J］. Brain, 2013, 136(Pt 7): 2279-2297.
［14］Fricker FR, Lago N, Balarajah S, et al. Axonally derived neuregulin-1 is required for remyelination and regeneration after nerve injury in adulthood ［J］. J Neurosci, 2011, 31(9): 3225-3233.
［15］Michailov GV, Sereda MW, Brinkmann BG, et al. Axonal neuregulin-1 regulates myelin sheath thickness ［J］. Science, 2004, 304(5671): 700-703.
［16］Morton PD, Johnstone JT, Ramos AY, et al. Nuclear factor-kappa B activation in Schwann cells regulates regeneration and remyelination ［J］. Glia, 2012, 60(4): 639-650.
［17］Napoli I, Noon LA, Ribeiro S, et al. A central role for the ERK-signaling pathway in controlling Schwann cell plasticity and peripheral nerve regeneration in vivo ［J］. Neuron, 2012, 73(4): 729-742.
［18］Yang DP, Kim J, Syed N, et al. p38 MAPK activation promotes denervated Schwann cell phenotype and functions as a negative regulator of Schwann cell differentiation and myelination ［J］. J Neurosci, 2012, 32(21): 7158-7168.
［19］Stassart RM, Fledrich R, Velanac V, et al. A role for Schwann cell-derived neuregulin-1 in remyelination ［J］. Nat Neurosci, 2013, 16(1): 48-54.
［20］Hu W, Yang M, Chang J, et al. Laser doppler perfusion imaging of skin territory to reflect autonomic functional recovery following sciatic nerve autografting repair in rats ［J］. Microsurgery, 2012, 32(2): 136-143.
［21］McLean J, Batt J, Doering LC, et al. Enhanced rate of nerve regeneration and directional errors after sciatic nerve injury in receptor protein tyrosine phosphatase sigma knock-out mice ［J］. J Neurosci, 2002, 22(13): 5481-5491.
国家自然科学基金资助项目;江苏省高校自然科学基金;江苏省高校“青蓝工程”项目
/
| 〈 |
|
〉 |
