含神经生长因子的壳聚糖导管桥接周围神经缺损的实验研究

目的 应用含有神经生长因子(NGF)的壳聚糖神经导引管作为神经再生室桥接大鼠坐骨神经缺损，观察其对神经再生的作用。方法 选用Wistat大鼠60只，手术造成右后肢坐骨神经长约15mm的缺损，A组以含有NGF的壳聚糖神经导引管桥接神经缺损，B组则单纯采用壳聚糖导管，分别于术后4、12、24周进行大体及显微解剖观察、组织学检查、电镜观察和神经电生理测定。结果 A组在促进神经再生、加快血管化进程、再生神经纤维排列规律化、提高再生神经髓鞘化、加速再生神经功能重建等方面均优于B组。结论 壳聚糖神经导引管可以为大鼠坐骨神经再生提供一个良好的再生微环境，NGF对神经再生有显促进作用。     

1102/异种神经脱细胞移植物桥接大鼠坐骨神经缺损

目的　探讨异种神经脱细胞移植物桥接大鼠坐骨神经缺损后的神经再生及其再生过程中免疫排斥反应。 方法　用脱细胞兔周围神经作为移植物桥接大鼠坐骨神经1cm缺损;术后3、5、8、11、15天检测血液中淋巴细胞占白细胞百分比;3个月后取移植物及腓肠肌,用甲苯胺蓝、乙酰胆碱酯酶(AchE)、琥珀酸脱氢酶(SDH)组化染色,光、电镜观察神经再生及腓肠肌运动终板的恢复情况。　 结果　术后大鼠血液中淋巴细胞占白细胞的百分比与正常大鼠相比较无显著性差异,3个月后大鼠术侧下肢足趾能分开,行走时后蹬动作有力,针刺足底有逃避反应,桥接物内见有大量再生的坐骨神经纤维,腓肠肌肌纤维上见有呈AchE阳性的运动终板和神经纤维。　 结论　异种神经脱细胞移植物桥接大鼠坐骨神经缺损具有促进其再生的作用。     

联合应用甲壳素导管与IGF-1对大鼠坐骨神经再生的研究

本实验探讨联合应用甲壳素导管与IGF-1对大鼠坐骨神经再生功能的影响,为治疗周围神经损伤提供科学依据。1实验方法健康雄性SD大鼠48只,随机分为3组,每组16只,制备双侧大鼠坐骨神经缺损模型(神经缺损10mm),以可吸收甲壳素神经导管桥接神经两断端形成神经再生室。A组为空白对照组:神经再生室内注入等渗生理盐水20μL;B组为对照组:神经再生室内注入明胶溶液20μL;C组为实验组:神经再生室内注入IGF-1凝胶20μL。术后4周、8周收集标本并观察再生神经的大体形态,检测神经传导速度,并进行特殊染色,S-100免疫组化检测,使用统计软件SPSS 17.0进行统计学分析。     

无细胞神经支架复合骨髓间充质干细胞构建组织工程人工神经修复坐骨神经缺损*☆

背景：作者已经成功制备了无细胞神经移植物,并且复合骨髓间充质干细胞构建组织工程人工神经桥接大鼠坐骨神经缺损。 目的：无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经修复大鼠坐骨神经缺损后运动功能的恢复。 方法：成年雄性SD大鼠构建大鼠坐骨神经15 mm缺损模型,分别应用组织工程人工神经、组织工程神经支架或自行神经桥接坐骨神经缺损。桥接后20周再生神经电生理学测定,手术侧胫骨前肌湿质量、腓肠肌组织学及透视电镜分析。 结果与结论：桥接20周后,组织工程人工神经与自体神经移植组胫骨前肌湿质量比较,差异无显著性意义(P > 0.05),神经干传导速度为(30.56±2.15)m/s。结果提示,无细胞神经移植物复合骨髓间充质干细胞构建的组织工程人工神经桥接大鼠坐骨神经缺损后,可以促进再生神经运动功能的恢复。     

外源性神经生长因子诱导下自体静脉桥接修复神经缺损

背景：采用自体神经游离移植修复神经缺损效果比较理想,但有其弊端。为此寻求一种更佳修复神经缺损的治疗方法。 目的：验证及外源性神经生长因子诱导下自体静脉桥接神经缺损对神经再生的影响。 方法：采用Wistar大鼠建立周围神经缺损模型。随机将大鼠分为3组。实验组采用自体静脉桥接并注入神经生长因子;对照组采用自体静脉桥接并注入生理盐水;标准组采用自体神经桥接。分别于术后1,3个月,对实验动物进行活体观察,电生理检测及组织学检测。 结果与结论：3组实验动物均有神经再生及修复表现,但程度不同。实验组失神经表现恢复的较对照组早,电生理检测运动神经传导速度快,组织学检查再生神经纤维数量及质量明显高于对照组(P < 0.05);与“金标准”的自体神经桥接组比较无显著性意义(P > 0.05)。结果提示采用自体静脉桥接+神经生长因子诱导对周围神经缺损后的再生、修复具有有促进作用,可以使再生神经纤维的数量增加并显著提高再生神经纤维质量。     

无细胞神经移植物复合骨髓间充质干细胞修复大鼠坐骨神经缺损

背景：作者前期试验已成功制备了天然可生物降解的无细胞神经移植物并证明其可促进周围神经再生。 目的：无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经并观察其修复大鼠坐骨神经缺损促进运动功能恢复的效果。 设计、时间及地点：随机对照动物实验，于2008-06/2009-02在辽宁医学院附属第一医院医学组织工程实验室完成。 材料：180~200 g成年健康雄性Wistar大鼠，用于制备无细胞神经移植物，100~120 g成年健康雄性Wistar大鼠，用于制备骨髓间充质干细胞，将骨髓间充质干细胞植入并与无细胞神经支架联合培养构建组织工程人工神经。 方法：180~200 g成年健康雄性SD大鼠60只构建坐骨神经15 mm缺损模型，随机分成3组，每组20只。①实验组采用组织工程人工神经桥接大鼠坐骨神经缺损。②空白对照组采用组织工程神经支架桥接大鼠坐骨神经缺损。③自体神经对照组采用自体神经移植桥接大鼠坐骨神经缺损。 主要观察指标：术后12周通过大体观察、电生理检测、组织学和小腿三头肌湿质量等方法分析评价运动功能恢复情况。 结果：①术后12周，实验组大鼠手术侧足趾可以分开，并且可以支撑着地；实验组大鼠再生神经传导速度与自体神经对照组相比，差异无显著性意义。②术后12周，实验组组织化学染色可见腓肠肌内有呈AchE阳性的运动终板整齐地排列于腓肠肌的中上部形成终板带，经结合镀银染色后可见再生的神经束及发出的分支与运动终板相连。③实验组与自体神经对照组胫骨前肌湿质量比差异无显著性意义。 结论：无细胞神经移植物复合骨髓间充质干细胞桥接大鼠坐骨神经缺损具有促进其运动功能恢复的作用。     

PDLLA／NGF复合导管促进周围神经缺损再生的形态学研究

目的探究聚d,L-乳酸,神经生长因子(PDLLA/NGF)可吸收性复合导管桥接对大鼠坐骨神经缺损再生修复的促进作用,为临床上周围神经损伤再生修复提供实验依据。方法60只SD大鼠随机均分4组:自体神经移植(A组),单纯PDLLA导管(B组),PDLLA加一次性给NGF(C组),复合导管(D组)。制作坐骨神经10 mm缺损,按上述分组桥接,手术后1、2、3月每组各取5只修复处组织。行大体观察、三头肌湿重、光镜观察与图像处理、电镜超微结构观察。结果同时间段组间比较,三头肌恢复率A组高,D组次之,C组较差,B组最差(P<0.05)。组织学观察及图像分析显示,D组再生神经的数、质量都显著优于B、C组(P<0.05),接近A组。电镜观察显示A、D组中段神经纤维密度大、直径大、髓鞘厚,内有发育良好的神经丝和微管。B、C组神经稀、直径小、髓鞘薄、无髓神经相对多。结论PDLLA/NGF复合导管具有良好的组织相容性。用该管桥接修复大鼠受损的坐骨神经,能有效地促进其再生。     

应用肌膜瓣包裹、羊膜管预置聚乳酸-羟基乙酸微丝模拟周围神经再生

背景：周围神经损伤部位的微环境状况是影响神经再生的重要因素之一。周围神经损伤后，良好的神经再生微环境有利于保护受损神经元、促进轴突的有效再生。 目的：应用肌膜瓣包裹、羊膜管预置聚乳酸-羟基乙酸微丝，充填大鼠自体周围神经组织浆模拟周围神经再生微环境，探讨其修复坐骨神经缺损的可行性。 设计、时间及地点：随机对照动物实验，于2006-06/2007-10在广东医学院实验动物中心完成。 材料：清洁级2月龄SD大鼠30只，随机分为实验组、对照组、标准组3组，每组10只，右侧为实验侧，左侧为正常对照侧。取健康、足月、顺产的新鲜胎儿羊膜(产妇知情同意) 制备羊膜基质膜。用医用Vicryl缝线和羊膜基质膜制备聚乳酸-羟基乙酸微丝桥接物。 方法：大鼠切除坐骨神经6.0 mm，自然回缩建立坐骨神经缺损模型。实验组采用带蒂肌膜瓣、人羊膜管预置Vicryl微丝并充填大鼠自体坐骨神经组织浆；对照组采用单纯人羊膜管充填大鼠自体坐骨神经组织浆；标准组采用自体神经移植，桥接大鼠坐骨神经缺损。 主要观察指标：术后8，12周行大体观察、组织学检查、胫前肌湿质量、有髓神经纤维通过率及神经电生理学检测。 结果：术后12周，实验组、标准组肌萎缩有所恢复，对照组则恢复不明显。实验、标准组患侧胫前肌色泽红润，饱满富有弹性；对照组色泽相对较暗，弹性度较差。术后8，12周3组胫前肌恢复率组间比较，术后12周3组有髓神经纤维总数、截面积，神经移植体血管数和血管截面积组间比较，以及小腿三头肌复合肌动作电位幅值组间比较，差异均有显著性意义(P < 0.05），其中标准组神经纤维再生质量最佳，实验组优于对照组。 结论：肌膜转位、羊膜管预置聚乳酸-羟基乙酸微丝，并填充大鼠自体周围神经组织浆导管能较好的模拟周围神经再生之微环境，促进神经纤维再生，但与自体神经移植尚有差距。     

无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经修复坐骨神经缺损

背景：作者前期已经成功将无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经,并证明可以促进周围神经再生。 目的：构建组织工程人工神经,观察和验证桥接大鼠坐骨神经缺损后的神经功能恢复情况。 方法：成年雄性SD大鼠60只构建大鼠坐骨神经15 mm缺损模型。随机分成3组,每组20只。桥接大鼠坐骨神经缺损,实验组采用组织工程人工神经,空白对照组采用无细胞组织工程神经支架,自体神经对照组采用自体神经移植。桥接后12周通过大体观察、胫骨前肌湿质量、组织学等方法分析坐骨神经组织学及功能恢复情况。 结果与结论：桥接术后12周：实验组大鼠肢体可以支撑着地,钳夹大鼠手术侧足底皮肤出现逃避反射,足底皮肤s-100蛋白染色呈阳性反应。实验组与自体神经移植组胫骨前肌湿质量比差异无显著性意义(P > 0.05)。实验组辣根过氧化物酶逆行示踪实验显示脊髓、后根神经节均可见数量不等的辣根过氧化物酶标记阳性细胞。实验组移植物与自体神经移植组有髓神经纤维数、髓鞘厚度、神经组织面积比较差异无显著性意义。实验结果验证了无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经修复大鼠坐骨神经缺损,可以促进神经组织学的修复重建和功能的恢复。     

硫酸软骨素酶ABC(ch ABC)对大鼠坐骨神经损伤后神经再生功能的实验研究

目的观察硫酸软骨素酶ABC(chABC)对坐骨神经再生功能的影响。方法将72只SD大白鼠双侧坐骨神经切断造成0.8 cm缺损,用甲壳素导管桥接神经缺损后随机分为3组,每组24只。A组(实验组):管内注入聚乳酸-聚乙醇酸—chABC缓释微球(chABC-PLGA);B组(赋形剂组):管内注入聚乳酸-聚乙醇酸微球;C组(空白对照组):管内注入等渗盐水。术后4周、8周取材作神经电生理、神经组织学观察。结果术后4周、8周组织学观察见有再生神经通过再生室,其间有新生血管;神经电生理检查A组再生神经传导速度优于B、C组,差异有统计学意义(P<0.05),B、C组再生神经传导速度差别无统计学意义,组间比较(P>0.0167)组间多重比较行Bonferroni法检验,取校正α=0.0167)。S-100免疫组织化学及Loyez氏神经染色法显示:A组神经纤维数多于B、C组,差异有统计学意义(P<0.05),B、C组再生神经纤维数差别无统计学意义,组间比较(P>0.0167)。结论硫酸软骨素酶ABC(chABC)具有促进周围神经再生的作用。     

Tissue-engineered nerve for repair of sciatic nerve injury

Three articles regarding the use of nerve fragments bridging regeneration chambers, three-dimensional bionic nerve conduits and multiwalled carbon nanotubes for repair of sciatic nerve injury were reported in Neural Regeneration Research. We hope that our readers find these papers useful to their research.     

Sensory nerve conduction study of the mental nerve

We describe a technique for sensory nerve conduction study of the mental nerve. A monopolar recording needle is placed near the mandibular foramen using the same approach as that for routine inferior alveolar nerve block in dentistry, and a surface reference electrode is positioned over the ipsilateral mastoid process. Sensory nerve action potentials to stimulation of the mental nerve at the chin can be reliably recorded orthodromically in normal healthy subjects. The method is simple and well tolerated and provides a useful means to evaluate mental nerve function electrophysiologically.     

神经导管生物材料在神经修复中的应用

背景：神经导管是由天然或人工合成材料制成的、用于桥接神经断端的组织工程支架材料,具有引导和促进神经再生作用。 目的：总结近年来常用的神经导管生物材料在神经修复中的应用。 方法：由作者应用计算机检索维普数据库中与神经导管生物材料在神经修复中应用有关的文章,检索时限2002-01/ 2010-12。检索关键词：神经导管;生物材料;神经损伤;神经修复;神经再生。纳入标准：与神经导管生物材料在神经修复中应用有关的文章。排除标准：重复研究或较陈旧文献。根据纳入排除标准共保留相关文献30篇。 结果与结论：非生物降解材料由于其不可吸收性和对再生神经的远期不良影响使临床应用受到限制。生物降解材料在神经再生完成后可在体内降解吸收,无需二次手术取出,但目前未能利用生物降解材料完全仿制出具有天然神经结构的支架。生物衍生材料生物相容性好、排异反应小,可提供细胞外基质、胶原,起支架作用,但缺血后存在管形塌陷、再生不良、吸收瘢痕组织、增生及粘连等问题。神经导管生物材料在神经修复中的应用前景广阔,但单用一类材料难以制作出理想的神经导管生物材料,通过结合各类材料的优点,与神经营养因子、细胞外基质成分和许旺细胞等联合应用,制备新型具有生物活性的导管材料,将有利于神经修复进一步发展。     

Peripheral nerve regeneration through optic nerve grafts

Summary Grafts of optic nerve were placed end-toend with the proximal stumps of severed common peroneal nerves in inbred mice. It was found that fraying the proximal end of adult optic nerve grafts to disrupt the glia limitans increased their chances of being penetrated by regenerating peripheral nerve fibres. Suturing grafts to the proximal stump also enhanced their penetration by axons. The maximum distance to which the axons grew through the CNS tissue remained about 1.5 mm from 2–12 weeks after grafting. Schwann cells were seldom identified in the grafts. Varicose and degenerating nerve fibres were often seen within the grafts. Some varicose profiles were shown to be the terminal parts of axons within the grafts. Axons containing clusters of organelles resembling synaptic vesicles became more abundant in the longerterm grafts. Immunohistochemical studies performed on sutured grafts using a polyclonal antiserum to neurofilaments confirmed the impressions given by the electron microscopical observations. Grafts of neonatal optic nerve lacked myelin debris but were not usually penetrated by regenerating peripheral axons within a 6-week period. Sixty minutes after the intravenous injection of horseradish peroxidase, reaction product could be detected in the extracellular spaces around blood vessels in all types of living optic nerve graft. This indicates that blood-borne macromolecules could penetrate the grafts. However the profiles of axons which were found within living optic nerve grafts had no obvious relationship to blood vessels and were usually surrounded by astrocytic processes. These results suggest that living astrocytes, rather than the absence of serum-derived trophic factors or the presence of CNS myelin, constitute the major barrier to the extension of axons and the migration of Schwann cells into CNS tissue.Supported by a grant from the Wellcome Trust     

甲壳素类神经导管修复周围神经的研究与进展★

目的：概述近年来甲壳素类材料制备神经导管修复周围神经损伤的研究进展。 资料来源：应用计算机检索Medline和Springerlink 2000-01/2009-08有关神经导管材料修复周围神经损伤方面的文献，检索词“nerve conduit,peripheral nerve injury”，限定文献语言种类为“English”；同时检索中国期刊全文数据库、重庆维普数据库2000-01/2009-08相关文献，检索词“神经导管，周围神经损伤”，限定文章语言种类为中文。 资料选择：纳入与修复周围神经损伤有关的非生物降解材料、生物降解材料和生物衍生材料的文献。对资料进行初审，选取符合甲壳素类神经导管材料修复周围神经损伤要求的有关文章。排除标准相关度不大和重复性文章。 结局评价指标：神经组织工程；甲壳素类神经导管材料；神经导管制备。 结果：①随着医学材料的进展，天然或人工合成材料的神经导管用于桥接神经缺损的组织工程支架材料，具有引导和促进神经再生作用。生物可降解材料中甲壳素类神经导管可在合理的时间段内降解，有可控的生物亲和性、降解性能、多孔性和机械性能。②在导管结构、复合其他生物可降解材料、表面修饰、添加种子细胞及神经生长因子等方面进行实验研究。③通过改变再生室的空间结构和微环境，从而加快神经生长速度，促进神经功能的良好恢复。并对材料表面修饰及制备方法加以改进，使得导管适应神经再生。 结论：随着生物学技术和其他相关技术的发展，甲壳素类神经导管材料在周围神经组织工程中的应用必将得到不断的展现。 关键词：神经导管；甲壳素；生物可降解；周围神经损伤 doi:10.3969/j.issn.1673-8225.2009.47.027     

Spinal nerve composition and innervation of the axillary nerve

Introduction: The purpose of this study was to evaluate the spinal nerve composition of the axillary nerve and the contribution of each spinal nerve. Methods: Thirty brachial plexi extracted from Korean adults (15 men, 14 women, 1 unknown; left side, 13; right side, 17) were examined in this study. Results: The frequency of the contribution of C4 was 13.3%, and it was mainly innervated the deltoid and teres minor muscles. The frequency of inclusion of C7 was 20.0%. C7 fibers were mainly involved in the deltoid and cutaneous branches. The axillary nerve was primarily composed of fibers from C5 and C6 in 66.6%; C4, C5, and C6 in 13.3%; and C5, C6, and C7 in 20.0%. Conclusions: These findings may be helpful to diagnose damage to axillary nerve structures that occur in such injuries as humerus fractures. Muscle Nerve 50 : 856–858, 2014     

Determinants of nerve conduction recovery after nerve injuries: Compression duration and nerve fiber types

Introduction: The aims of this study were to determine the influences of: (1) timing of nerve decompression; and (2) nerve fiber types on the patterns of nerve conduction studies (NCS) after nerve injury. Methods: Nerve conduction studies (NCS) were performed on 3 models of nerve injury: (1) crush injury due to transient nerve compression (crush group); (2) chronic constriction injury (CCI), or permanent compression (CCI group); and (3) CCI with removal of ligatures, or delayed nerve decompression (De‐CCI group). Results: There were distinct patterns of NCS recovery. The crush and De‐CCI groups achieved similar motor nerve recovery, better than that of the CCI group. In contrast, recovery of sensory nerves was limited in the CCI and De‐CCI groups and was lower than in the crush group. Conclusions: Immediate relief of compression resulted in the best recovery of motor and sensory nerve conduction. In contrast, delayed decompression restored only motor nerve conduction. Muscle Nerve 52 : 107–112, 2015     

The role of vascularization in nerve regeneration of nerve graft

Vascularization is an important factor in nerve graft survival and function. The specific molecular regulations and patterns of angiogenesis following peripheral nerve injury are in a broad complex of pathways. This review aims to summarize current knowledge on the role of vascularization in nerve regeneration, including the key regulation molecules, and mechanisms and patterns of revascularization after nerve injury. Angiogenesis, the maturation of pre-existing vessels into new areas, is stimulated through angiogenic factors such as vascular endothelial growth factor and precedes the repair of damaged nerves. Vascular endothelial growth factor administration to nerves has demonstrated to increase revascularization after injury in basic science research. In the clinical setting, vascularized nerve grafts could be used in the reconstruction of large segmental peripheral nerve injuries. Vascularized nerve grafts are postulated to accelerate revascularization and enhance nerve regeneration by providing an optimal nutritional environment, especially in scarred beds, and decrease fibroblast infiltration. This could improve functional recovery after nerve grafting, however, conclusive evidence of the superiority of vascularized nerve grafts is lacking in human studies. A well-designed randomized controlled trial comparing vascularized nerve grafts to non-vascularized nerve grafts involving patients with similar injuries, nerve graft repair and follow-up times is necessary to demonstrate the efficacy of vascularized nerve grafts. Due to technical challenges, composite transfer of a nerve graft along with its adipose tissue has been proposed to provide a healthy tissue bed. Basic science research has shown that a vascularized fascial flap containing adipose tissue and a vascular bundle improves revascularization through excreted angiogenic factors, provided by the stem cells in the adipose tissue as well as by the blood supply and environmental support. While it was previously believed that revascularization occurred from both nerve ends, recent studies propose that revascularization occurs primarily from the proximal nerve coaptation. Fascial flaps or vascularized nerve grafts have limited applicability and future directions could lead towards off-the-shelf alternatives to autografting, such as biodegradable nerve scaffolds which include capillary-like networks to enable vascularization and avoid graft necrosis and ischemia.     

Hemihypoglossal-facial nerve anastomosis for facial nerve palsy

BACKGROUND AND PURPOSE : Commonly used classic hypoglossal (CN XII) to facial nerve (CN VII) anastomosis has the disadvantage of tongue hemiatrophy. Thus, various attempts have been made to modify this method to reduce the tongue damage. The aim of this report was to present the results of hemihypoglossal-facial nerve anastomosis (HHFA) technique in relation to facial muscles reanimation and hemitongue atrophy. MATERIAL AND METHODS : The first 7 consecutive patients who underwent CN VII anastomosis with half of the CNXII, for which the follow-up period exceeded 12 months, were analysed. During the procedure, CN VII was transected as proximally as possible after drilling the mastoid process. CN XII was separated longitudinally into two parts at a short distance to allow suture of the stumps without any tension. One half of CN XII was transected and sutured to the distal stump of CN VII. Recovery from facial palsy was quantified with the House-Brackmann grading system (HB). Tongue function was assessed according to the scale proposed by Martins. RESULTS : Features of initial reinnervation of facial muscles were visible after 6 months in all 7 patients. All patients achieved satisfactory outcome of CN VII regeneration (HB grade III) until the last control examination (12-27 months after surgery, mean 16). No or minimal tongue atrophy without deviation (grades I-II according to the Martins scale) was found in 4 patients. Mild hemiatrophy with tongue deviation < 30 degrees (grade III) was visible in 3 patients. CONCLUSIONS : In our experience, HHFA is effective treatment of facial palsy and gives a chance to reduce damage of the tongue.