丙戊酸钠充填导管促进大鼠外周神经再生的实验研究

目的 观察丙戊酸钠(VPA)充填导管对缺损外周神经再生的促进作用.方法 通过建立大鼠坐骨神经缺损模型.用硅胶管(1 cm)进行缺损神经段(0.8 cm)的桥接,局部应用8%VPA注射液10ul,观察VPA对神经再生和运动神经功能恢复的影响.30只大鼠随机分成2组,实验组在硅胶管局部注射VPA注射液;对照组在硅胶管局部注入生理盐水. 结果 术后每只大鼠每2周进行坐骨神经功能指数(SFI)检测,每4周做电生理检查,最后术后12周处死所有大鼠,对坐骨神经进行组织形态学分析.用数字图像分析软件检测有髓神经纤维髓鞘厚度.并对再生神经纤维轴突计数.通过统计软件分析发现SFI,电生理检查,神经组织性形态上两组差异均有统计学意义(P＜0.05). 结论 局部使用VPA于神经缺损的大鼠,可以促进损伤神经的轴突再生和运动功能恢复.因此VPA有望在临床上应用于外周神经损伤病例的治疗.     

Enhanced rat sciatic nerve regeneration through silicon tubes implanted with valproic acid

Valproic acid (VPA) is an effective antiepileptic drug and mood stabilizer. It has recently been demonstrated that VPA could promote neurite outgrowth, activate the extracellular signal-regulated kinase pathway, and increase B-cell lymphoma/leukemia-2 (bcl-2)and growth cone-associated protein 43 (GAP-43) levels in spinal cord. We hypothesized that VPA could enhance axonal regeneration in the rat. In the present research, we demonstrate the effect of VPA on peripheral nerve regeneration and recovery of motor function through a silicon tube implanted with VPA. The left sciatic nerves were exposed through dorsal-splitting incisions, and 8-mm nerve sections were excised at the middle of the thigh. Then, a 1.0-cm-long silicone tube (internal diameter,1.0 mm; exterior diameter, 2.0 mm) was used to bridge the nerve deficit, anchored to the proximal and distal terminals of the excised deficit of sciatic nerves with 9-0 nylon epineural suture. Sterile petroleum jelly was used to seal the ends of the tubes to avoid leakage. The rats in the VPA group and control group were locally delivered 10 muL VPA injection (400 mg/5 mL) and normal saline, respectively, after the operation. The sciatic nerve index (SFI) was observed in each animal at 2-week intervals and electrophysiology was studied at 4-week intervals for 12 weeks. Histological and morphometrical analyses were performed at the end of the experiment (12 weeks after the operation). Using the digital image-analysis system, the thickness of the myelin sheath was measured, and total numbers of regenerated axons were counted. There was a significant difference in SFI, electrophysiological index (motor-nerve conduct velocity, amplitude of activity potential), and morphometrical results (regenerated axon number and thickness of myelin sheath) in nerve regeneration between the VPA group and controls ( P < 0.05). The results demonstrated that VPA is able to enhance sciatic nerve regeneration in rats, suggesting the potential clinical application of VPA for the treatment of peripheral nerve injury in humans.     

细胞外ATP对外周神经再生作用的实验研究

目的：研究细胞外ATP对外周神经再生的影响。方法：采用大鼠坐骨神经硅胶管再生室模型,左侧再生室中加入1mmol/LATP作为实验组,右侧再生室中加入生理盐水对照。术后1、2、4、6和12周取材,每次取材除作肉眼观察外,第6、12地行光镜及电镜观察,并采用图像分析检测再生轴突数目和髓鞘厚度。结果：ATP实验组再生神经干的直径,神经干内有髓纤维的数目以及髓鞘的厚度同对照组相比 ,差异非常显著。电镜观察表明,实验组再生随鞘的厚度及成熟情况明显优于对照组。结论：细胞外ATP具有较强的促进神经再生的作用。     

含神经生长因子的几丁质管桥接兔面神经缺损的实验研究

目的 探讨含神经生长因子（ＮＧＦ）的几丁质管在修复兔面神经缺损中的作用。方法 将１６只新西兰兔的两侧面神经上颊支分别造成８ｍｍ的缺损,左侧用管腔内注入ＮＧＦ的几丁质管修复,右侧用自体神经移植修复作对照。术后８、１６周,分别取８只兔进行电生理和超微结构研究。结果 实验侧术后８周,再生神经中有髓和无髓神经纤维排列整齐,有髓神经的髓鞘厚,板层结构清晰;术后１６周,再生神经中有髓和无髓神经纤维数量增加,形     

联合使用ATP和NGF对周围神经再生作用的实验研究

目的 研究三磷酸腺苷(adenosine tri-phosphate，ATP)和神经生长因子(nerve growth factor，NGF)联合使用时对受损周围神经再生的作用并比较两者作用的强弱。方法 将64只SD大鼠左侧坐骨神经切断后直接作端端缝合，缝合段置于硅胶管室内。根据室内注入药物的不同，分为生理盐水(NS)、ATP、NGF、和ATP加NGF4组，每组16只大鼠。术后4周和8周(每组均为8只)取标本，作形态学，肌湿重，运动神经传导速度(MNCV)、复合运动动作电位(CMAP)波幅，及组织学的检测。结果 ATP加NGF组的神经纤维数目、大小、髓鞘厚度和MNCV、CMAP及肌湿重均优于其它3个组。NGF组的各项指标比ATP组好。结论联合使用ATP和NGF时受损周围神经的再生作用明显增强。NGF的作用强于ATP。     

靶肌肉注射睫状神经营养因子促周围神经再生的功能评价

目的 评价靶肌肉注射睫状神经营养因子(Ciliary Neumtrophic Factor，CNTE)对受损周围神经功能恢复的影响。方法 用硅胶管桥接80支大鼠左侧6mm长坐骨神经缺损，随机分为2组。分别行CNTF、生理盐水(NS)靶肌肉注射。术后行坐骨神经功能指数(SFI)测定、电生理检测、轴突图像分析及霍乱毒素-辣根过氧化物酶(CB-HRP)逆行追踪。结果 CNTE组SFI恢复率、各项电生理及轴突图像分析指标、CB-HRP标记的脊髓前角运动细胞数明显优于NS组。结论 靶肌肉注射CNTF可明显促进周围神经再生，提高神经功能恢复。     

Siatic nerve regeneration in rats stimulated by fibrin glue containing nerve growth factor: an experimental study

Objective

To study the effects of fibrin sealant containing nerve growth factor on the peripheral nerve regeneration.

Study design

A four-group experimental design with repeated measures on one factor was used.

Methods

Fibrin glue (FG) containing NGF was injected into the site of end-and-end sutured peripheral nerve (sciatic nerve) (group I: NGF + FG), meanwhile three control groups were set-up: group II (NGF), group III (FG), and group IV (normal saline). Observation to the function and morphology of the sciatic nerve was carried out at the end of 4, 8, 12 weeks postoperation. Data were analyzed using ANOVA, with the appropriate post hoc between-groups comparison test.

Results

Electrophysiological testing. The NAP and NCV of group I (NGF + FG) were greater than those of group II (NGF), group III (FG), or group IV (normal saline) (p < 0.05). Sciatic functional index (SFI). It began to ameliorate at 4 weeks postoperation and SFI increased as time went on. And the SFI in group I (NGF + FG) was better than those in group II (NGF), group III (FG), or group IV (normal saline) (p < 0.05). Morphology. In the MGF-stained sections, dissociated myelin debris was less and regenerated nerve fibres were in larger quantities in group I (NGF + FG) than in other groups. In the HE-stained sections, regenerated nerve fibres distal to anastomosis significantly increased, and axon and myelin had a clearer outline in group I (NGF + FG) than in other groups. Electron microscopy indicated that the regenerated nerve fibres were more mature and the development of the axons was greater in group I than in other control groups.

Conclusions

FG can be used as carrier of exogenous NGF, and they can provide synergistic effects for the peripheral nerve regeneration after the integration of the two.     

Nerve growth factor: increased angiogenesis without improved nerve regeneration.

Nerve growth factor (NGF) and laminin are important factors for neural development and regeneration. We examined the effects of increasing the local concentration and duration of action of NGF and laminin on peripheral nerve regeneration in the adult mouse sciatic nerve. A Silastic (silicone rubber) channel with intraluminal NGF solution was secured between transected nerve ends. The second channel tested, formed from a polysaccharide called chitosan, was prepared with NGF and laminin in the channel walls and provided a sustained release of NGF. At six weeks post-implantation, no improvement in nerve regeneration was identified in those channels prepared with NGF when comparing electromyographic thresholds (microA), maximum potentials (mV), nerve diameter, myelin sheath thickness, myelinated axon counts, or diameter. However, increased angiogenesis was demonstrated within the chitosan and Silastic channels prepared with NGF compared to those channels without NGF. Silastic exhibited minimal inflammation. Chitosan was associated with inflammation in many nerve channels.     

促红细胞生成素促进大鼠坐骨神经再生作用的实验研究

目的 探讨促红细胞生成素(Erythoropoietin,EPO)对大鼠坐骨神经损伤修复后神经再生的影响,为周围神经损伤的临床治疗提供实验依据.方法 雄性SD大鼠40只,随机分为两组,即EPO组和神经生长因子(NGF)组,用硅胶管桥接10 mm的坐骨神经缺损,EPO组和NGF组分别注射EPO和NGF.术后4周和8周时每组分别提取10只大鼠,以坐骨神经功能指数(SFI)、运动神经传导速度(MNCV)、形态学观察和蛋白基因产物9.5(PGP 9.5)免疫组织化学染色,评估EPO对大鼠坐骨神经再生的影响.结果 术后4周SFI,EPO组为[(-78.85±3.87),x-±s,下同],NGF组为(-79.98±4.58),差异无统计学意义(P>0.05);术后8周SFI,EPO组为(-60.26±2.91),NGF组为(-64.65±4.11),差异有统计学意义(P<0.05).术后4周和8周时,EPO组MNCV、有髓神经纤维数目以及PGP9.5免疫阳性神经纤维的平均光密度和积分光密度均优于NGF组,差异有统计学意义(P<0.05).结论 EPO 能促进大鼠坐骨神经损伤后的修复与再生.     

靶肌肉注射睫状神经营养因子促周围神经再生的功能评价

目的　评价靶肌肉注射睫状神经营养因子 (CNTF)对受损周围神经功能恢复的影响。方法　用硅胶管桥接 80只大鼠左侧坐骨神经长 6 m m缺损 ,随机分为两组 ,分别行 CNTF、生理盐水 (NS)靶肌肉注射。术后行坐骨神经功能指数 (SFI)测定、电生理检测、轴突图像分析及霍乱毒素 -辣根过氧化物酶 (CB- HRP)逆行追踪。结果　 CNTF组 SFI恢复率、各项电生理及轴突图像分析指标、CB- HRP标记的脊髓前角运动细胞数明显优于 NS组。结论　靶肌肉注射 CNTF可明显促进周围神经再生和神经功能恢复。     

新型纤维素导管桥接周围神经缺损的研究

目的运用一种新型纤维素材料导管桥接周围神经缺损。方法将原代培养、纯化的Schwann细胞种植入新型复合纤维素导管作为移植物，于大鼠模型上桥接1．0cm坐骨神经缺损。术后1周，扫描电镜观察Schwann细胞在导管内壁的生长情况；术后12周，取桥接物中段,行半薄切片光镜观察及超薄切片电镜观察神经纤维再生状况。结果桥接物植入后1周,Schwann细胞贴附于导管内壁良好生长。植入后12周，管内有大量排列整齐、结构成熟的再生神经纤维。结论新型纤维素组织工程化神经可有效桥接周围神经缺损，该导管材料在医疗中具有广阔的应用前景。     

不同时段异体神经片段皮下包埋对周围神经再生影响的实验研究

目的 探讨异体神经片段经皮下包埋不同时段后对周围神经再生的影响。方法 Wistar大鼠55只，雌雄不限，随机分为5组，A、B、C组（实验组）和D组（对照组）每组各10只，E组（供体组）15只。E组动物在出骨盆口以远5mm处切断双侧坐骨神经，向远端游离约15mm，切断作为移植物。A、B、C组动物均行左侧大腿切口，皮下钝性分离，埋入供体神经片段。术后1周（A组）、2周（B组）、3周（C组）显露右侧坐骨神经，距骨盆出口约5mm处切断，向远端游离约10mm再切断，取出对侧包埋的神经片段，修剪远近端保留长度约10mm，移植于右侧神经缺损处。D组显露右侧坐骨神经后，在距骨盆出口约5mm处切断，向远端游离约10mm后再切断，原位缝合。术后2、4、6、8、10及12周监测坐骨神经功能指数（sciatic functional index，SFI），术后12周行电生理检查测试运动神经诱发电位的传导速度及潜伏期，组织学检测移植神经再生轴突数目和面积，以及移植神经的超微结构变化。结果术后各组SFI逐渐下降，12周时A组和D组的SFI最小，两组间差异无统计学意义，但分别与B组和C组比较差异有统计学意义（P〈0．05）。术后12周A组和D组再生大量有髓神经纤维及少量无髓神经纤维，再生神经的数量和结构与正常神经相似，图像分析显示两组间无明显差别，与B组和C组比较差异有统计学意义（P〈0．05）。A组和D组的运动神经传导速度及潜伏期结果无差异，优于B组和C组，且差异有统计学意义（P〈0．05）。结论 异体神经片断皮下包埋后有促进周围神经再生的作用，皮下包埋1周组促神经再生作用优于皮下包埋2、3周组。     

睫状神经营养因子神经生长因子对周围神经再生的作用

目的：研究睫状神经营养因子（ｃｉｌｉａｒｙｎｅｕｒｏｔｒｏｐｈｉｃｆａｃｔｏｒ，ＣＮＴＦ）和神经生长因子（ｎｅｒｖｅｇｒｏｗｔｈｆａｃｔｏｒ，ＮＧＦ）对周围神经再生的作用。方法：硅管套接大鼠坐骨神经，将ＣＮＴＦ、ＮＧＦ、生理盐水（ｎｏｒｍａｌｓａｌｉｎｅｓｏｌｕｔｉｏｎ，ＮＳ）分别加入硅管中，并于术后每日皮下注射一定剂量。术后４周，应用ＨＲＰ逆行追踪技术显示再生轴突通过修复部位的胞体。结果：与ＮＳ组、ＮＧＦ组相比，ＣＮＴＦ组脊髓标记胞体显著增加，其它组间比较，差异不显著。结论：外源性ＣＮＴＦ能明显促进周围神经运动轴突再生，但促进感觉轴突再生的作用不明显。外源性ＮＧＦ促进周围神经运动和感觉轴突的再生均不明显     

Pyrroloquinoline quinone enhances regeneration of transected sciatic nerve in rats

Slowregenerationofinjuredperipheralnerve,asamajorreason,contributesalottothepooroutcomeofneuralfunctionalrecovery.Satisfactorynerverehabilitationnotonlydependson applyingaperfectskillofmicrosurgery,butalsoneeds afitenvironmentfornerveregeneration.Drugsfor…     

Enhancement of motor nerve regeneration by nerve growth factor.

The sciatic nerves of adult Wistar rats were severed bilaterally. Each nerve was sutured into a silicone tube used as a conduit, leaving a 5 mm gap in length between the nerve ends. Nerve growth factor in a saline solution vehicle was injected into the silicone chamber on the right side and normal saline solution (control) on the left. Six weeks after surgery, electrophysiological studies were performed. The motor nerve conduction velocities (MNCV) were significantly increased in the NGF-treated nerves. In one rat, the MNCV on the NGF-treated side was 66.6 m/s, in the range of normal nerves. There was no significant difference between the two sides in the amplitudes of evoked muscle action potentials. There are apparently no reports on the effect of NGF on motor neuron regeneration in vitro. In this study, NGF was found to enhance motor nerve regeneration.     

外消旋聚乳酸/神经生长因子复合膜包绕神经缝接部位促进神经再生

目的探讨外消旋聚乳酸／神经生长因子（PDLLA／NGF）复合膜包绕神经缝接部位是否具有促进神经再生作用。方法雌性Wistar大白鼠16只，随机分为2组：A组（神经端端缝接）8只；B组（神经端端缝接并缝接部位包绕PDLLA／NGF复合膜）8只。显露每只动物的右侧坐骨神经，利刀横断。A组以外膜缝合法端端缝接；B组以外膜缝合法端端缝接后，在缝接部位包绕1层PDLLA刷GF（含量250U）复合膜。6个月后，观察比较两组神经再生情况。结果B组的小腿三头肌湿重恢复率、运动神经传导速度、再生的有髓神经纤维直径、轴突直径、髓鞘厚度均优于A组。结论PDLLA／NGF复合膜包绕神经缝接部位具有促进神经再生作用。     

微囊化转NGF基因3T3细胞修复大鼠周围神经损伤的实验研究

目的 探讨微囊化转NGF基因3T3细胞移植促进神经再生的可行性。方法 带有小鼠NGF基因的质粒pcDNA3．1 ／NGF经FuGENE^TM6转染NIH 3T3细胞，用APA(海藻酸钠-多聚赖氨酸-海藻酸钠)微胶囊包埋后，进行体外培养，定期检测微囊化细胞活性和NGF分泌量变化。同时，将转基因细胞移植于大鼠坐骨神经切断吻合处，于手术后4、8、12周检测神经再生和功能恢复结果。结果 微囊化转NGF基因3T3细胞在体外培养条件下可存活3个月并能稳定表达，移植于损伤坐骨神经的微囊化转NGF基因细胞组再生神经密度大，排列有序，吻合口瘢痕少，单核细胞浸润少，水肿轻，动作电位幅值和传导速度显著增大，坐骨神经功能指数明显升高。结论 微囊化转NGF基因3T3细胞移植可显著促进神经再生和降低异体细胞移植免疫反应。     

胶质细胞源性神经营养因子基因修饰的雪旺氏细胞修复大鼠周围神经缺损的实验研究

目的　从转基因角度探讨治疗周围神经损伤的有效方法。方法　成年Wister大鼠 4 8只 ,平均分为 3组。切断大鼠坐骨神经并形成 10mm长缺损 ,用硅胶管桥接两侧断端 ,管腔内植入胶质细胞源性神经营养因子 (glialcell linederivedneurotrophicfactor,GDNF)修饰的雪旺氏细胞 (schwanncells,SCs) ,正常SCs修复组和单纯硅胶管修复组作为对照 ,分别于术后 4、8、12和 16周对各组动物进行大体观察 ,肌电图测量 ,组织学切片观察 ,再生神经的神经电生理检测 ,GDNF免疫组化检测 ,组织学切片 ,观察和图像分析。结果　GDNF SCs组动物的神经传导速度、有髓神经纤维密度、神经组织面积、髓鞘厚度均显著优于SCs组和硅胶管组。结论　将GDNF基因修饰的雪旺氏细胞移植修复周围神经缺损 ,使局部释放的GDNF维持神经元存活 ,加快轴突再生速度以促进周围神经再生 ,此方法为将来治疗周围神经损伤提供了线索。     

The epineural sleeve technique for nerve graft reconstruction enhances nerve recovery

The purpose of the study was evaluation of nerve recovery following epineural sleeve technique for graft reconstruction in rat sciatic nerve. This technique provides the epineural sleeve to cover and separate the site of coaptation. Animals were divided into three groups: CNG-conventional nerve grafting, ESN-epineural sleeve from recipient nerve stumps, ESG-epineural sleeve from graft. Nerve regeneration was evaluated by pin-prick, toe-spread test, walking track analysis and somatosensory-evoked potentials (SEP), gastrocnemius index (GI), and histomorphometric evaluation. Most parameters (SFI, SEPs, and GI) showed significantly better nerve recovery for ESN group when compared to conventional CNG group. Also ESG group revealed better result for SFI. Better functional results for ESN and ESG groups were further confirmed by histomorphometric analysis: higher axon density and diameters as well as thicker myelin sheath. Epineural sleeve graft technique may be promising method with potential application for nerve reconstructive procedures. Better functional nerve recovery can be anticipated.     

静电纺纳米聚乳酸己内酮共聚物导管修复周围神经缺损的实验研究

目的 探讨应用同轴静电纺丝技术制备的聚乳酸己内酮共聚物[Poly(1-lactide-co-epsilon-caprolactone),P(LLA-CL)]导管,移植修复大鼠周围神经缺损的效果.方法 选取健康SD大鼠54只,随机分成3组,每组18只.先造成坐骨神经1.5cm缺损段,然后分别采用P(LLA-CL)导管桥接(A组)、硅胶管桥接(B组)、自体神经逆行原位移植(C组).分别在术后4、8、12周对大鼠进行大体观察、坐骨神经功能指数检查、神经电生理检查、肌肉湿重、再生有髓神经纤维计数、电镜观察,评价各组神经再生.结果 术后4周时A组再生神经已部分生长到导管的中部;8周时再生神经已通过神经导管,但再生的神经纤细;12周时再生神经粘连较轻,直径较粗.A组的坐骨神经功能指数、神经电生理、肌肉湿重和组织学观察等各项指标均略差于C组,但明显优于B组.结论 纳米聚乳酸己内酮神经导管具有促进神经轴突再生的作用,有望成为自体神经移植的替代材料应用于周围神经缺损的修复.