周围神经端侧吻合后神经再生的研究

目的研究周围神经端侧吻合后远端神经的再生及其临床应用价值。方法选用３７只Ｗｉｓｔａｒ大鼠，分成Ａ、Ｂ两组。Ａ组：切断腓神经，在邻近的胫神经干外膜上开一１ｍｍ小窗，将腓神经远端吻合到胫神经干侧方开窗处。Ｂ组：切断腓神经后，在胫神经干和远侧腓神经干外膜上各开一小窗，取对侧相应的腓神经以端侧吻合的方式桥接于胫、腓神经干两窗之间。两组分别于术后４、８、１２周取材，作神经组织学、电镜及图像分析仪检测：（１）神经再生的数量和质量；（２）定性定量观察神经再生情况；（３）再生神经的数目、密度及截面积。结果直接端侧吻合或神经桥接移植的端侧吻合，均可使神经再生，且再生纤维的质量与端端吻合组（对照组）无显著性差异（Ｐ＞０．０５）；对被吻合神经－胫神经的功能亦无影响。结论神经端侧吻合可作为神经损伤后一种新的修复方法     

Comparative electrophysiologic evaluation of nerve grafts and autogenous vein grafts as nerve conduits: an experimental study

This study was carried out to compare electrophysiologically the efficacy of autogenous vein grafts, with autogenous nerve grafts as conduits for nerve regeneration. A 0.75-cm segment of sciatic nerve was resected in two groups of Sprague-Dawley rats of equivalent maturity. The nerve gaps were bridged with an autogenous vein graft in the first group (31 rats), and an autogenous nerve graft in the second group (24 rats). Serial in vivo nerve conduction velocity studies and terminal in vitro nerve conduction velocity and nerve action potential measurements were performed. An additional group of 21 animals who had undergone no surgical procedures, were similarly studied to establish an age-adjusted baseline for comparison. Twelve animals in the first group, 14 in the second group, and 13 in the baseline group survived the full year of study. In vivo conduction velocities between the two experimental groups compared favorably. Nerve conduction velocity determined by in vitro technique confirmed this finding and measured similarly at about 78 percent of the baseline. Nerve action potential amplitude in the vein-grafted group was 12.0 percent of the baseline, while the nerve-grafted group was 23.9 percent of the baseline. This study demonstrated that the vein graft compares well with the nerve graft in nerve conduction velocity, but only one-half as well in nerve action potential.     

大鼠股神经再生趋化性效果评价

[目的]评价股神经损伤后神经功能的恢复效果,进一步探讨股神经趋化性再生的评价方法。[方法]雄性Wistar大鼠30只,分为对照组、电生理检测组、标记组,每组10只,于右侧股神经分叉上4 mm处行断端吻合术。术后8周,行电生理和组织学检查,并分别用F488和DiI逆行示踪标隐神经和股神经肌支,观测脊髓前角中示踪剂的分布及数目。[结果]实验动物术侧活动受限;术侧运动诱发电位潜伏期:2.16 ms±0.44 ms,波幅:2.01 mv±0.70 mv,与正常侧相比,其潜伏期延长(P<0.05),波幅降低(P<0.05);NF-200染色显示神经断端吻合口连续性良好;标记组脊髓前角中被标记的神经元颜色和数目分别为绿色:80.2±9.6、红色:200.6±17.9、橙色:26.9±4.4,而对照组中仅为红色:726.7±119.0(P<0.05)。[结论]在进行周围神经损伤再生评价时,除了行为学、电生理和组织形态学常规检测外,采用逆行示踪标记神经元计数的方法可以作为趋化性再生效果评价的一个选择。     

电针对大鼠坐骨神经再生影响的功能评价

目的从功能恢复评价电针对神经再生的促进作用。方法建立大鼠坐骨神经损伤模型，将９０只ＳＤ大鼠随机分为损伤对照组和电针组两组。分别于术后２、４、６、１０周测定运动神经传导速度、小腿三头肌肌力及坐骨神经功能指数，并以自身健侧作对照得出其恢复率。同时测定术后１周感觉神经再生的距离。结果电针组术后１周感觉神经再生距离及２、４、６、１０周运动神经传导速度、肌力、ＳＦＩ等的恢复率均明显优于对照组。结论电针可以促进损伤神经的再生，明显提高神经功能的恢复     

Fiber regeneration in nerve grafts without connection to a target muscle: An experimental study in rabbits

In 30 rabbits, both saphenous nerves were harvested as autografts and coapted to the branch for the rectus femoris muscle without connection to any distal target muscle. The graft from the right thigh was led to the contralateral extremity (crossover grafting). The graft on the left side remained on the same extremity (ipsilateral grafting). Animals were separated into four groups and were sacrificed 3, 6, 9, and 12 months after grafting. Specimens of the grafts and the donor motor branches were harvested for histomorphometric examination. The number of myelinated nerve fibers in the distal end of the nerve grafts was significantly increasing from 3 to 6 months after grafting and remained on a relatively constant level in the long-term groups. Three months after the first operation, greater numbers of myelinated nerve fibers were counted after ipsilateral grafting than after crossover grafting. In the long-term groups, this difference could not be observed. © 1993 Wiley-Liss Inc.     

不同端侧缝合方法对周围神经再生的影响

目的比较各种不同的端侧缝合方法治疗周围神经损伤后神经再生的优劣。方法ＳＤ大鼠９０只，随机分成５组。前４组右侧腓总神经作端侧缝合，左侧切除长１．５ｃｍ的腓总神经作为对照。Ａ组：右侧远断端与近段行侧端吻合，外膜不开窗。Ｂ组：方法同Ａ组，但束、外膜开窗。Ｃ组：右侧近断端与远段行端侧吻合，外膜不开窗。Ｄ组：方法同Ｃ组，但束、外膜开窗。Ｅ组：右侧以４５度角、左侧以９０度角作侧端缝合。各组分别于术后１、２、３个月取材，作肌电图、组织学及肌湿重检查。结果Ａ与Ｂ，Ｃ与Ｄ组比较，运动神经诱发电位潜伏期、波幅及有髓纤维计数，后者优于前者（Ｐ＜０．０５）。肌湿重及肌纤维截面积，两者无明显差异（Ｐ＞０．０５）。４５度与９０度角比较，前者神经再生质量明显优于后者（Ｐ＜０．０５）。结论周围神经可通过端侧吻合而再生，以束、外膜开窗及４５度角的缝合方法为佳     

Nerve repair: experimental and clinical evaluation of neurotrophic factors in peripheral nerve regeneration

SUMMARY: Neurotrophic factors are a family of polypeptides required for survival of discrete neuronal populations. In the normal state such factors are mostly synthesised by target tissues and are used for the viability of the nerve-cell bodies. After nerve injury, neurotrophic factors (NFs) are synthesised by non-neuronal (Schwann cells and fibroblasts) in the nerve trunk, and act to support the outgrowth of axons. NFs can be classified into three major groups: (1) neurotrophins; (2) neurokines; and (3) the transforming growth factor beta (TGF)-beta superfamily.     

膈神经移位术后神经元再生的实验研究

目的 通过观察膈神经移位术后大鼠神经元再生情况及其形态学改变,了解该神经移位术后运动和感觉神经元的再生能力.方法 采用SD成年雌鼠(10～12周龄,n=11只),一组大鼠膈神经切断后用荧光示踪剂快蓝溶液标记,一周后经心脏灌注4%副甲醛溶液,取出C2-5脊髓节段及背根神经节,切片在荧光显微镜放大250倍下计数快蓝标记的运动神经元和感觉神经元,并测量荧光标记的神经元截面积.另一组大鼠行膈神经移位至肌皮神经,经过6个月的神经再生后,在吻合口远端切断肌皮神经,采用同样方法逆向标记已经完成再生的神经元,一周后如上法取材,计数神经元及测量胞体截面积.结果 1.正常大鼠膈神经核含运动神经元[(344.3±10.0)个,(x)±s,下同],截面积为(588.5±31.9)μm2;含感觉神经元(427.0±54.6)个,截面积为(881.9±86.9)μm2.2.膈神经移位术后,约有95%的运动神经元(326.0±16.3)个完成了再生,再生的运动神经元胞体表现出轻度肿胀(673.6±25.8)μm2;只有约60%的感觉神经元完成了再生(255.8±45.2)个,再生的感觉神经元表现出胞体萎缩(668.8±51.1)μm2.结论 膈神经移位后具有良好的神经元再生能力,尤其表现在运动神经元,为术后运动功能恢复创造了重要的先决条件.     

Axon regeneration and vascularisation of nerve grafts an experimental study

An experimental model has been designed to assess the effect of vascularisation on axon regeneration in nerve grafts. The vascular status of the grafts has been demonstrated by microangiography and histology. Rat sciatic nerve grafts in which the vascular pedicles were left intact retained a normal vascular pattern which was not adversely affected by wrapping the graft in a polythene sleeve. In devascularised grafts, revascularisation commenced at three days and was complete at nine days. If the devascularised grafts were wrapped in a polythene sleeve, revascularisation was impeded and at fifteen days the middle segment of the graft was avascular and infarcted. The rate of axon regeneration was measured electrophysiologically in the above four groups of nerve grafts. There was a linear relationship between the rate of axon regeneration with time post-graft, axon growth proceeding at a mean rate of 1.150mm/day (S.E. +/- 0.084) after a mean delay of 4.85 days. There was no significant difference in the rate of axon regeneration in the four groups.     

Experimental study of vascularized nerve grafts: morphometric study of axonal regeneration of nerves transplanted into silicone tubes

Rat sciatic nerves were used in a comparative study of vascularized and free (nonvascularized) nerve grafts transplanted into silicone tubes. A total of 39 sciatic nerves were used, 21 as vascularized nerve grafts and 18 as free nerve grafts. The rats were killed at intervals from the first to the 24th postoperative weeks, and biopsied sciatic nerves were processed for morphometric studies using a fiber caliber analyzer. There was a significant increase in the number of large myelinated axons (more than 5 mu) in the vascularized nerve graft models over those in the free nerve graft models at the distal ankle region. Furthermore, the diameters of myelinated axons in the vascularized nerve graft were larger than those in the free nerve graft at all specimen sites during all postoperative weeks. We suggest that the preservation of the vascular system in vascularized nerve grafts would decrease the likelihood of fibrosis and result in better regeneration of axons.     

Experimental study of vascularized nerve graft: evaluation of nerve regeneration using choline acetyltransferase activity

A comparative study of nerve regeneration was performed on vascularized nerve graft (VNG) and free nerve graft (FNG) in Fischer strain rats. A segment of the sciatic nerve with vascular pedicle of the femoral artery and vein was harvested from syngeneic donor rat for the VNG group and the sciatic nerve in the same length without vascular pedicle was harvested for the FNG group. They were transplanted to a nerve defect in the sciatic nerve of syngeneic recipient rats. At 2, 4, 6, 8, 12, 16, and 24 weeks after operation, the sciatic nerves were biopsied and processed for evaluation of choline acetyltransferase (CAT) activity, histological studies, and measurement of wet weight of the muscle innervated by the sciatic nerve. Electrophysiological evaluation of the grafted nerve was also performed before sacrifice. The average CAT activity in the distal to the distal suture site was 383 cpm in VNG and 361 cpm in FNG at 2 weeks; 6,189 cpm in VNG and 2,264 cpm in FNG at 4 weeks; and 11,299 cpm in VNG and 9,424 cpm in FNG at 6 weeks postoperatively. The value of the VNG group was statistically higher than that of the FNG group at 4 weeks postoperatively. Electrophysiological and histological findings also suggested that nerve regeneration in the VNG group was superior to that in the FNG group during the same period. However, there was no significant difference between the two groups after 6 weeks postoperatively in any of the evaluations. The CAT measurement was useful in the experiments, because it was highly sensitive and reproducible.     

An experimental study of nerve regeneration through chemically treated allografts

Summary We carried out experiments in rabbits to determine whether treating nerve transplants with gradually increasing concentrations of ethanol, ether and ficin would inhibit the graft-host immune reaction to the allograft. After treatment with ethanol, the basal laminar scaffold of the Schwann cell remained intact and there was satisfactory axonal regeneration. The results after additional treatment with ether or ficin did not achieve such good results. Preservation of the basal lamina is considered to be the essential factor in allowing neural regeneration in these circumstances.

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Résumé Nous avons réalisé une expérimentation sur le lapin afin de déterminer s'il était possible de supprimer chez le receveur les réactions immunitaires aux allogreffes, ceci en traitant les transplantations nerveuses à l'aide de concentrations progresivement croissantes d'éthanol, d'éther et de ficine. Après traitement par l'éthanol l'échafaudage laminaire basal des gaines de Schwann reste intact et il se produit une repousse correcte des axones. Les résultats ne sont pas aussi bons après traitement par l'éther ou par la ficine. La préservation de la laminaire basale est donc le facteur essentiel qui permet, dans cette situation, la régénération nerveuse.

     

外源性表皮生长因子促进鼠坐骨神经再生的实验研究

目的 评价外源性表皮生长因子（exogenous epidemal growth factor,EGF）对神经再生的影响。方法 雄性SD大鼠48只,建立成鼠坐骨神经挤压伤模型。按术后注射药物的不同成分2组,每组24只鼠。损伤对照组：在神经损伤处注射生理盐水5μl;EGF组：注射EGF／生理盐水液（10μg/5μl）。于术后2、4、6周3个时间点测定坐骨神经功能指数、CMAP的潜伏期、最大语诱发电位的恢复率、组织学检测、电镜超微结构观察。结果 坐骨神经功能指数恢复率在各时间点,EGF组无明显优于对照组（P＜0．01）。CMAP潜伏期的延迟率,EGF组明显小于对照组（P＜0．01）;诱发电位恢复率EGF组明显好于对照组（P＜0．01）。组织学检查：有髓神经纤维数在术后2、4周时EGF组明显多于对照组（P＜0．01）;各时间点有髓神经纤维直径及截面积,EGF组明显优于对照组（P＜0．01）。超微结构观察：EGF组再生神经的有髓纤维数,髓鞘厚度,髓鞘的成熟度明显好于对照组。结论 外源性EGF对神经的再生和功能恢复有一定的作用。     

周围神经侧侧缝合法的实验研究

目的 提出一种修复周围神经操作的新方法－－侧侧缝合法,并对侧侧缝合后神经的再生模式进行初步研究。方法 选用ＳＤ雄性大鼠１２只,双下肢随机分为实验侧和对照侧。实验侧：将腓总神经在大腿下１／３处切断,断端结扎后将其远端与相邻胫神经干适当松解后靠拢,纵行切开两神经相邻侧面的神经外膜、束膜长约０．５ｃｍ,至部分神经纤维外露。紧密对合两切开面后缝合束膜、外腊。对照侧：腓总神经在相同部位切除０．５ｃｍ,至部分     

复合神经动作电位与神经再生纤维数目之间的相关性研究

目的 研究大鼠正中神经切断缝合后神经再生期间,其复合神经动作电位( CNAP)与神经再生纤维数目之间的相关性.方法 大鼠上臂正中神经中段切断后,用6种不同型号的缝线进行缝合,切断缝合后2周和8周,进行CNAP检测,观察其参数变化,然后切取大鼠正中神经组织标本(正常、缝合口近端、缝合口远端),用半薄切片甲苯胺蓝染色、免疫组化、luxol fast blue染色三种形态学检测方法进行有髓神经纤维计数,用线性回归方法分析他们之间的相关性.结果 神经切断缝合术后2周和8周,再生神经记录的CNAP波幅与正常的CNAP相比,差异有统计学意义(P＜0.01).波幅面积(Area)也显著低于正常(P＜0.01).不同型号缝线的各组之间的正中神经有髓纤维再生恢复率之间的差异都有统计学意义(P＜0.05).线性回归分析,远端记录的CNAP波幅和有髓纤维的计数之间有线性关系,拟合的线性方程,即2周:y=250.859x+ 27.826;8周:y=265.584x+2721.462.结论 大鼠正中神经切断缝合后早期和成熟期,再生神经远端的CNAP波幅与相应的有髓纤维计数之间均有线性关系,线性方程斜率的相近,仅截距有明显差异,可以在神经损伤后再生早期或晚期,根据电生理CNAP的波幅,通过线性方程计算出损伤远端的有髓纤维计数,确定神经再生的轴索通畅率,定量评定损伤神经的功能.     

Vascularized synovial flap promoting regeneration of the cryopreserved meniscal allograft: experimental study in rabbits

 The purpose of this investigation was to evaluate whether a vascularized or free synovial flap or a fibrin clot can promote regeneration of meniscal allograft in the rabbit. Seventy-eight mature New Zealand white rabbits were used. The harvested medial meniscus for the allotransplantation was frozen and stored at −80°C for 2 weeks. After resecting the medial meniscus, an allogenic meniscus was transplanted in the anatomical position (group A). The surface of the graft was covered by a vascularized synovial flap (group B), a free synovial flap (group C), or a fibrin clot (group D). The animals were killed 4, 6, 8, 12, and 16 weeks after transplantation, and the transplants were examined by gross inspection, histology, and microangiography. Connective tissue infiltration into the matrix of the graft was found to begin at 6 weeks (2/5 menisci) and to be complete at 8 weeks (5/5 menisci) in group B, whereas it began at 8 weeks (1/5) in group A. The newly formed repair tissue developed from the synovial tissue and consisted of connective tissue at the beginning and fibrocartilage later. The fibrocartilage had appeared at 8 weeks (3/5) in group B but not yet in group A (0/5). A free flap or fibrin clot coverage resulted in delayed revascularization compared to a vascularized synovial flap, but both tended to achieve faster revascularization than the controls. We concluded that regeneration of allografted meniscus with a vascularized synovial flap occurs earlier than under other conditions. Thus, allografts with synovial implantation may be considered for management of the meniscectomized knee. Received: October 18, 2001 / Accepted: September 27, 2002 Offprint requests to: K. Yamazaki     

靶肌肉注射三磷酸腺苷对周围神经再生作用的实验研究

目的通过对大鼠靶肌肉注射不同剂量的三磷酸腺苷(adenosine triphosphate，ATP)，观察其对神经再生的作用并为临床应用提供实验室依据。方法健康雌性Wistar大鼠48只，横断其左侧坐骨神经伤后立即作外膜缝合术。按手术先后随机分成3组，每组16只。高剂量组：靶肌肉注射ATP 0．5mg(0．2ml)。低剂量组：注射ATP 0．02mg(0．2ml)。对照组：注射同体积的生理盐水。术后每日用药1次至取材为止。于术后4周、8周时每组各取8只大鼠，取左侧三头肌称肌湿重。术后8周时同时检测坐骨神经的运动神经传导速度(MNCV)及形态学观察神经再生情况。结果高剂量组的所有指标均优于低剂量组，但高、低剂量组的指标均优于对照组。结论靶肌肉注射ATP对周围神经损伤有治疗作用；高剂量ATP可以发挥更好的作用。     

神经损伤(移位)时近侧分支保留对神经再生肌肉功能恢复影响的比较实验研究

目的探讨神经移位时保留神经干近侧分支对神经再生和肌肉功能恢复的影响.方法SD大鼠36只,随机分两组,实验组(分支保留组):在右侧副神经发出2个进入斜方肌肌支后3mm处,用特制的损伤钳,以相同挤压力度及相同时间将副神经夹成2mm的损伤段;对照组(分支切断组):副神经远侧段的处理与实验组相同,并将副神经近侧的两个分支切断.术后2、4、6周分别进行电生理、组织学和图像分析检测,比较近侧分支保留与否对神经再生、肌肉功能恢复的影响.结果神经干电位传导速度(NCV)恢复率、有髓神经纤维通过率、神经纤维轴突截面积恢复率的检测结果显示实验组恢复优于对照组,差异有统计学意义(P＜0.05).结论神经损伤(移位)时保留近侧分支有利于神经损伤修复(移位)术后神经再生和肌肉功能恢复.