小肠粘膜下层(SIS)桥接周围神经缺损的实验研究

目的 应用小肠粘膜下层(SIS)桥接周围神经缺损并观察结果。方法 选取健康的SD大鼠30只，随机分成三组，每组10只。先造成坐骨神经10mm缺损，然后分别用SIS桥接、自体神经移植和旷置不做处理，即SIS桥接神经组、自体神经移植对照组和空白对照组。分别于术后6周和10周取材，通过病理组织检查、神经电生理检查和计算机图像分析来评价神经再生。结果 SIS桥接神经组可见有再生神经组织长过缺损，呈条索状连续样，且神经纤维多集中在SIS形成的桥接管周缘区域，而中心区域可见胶原组织和孔隙较多。从电生理检查和计算机图像分析结果来看，SIS桥接神经组比自体神经移植组略差。结论 SIS具有促进周围神经轴突再生的作用，有望成为自体神经的替代材料应用于周围神经缺损的修复。     

自体神经外膜小间隙桥接法构建神经再生室修复周围神经断裂的实验研究

目的 探讨应用自体神经外膜小间隙桥接法构建神经再生室修复周围神经断裂的可行性和优越性.方法 取SD大鼠40只,体质量250～350g,随机分成2组,每组20只.实验组:坐骨神经切断后采用自体神经外膜小间隙桥接法缝合;对照组:坐骨神经切断后采用神经外膜原位缝合.术后2、4、6、8、10、12、14及16周时,分别从2组中各取出2只大鼠,大体观察实验动物肢体恢复自主活动的时间,显微镜视下观察神经再生室及缝合口的大体形态,光镜及电镜下对2组再生神经纤维进行观察.结果 实验组大鼠恢复肢体自主活动快,缝合口与周围组织粘连轻.形态学观察发现小间隙神经纤维排列有序,结缔组织增生明显减少.电镜观察显示神经间隙远端再生神经髓鞘板层呈同心圆状排列致密、整齐,雪旺细胞增生明显.而对照组没有上述改变.结论 自体神经外膜小间隙桥接法构建神经再生室修复周围神经断裂是可行的,优于神经外膜原位缝合法,是修复周围神经断裂的一种新方法.     

构建生物人工神经修复周围神经缺损的实验研究

目的：探讨生物人工神经对周围神经再生的作用。方法：用自体雪旺氏细胞、Ⅳ型胶原及可降解的聚乳酸导管构建生物人工神经，桥接10mm缺损的鼠坐骨神经，对照组行自体神经移植，术后8周，2组行大体观察、组织学检查及神经电生理检查。结果：实验组再生神经纤维数、再生轴突恢复率、运动神经传导速度及复合肌肉动作电位振幅均同对照组相近。结论：实验构建的生物人工神经能有效的促进周围神经再生。     

硅胶管神经间架桥的神经再生观察

硅胶管神经间架桥的神经再生观察刘建华廉冬梅用各种材料桥接周围神经缺损的方法很多。我们对一例硅胶管神经间架桥患者施行再次手术时发现，长段神经缺损用硅胶管桥接，神经间无再生现象。报道如下。１病例介绍患者男，３１岁。１９９４年１月６日因右腕部被拖拉机皮...     

小间隙影响周围神经再生超微结构特征的对比研究

目的 :利用周围神经营养、趋化性套接周围神经缺损 ,找出一个使周围神经自行修复的最佳间隙 ,比较其再生神经超微结构特征。方法 :将 40只大鼠的双侧坐骨神经外露于 10倍显微镜下 ,实验侧部分切除坐骨神经后留有 3 .0、5 .0、7.0和 10 .0mm的间隙用预制的动脉分别套接坐骨神经两端 ,对照侧坐骨神经切断后直接吻合 ,8周后各组行透射电镜观察。结果 :3 .0、5 .0mm小间隙神经再生最好 ,7.0mm再生差 ,而 10 .0mm再生神经没达到远端     

神经生长因子与睫状神经营养因子促进大鼠坐骨神经再生的协同作用研究

目的:利用新型神经再生小室探讨神经生长因子(nerve growth factor,NGF)和睫状神经营养因子(ciliary neurotrophic factor,CNTF)促进周围神经再生的协同性作用。方法:36只大鼠随机分为A、B、C3组,每组12只,用自行研制的梭形双通道桥接管桥接坐骨神经10mm缺损。A组,2支管内均注入几丁糖凝胶(100μl);B组,一侧支管内注入几丁糖(100μl) NGF(5μl) 生理盐水(5μl),另一侧支管内注入几丁糖(100μl) NGF(5μl) CNTF(5μl);C组,一侧支管内注入几丁糖(100μl) CNTF(5μl) 生理盐水(5μl),另一侧支管内注入几丁糖(100μl) NGF(5μl) CNTF(5μl)。术后8、16周对再生神经的大体形态、常规病理及超微结构进行观察,并用核黄逆行示踪评估再生神经的轴浆运输功能。结果:NGF CNTF侧再生神经呈淡黄色,质韧,弹性佳,优于对照侧,形态学观察见再生神经纤维数目多,粗细均匀,排列整齐,有髓纤维髓鞘厚,轴突直径大。其轴浆运输功能也明显优于对照侧。结论:NGF与CNTF对促进周围神经形态结构和功能的恢复均具有明显的协同作用。     

FK506缓释剂促进周围神经再生的形态学研究

[目的]探讨FK506缓释剂对周围神经再生纤维形态学恢复的促进作用.[方法]利用自行研制的梭形双通道桥接管桥接32只SD (Sprague-Dawley)大鼠坐骨神经长10 mm缺损.根据梭形管的两支管内加入的药物不同将动物随机均分为A组(两支管内均加入几丁糖凝胶)、B组(一侧支管内注入几丁糖+ FK506(B1组);另一侧支管内注入几丁糖(B2组)).术后8、16周对再生神经进行Holmes银染、透射电镜观察、硫代乙酰化胆碱染色和碳酸酐酶染色观察再生神经形态结构.[结果]术后8、16周,Holmes还原银染色及透射电镜观察到A组两支管内再生纤维无明显差异,与B2组比,B1组再生纤维数量多,排列整齐,粗细均匀;其运动纤维与感觉纤维的数量均优于对照组(P＜0.05).[结论] FK506缓释剂可明显促进周围神经再生纤维形态结构的恢复.     

胶原蛋白-明胶支架材料修复大鼠坐骨神经缺损的研究

目的制备胶原蛋白一明胶神经支架材料（CG材料）并研究其在修复大鼠10mm坐骨神经缺损实验中的疗效。方法以I型胶原蛋白、明胶通过冷冻干燥技术制备具有轴向微管结构的神经支架材料，用其桥接修复sD大鼠坐骨神经10mm缺损。术后16周分别行透射电镜，S-100、β-tubulin class Ⅲ、NFl60免疫荧光染色以及电生理检测，观察支架材料引导神经再生的疗效。结果制备的材料内部为孔径均匀且平行排列的微管结构，术后16周通过透射电镜和免疫荧光染色可见大量再生神经纤维，电生理检测神经传导速度及波幅接近自体神经移植。结论胶原蛋白一明胶支架材料能够有效的促进周围神经再生。     

应用激活态的雪旺细胞填充导管修复周围神经缺损的初步研究

目的　了解激活态的雪旺细胞在修复周围神经缺损中的作用。方法　将具有激活态的雪旺细胞填充硅管后修复大鼠前臂正中神经缺损 (12mm)。并用新鲜神经移植或单纯硅管修复作对照。术后 12周检测大鼠前臂屈肌的肌肉湿重和肌张力 ,用增殖细胞核抗体 (proliferationcellnuclearantigen ,PCNA)免疫组化方法标记。同时用计算机图像分析和S 10 0蛋白及单克隆抗体神经丝荧光免疫组化方法标记再生的神经。结果　激活态组的神经再生和肌张力的恢复明显好于对照组 ,两者差异均有显著性意义(P <0 .0 5 ,P <0 .0 1)。激活态组肌肉湿重的恢复率为 (88.2 3± 6 .0 8) % ;肌肉最大收缩张力的恢复率为 (10 1.34± 47.6 2 ) % ;肌收缩张力的恢复率为 (93 .48± 15 .72 ) % ;PCNA表达再生肌细胞最多。计算机图像分析激活态组神经再生轴突数为 (3 841± 5 2 2 )个 /条 ;再生轴突面积平均值为 2 3 .70 μm2 。S 10 0蛋白及单克隆抗体神经丝荧光免疫组化方法标记 ,证实激活态组再生的神经生长最好。结论　应用激活态的雪旺细胞填充硅管来修复正中神经缺损 ,术后神经再生和肌张力的恢复比自体神经移植和单纯硅管修复效果要好 ,为修复周围神经缺损提供了一个新的好方法     

静脉内填入神经段桥接周围神经缺损的研究

非神经移植材料修复周围神经缺损的实验研究与临床应用近年来取得一定的效果。但尚存在很多问题，最主要的是其长度问题，我们采用中国本兔２１只，随机分三组，每组７只１４条神经，实验组采用静脉内填入两段０．３ｃｍ神经段桥接腓总神经４ｃｍ缺损。对照组用静脉直接桥接腓总神经４ｃｍ缺损。自体神经移植组将腓总神经切取４ｃｍ倒置后回植，术后经２５周的观察发现实验组与自体神经移植组二者在肢体运动恢复，肌肉动作电位，神经传导速度及组织切片上的神经纤维及轴索的再生均极为相似。而用静脉直接桥接腓总神经缺损对照组失败。本实验初步表明长段静脉内填入神经段桥接周围神经缺损的可行性。     

Local effect of celecoxib on peripheral nerve repair com- bined with silicone tubulization in rat

Objective： To assess local effect of celecoxib on nerve regeneration in a rat sciatic nerve transec- tion model. Methods： Forty-five male healthy white Wistar rats were randomly divided into three experimental groups （n= 15 for each）： sham-operation （SHAM）, control （SIL） and celecoxib treated （SIL/CLX） groups. In SHAM group after anesthesia left sciatic nerve was exposed and after homeo- stasis muscle was sutured. In SIL group the left sciatic nerve was exposed in the same way and transected proximal to tibioperoneal bifurcation leaving a 10 mm gap. Proximal and distal stumps were each inserted into a silicone tube and filled with 10 gl phosphate buffered solution. In SIL/CLX group defect was bridged using a silicone tube filled with 10 μl celecoxib （0.1 g/L）. Results： Functional study and gastrocnemius muscle mass confirmed faster and better recovery of regenerated axons in SIL/CLX than in SIL group （P〈0.05）. Morphometric indices of regenerated fibers showed number and diameter of the myelinated fibers in SIL/CLX were significantly greater than those in control group. In immunohistochemistry, lo- cation of reactions to S-100 in SIL/CLX was clearly more positive than that in SIL group. Conclusion： Response to local treatment ofcelecoxib demonstrates that it influences and improves functional re- covery of peripheral nerve regeneration.     

大鼠坐骨神经损伤修复后近端轴突再生的初步研究

目的 采用生长相关蛋白-43(GAP-43)对SD大鼠坐骨神经近端新生轴突进行标记,观察坐骨神经损伤修复后近端轴突再生过程.方法 64只SPF级SD大鼠于右侧坐骨神经分叉以上5mm处切断坐骨神经,分别采用原位缝合以及生物可溶性甲壳质套管小间隙(2 mm)缝合方法进行修复,分别于术后1、3、7及14 d取材.观察神经缝合处组织大体形态;套管内坐骨神经生长状态;近端新生轴突形态,并应用图像分析方法对新生轴突数目进行定量分析.结果 套管小间隙缝合组大鼠神经缝合处粘连情况较轻.免疫荧光染色显示:术后14 d,新生纤维神经干形成圆锥形向前生长.形态规则均一.原位缝合组大鼠缝合处粘连较重,新生轴突于7 d左右长过缝合点,坐骨神经远端新生轴突散在分布,形态不规则.免疫荧光染色图像分析结果显示:大鼠坐骨神经损伤修复后,术后3 d原位缝合组新生轴突数目较套管缝合组多.差异有统计学意义(P<0.01);术后7 d套管小间隙缝合组大鼠坐骨神经开始大量再生;术后14 d左右,套管小间隙缝合组新生轴突数目显著高于原位缝合组(P<0.05).结论 甲壳质套管具有良好的生物相容性,套管内新生轴突前沿以圆锥形向前生长,新生轴突形态较原位缝合组好,套管小间隙修复7 d后新生轴突数目开始较原位缝合组多.本实验主要观察了两种术式修复后大鼠坐骨神经新生轴突的生长规律,也从组织学角度解释了生物套管小间隙套接方法的再生效果比原位缝合效果好的可能原因所在.     

人发角蛋白桥接周围神经缺损后的相容性和降解性的实验研究

目的探讨人发角蛋白(H H K)在桥接周围神经缺损后,诱导神经再生及与周围组织的相容和自身的降解情况。方法用聚乳酸-羟基乙酸(P LG A)套管装配H H K来桥接SD大鼠10m m长的坐骨神经缺损,术后第3、6、9、12周取材,切取中段行H E染色,并对第6周标本加行S100蛋白和神经丝(N F)免疫组化染色,12周标本横切加行透射电镜检查,做组织学观察。结果术后3周H H K毛小皮已经开始剥脱降解,并已有大量的雪旺细胞和神经纤维沿H H K长入,6周时H H K均已质化,免疫组化染色见大量的S100蛋白阳性细胞和N F阳性纤维;9、12周H H K继续降解,长入其间的雪旺细胞和神经纤维更多,排列更加规律,电镜下见已形成较厚的髓鞘板层。结论H H K具有很好的生物相容性和可控的降解速率,能很好的诱导神经再生,可作为支架材料应用于周围神经组织工程研究。     

甲壳质套管桥接周围神经长度极限的研究

目的 通过采用不同间隙套接修复大鼠坐骨神经损伤的实验研究,探讨甲壳质套管桥接修复周围神经损伤的长度极限.方法 SPF级健康成年雄性SD大鼠24只,于大鼠右侧坐骨神经分叉处以上5 mm建立坐骨神经离断伤模型,部分切除坐骨神经后使用甲壳质套管桥接修复,使神经断端间留有2、5、8和10 mm间隙.8周后常规锇酸染色,镜下观察套管远端有髓神经纤维数目、轴突平均面积、髓鞘平均厚度及腓肠肌平均湿重,并进行定量组织学分析.结果 术后8周显示2 mm小间隙组神经纤维已有80%再生,再生效果最佳,与其他组相比差异有统计学意义(P＜0.01).随着间隙距离逐渐增加,神经纤维再生效果逐渐变差;5 mm间隙组再生约60%,效果优于8 mm间隙组(P＜0.01);8 mm间隙组再生约20%,优于10 mm间隙组(P＜0.01);10 mm间隙组只有1%有髓神经纤维成功长入远端,肌肉湿重降至正常的42.9%.结论 使用甲壳质套管桥接修复大鼠坐骨神经损伤时,2 mm左右小间隙套接修复效果最佳,5 mm间隙是修复后周围神经功能得到恢复的极限间隙,10 mm是神经单靠趋化性、营养作用再生的最大间隙.     

Acetyl Salicylic Acid Locally Enhances Functional Recovery after Sciatic Nerve Transection in Rat

Local effect of acetyl salicylic acid (ASA) on peripheral nerve regeneration was studied using a rat sciatic nerve transection model. Forty-five male healthy White Wistar rats were divided into three experimental groups (n = 15), randomly: Sham-operation (SHAM), control (SIL), and ASA-treated (SIL/ASA) groups. In SHAM group after anesthesia left sciatic nerve was exposed through a gluteal muscle incision and after homeostasis the muscle was sutured. In SIL group the left sciatic nerve was exposed the same way and transected proximal to tibio-peroneal bifurcation leaving a 10-mm gap. Proximal and distal stumps were each inserted into a silicone tube and filled with 10 μl phosphate buffered solution. In SIL/ASA group defect was bridged using a silicone tube filled with 10 μl acetyl salisylic acid (0.1 mg/ml). Each group was subdivided into three subgroups of five animals each and were studied 4, 8, and 12 weeks after surgery. Data were analyzed statistically by factorial analysis of variance (ANOVA) and the Bonferroni test for pair-wise comparisons. Functional study confirmed faster and better recovery of regenerated axons in SIL/ASA than in SIL group (p < 0.05). Gastrocnemius muscle mass in SIL/ASA was significantly more than in SIL group. Morphometric indices of regenerated fibers showed that the number and diameter of the myelinated fibers in SIL/ASA were significantly higher than in control group. In immuohistochemistry, location of reactions to S-100 in SIL/ASA was clearly more positive than in SIL group. Response to local treatment of ASA demonstrates that it influences and improves functional recovery of peripheral nerve regeneration.     

Nerve growth factor combined with an epineural conduit for bridging a short nerve gap (10 mm). A study in rabbits

The purpose of this study was to evaluate the effect of direct administration of nerve growth factor (NGF) into an epineural conduit across a short nerve gap (10 mm) in a rabbit sciatic nerve model. The animals were divided into two groups. In group 1, n = 6, a 10-mm defect was created in the sciatic nerve and bridged with an epineural flap. A dose of 1 μg of NGF was locally administered daily for the first 21 days. NGF administration was made inside the epineural flap using a silicone reservoir connected to a silicone tube. In group 2, n = 6, the 10-mm defect was bridged with a nerve graft. This group did not receive any further treatment. At 13 weeks, all animals, before euthanasia, underwent electromyography (EMG) studies and then specimen sent for histology morphometric analysis. NGF administration ensured a significantly increased average number of myelinated axons per μm(2) (P = 0.028) and promoted fiber maturation (P = 0.031) and better EMG results (P = 0.046 for latency P = 0.048 for amplitude), compared with the control group. Although nerve grafts remain the gold standard for peripheral nerve repair, NGF-treated epineural conduits represent a good alternative, particularly when an unfavorable environment for nerve grafts is present.     

Promoting nerve regeneration through long gaps using a small nerve tissue graft

BACKGROUND: If nerve tissue is capable of inducing regeneration, as suggested by the neurotropism theory, then even small pieces of nerve tissue should have the potential to induce nerve regeneration. Therefore, long gaps might presumably be bridged via the neurotrophic potential of small pieces of nerve tissue grafted into the middle of the nerve gap. It is necessary to confirm the validity of the neurotropism theory and to also explore the potential usefulness of small nerve grafting through long gaps. METHODS: A small piece of nerve tissue was grafted into a silicone tube bridging a relatively long nerve gap in an attempt to promote nerve regeneration. A 15-mm gap was created in the left sciatic nerve of 31 Wistar rats (8 weeks of age). The experimental groups included one with nonvascularized nerve tissue grafted into a silicone tube with no distal nerve suturing (NV-A), another with vascularized nerve tissue grafted into a silicone tube with no distal nerve suturing (V-A), a third group with nonvascularized nerve tissue grafted into a silicone tube with distal nerve suturing (NV-P), a fourth group with vascularized nerve tissue grafted into a silicone tube with distal nerve suturing (V-P), and a group with no nerve segment grafted into the silicone tube (control). Electrophysiologic and histologic examinations were performed 10 weeks after the operation. RESULTS: No regeneration was obtained in the control group. Nerve regeneration was evident at the proximal end of the tube in the NV-A, V-A, NV-P, and V-P groups, and at the distal end in the NV-P and V-P groups. The degree of distal regeneration was extremely slight in the NV-A and V-A groups. An electrophysiologic examination performed in the NV-P and V-P groups revealed better results in the latter group. CONCLUSION: Small nerve grafts are capable of inducing nerve regeneration even over a long nerve gap, by grafting nerve tissue into the middle of the lesion using a silicone tube.     

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。     

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.     

Stromal vascular fraction combined with silicone rubber chamber improves sciatic nerve regeneration in diabetes

Purpose: To study the effects of transplantation of characterized uncultured stromal vascular fraction (SVF) on sciatic nerve regeneration. Methods: A 10-mm sciatic nerve defect was bridged using a silicone conduit filled with SVF. In control group, silicone conduit was filled with phosphate-buffered saline alone. In sham-operated group, the sciatic nerve was only exposed and manipulated. The regenerated nerve fibers were studied 8 and 12 weeks after surgery. Results: Behavioral and functional studies confirmed faster recovery of regenerated axons in SVF transplanted animals than in control group (p < 0.05). Gastrocnemius muscle mass in SVF transplanted animal was found to be significantly more than that in control group. Morphometric indices of the regenerated fibers showed the number and diameter of the myelinated fibers to be significantly higher in SVF transplanted animals than in control group. In immunohistochemistry, the location of reactions to S-100 in SVF transplanted animals was clearly more positive than that in control group. Conclusion: SVF transplantation combined with silicone conduit could be considered as a readily accessible source of stromal cells that improves functional recovery of sciatic nerve. It may have clinical implications for the surgical management of acute diabetic patients after facial nerve transection.