Hsp27联合ADSC对脱细胞神经支架修复小鼠坐骨神经缺损后caspase-3的影响

目的探讨热休克蛋白27(Hsp27)与脂肪源性干细胞(ADSC)联合应用对小鼠脱细胞同种异体神经支架移植后凋亡相关基因caspase-3表达及功能恢复的影响。方法 30只成年C57BL/6小鼠随机分为脱细胞神经支架(ANA)组、ADSC组和Hsp27+ADSC组。坐骨神经功能指数(SFI)、电生理和胫前肌湿重比方法检测神经运动功能的恢复,Western bolt检测神经支架内Hsp27和caspase-3的蛋白表达。胆碱乙酰转移酶(Ch AT)免疫荧光法检测神经移植体中运动轴突表达,并示踪PKH26标记的移植ADSC。结果与ADSC组比较,Hsp27+ADSC组神经移植体内Hsp27蛋白表达显著增高,caspase-3蛋白表达明显降低,Ch AT标记运动轴突表达增强,PKH-26标记的ADSC数量显著增高(P0.05)。与ANA组比较,ADSC组和Hsp27+ADSC组SFI增高、电生理波幅增高、神经传导速度增快、延迟期缩短、胫前肌湿重比率增高,其中Hsp27+ADSC组神经恢复作用显著优于ADSC组(P0.05)。结论Hsp27联合ADSC移植对小鼠脱细胞异体神经支架修复坐骨神经缺损后运动轴突再生和功能恢复的作用优于ADSC组,其机制可能与Hsp2抑制caspase-3凋亡信号通路,促进移植的ADSC存活有关。     

小鼠神经缺损管式修复后长时程功能恢复与再生评价

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。     

富血小板血浆诱导骨髓间充质干细胞结合化学萃取去细胞神经修复坐骨神经缺损

背景:周围神经损伤早期许旺细胞尚未大量分裂增殖,此时由于解剖连续性的中断,通过轴浆逆向运输提供的营养因子骤减,缺乏神经营养因子支持的神经元有可能死亡,从而使周围神经不能再生或再生乏力。目的:观察植入经富血小板血浆诱导的骨髓间充质干细胞结合去细胞神经修复坐骨神经缺损的效果。方法:取新西兰大耳白兔制备坐骨神经缺损模型,随机抽签法分成4组:去细胞神经组,移植同种异体去细胞神经;骨髓间充质干细胞组,移植同种异体骨髓间充质干细胞结合化学萃取的同种异体去细胞神经:经诱导骨髓间充质干细胞组,移植经富血小板血浆诱导的同种异体骨髓间充质干细胞结合化学萃取的同种异体去细胞神经;自体神经组,移植自体神经。术后进行形态学观察与靶肌肉肌湿质量恢复率、运动神经传导速度、轴突直径和髓鞘厚度的检测。结果与结论:经富血小板血浆诱导的骨髓间充质干细胞结合化学萃取的去细胞神经移植修复神经的靶肌肉肌湿质量恢复率、运动神经传导速度、轴突直径和髓鞘厚度及形态学观察明显优于移植单纯化学萃取的去细胞神经与骨髓间充质干细胞结合化学萃取的去细胞神经的效果,而与移植自体神经修复结果相似。说明经诱导后的骨髓间充质干细胞在体内具有许旺细胞的部分功能,可作为组织工程化外周神经的种子细胞,用于周围神经缺损的修复。     

脱细胞同种异体神经移植物修复大鼠坐骨神经缺损的实验研究

目的 观察脱细胞处理的同种异体神经移植物修复大鼠坐骨神经缺损的作用。方法 用组织工程学方法制备的大鼠同种异体神经移植物桥接大鼠坐骨神经缺损，并对再生神经进行电生理学功能测试，光镜、电镜观察移植物内的再生神经，并进行统计分析。结果 术后13周内，动物未见炎症及排斥反应。用脱细胞处理的同种异体神经移植物修复神经缺损，再生神经的传导功能、纤维数量、轴突直径、有髓纤维占有的面积与自体神经移植对照及计量学上统计分析均无显著性差异。结论 脱细胞处理的同种异体神经移植物具有良好的组织相容性，它对缺损的坐骨神经再生有促进作用。     

脱细胞同种异体神经移植物促进大鼠运动功能恢复的实验研究

目的　检测脱细胞同种异体神经移植物桥接大鼠坐骨神经 10mm缺损后运动功能的恢复。方法　用脱细胞同种异体神经移植物桥接大鼠坐骨神经 10mm缺损 ,术后 12周、16周运用电生理及AchE结合镀银染色检测再生神经传导速度和腓肠肌的运动终板。自体神经移植作为对照组。结果　实验组术后 12周、16周再生神经传导速度与对照组相比 ,无显著性差异 ;术后 12周组织化学染色可见腓肠肌内有呈AchE阳性的运动终板 ;16周运动终板AchE阳性反应加深 ,并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。结论　脱细胞同种异体神经移植物桥接大鼠坐骨神经缺损具有促进其运动功能恢复的作用     

坐骨神经预损伤对脊髓损伤后小鼠神经功能恢复的影响

目的研究坐骨神经预损伤对脊髓损伤后小鼠神经功能恢复的影响。方法以C57BL/6小鼠为研究对象,分为假手术组(对照组,Control),脊髓损伤(spinal cord injury, SCI)组和坐骨神经预损伤(sciatic nerve injury,SNI)组。假手术组仅移除小鼠T8-T10椎骨上棘暴露胸髓,SCI组移除小鼠T8-T10椎骨上棘暴露胸髓给予外力打击造成标准损伤,SNI组小鼠手术切断右腿坐骨神经,在预损伤7d后进行脊髓损伤。在恢复期(即脊髓损伤14d后),通过电生理实验检测运动神经传导功能,通过组织病理学检验检测组织缺损恢复、髓鞘再生及小胶质细胞的募集活化。结果电生理实验表明SNI组小鼠运动传导功能恢复明显强于SCI组;HE染色结果说明SNI组脊髓组织缺损恢复情况明显强于SNI组;以MBP为指标的免疫荧光结果显示SNI组髓鞘含量明显多于SCI组,髓鞘再生能力明显增强;以IBA-1为指标的免疫荧光结果显示SNI组小胶质细胞明显增多,坐骨神经预损伤明显增强了小胶质细胞的募集活化作用。结论坐骨神经预损伤处理可促进小鼠脊髓损伤后组织形态恢复、小胶质募集活化作用、髓鞘再生作用和神经传导功能的恢复。     

NGF和GM_1联合应用对去细胞异种神经支架修复大鼠坐骨神经陈旧性缺损的影响

目的:研究神经生长因子(NGF)和单唾液酸四己糖神经节苷脂(GM1)联合应用对去细胞异种神经支架修复大鼠坐骨神经陈旧性缺损后神经再生和功能恢复的作用。方法:制作大鼠坐骨神经陈旧性缺损模型,用去细胞异种神经支架进行桥接修复。大鼠术侧小腿三头肌内分别注射NGF+GM1液或NS液,每日1次连续2周。于修复术后8或14周,检测计算大鼠坐骨神经运动传导速度(MNCV)恢复率和小腿腓肠肌复合动作电位(CMAP)波幅恢复率;辣根过氧化物酶(HRP)作为逆行神经轴突示踪剂注射于损伤神经,观察脊神经节内神经元HRP标记情况;检测小腿三头肌湿重及腓肠肌纤维平均截面积恢复率;免疫荧光染色等方法观察移植的神经支架内及其近、远端吻合口的神经纤维组织学特点。结果:在修复术后8周和12周的动物中都观察到:NGF+GM1组动物的MNCV恢复率、CMAP波幅恢复率、L3~L5脊神经节HRP标记细胞数、小腿三头肌湿重恢复率及腓肠肌纤维平均截面积恢复率均优于NS对照组(P﹤0.05);NGF+GM1组动物移植神经支架内再生神经纤维更加密集、排列规则整齐,神经支架内Schwann细胞(SC)大量增殖。结论:结果表明NGF及GM1联合去细胞异种神经支架可成功地修复周围神经陈旧性缺损,促进神经再生和功能恢复。     

去细胞神经与自体神经移植桥接修复大鼠坐骨神经缺损的对比研究

目的观察同种异体去细胞神经与自体神经移植桥接修复大鼠坐骨神经缺损的神经再生情况。方法制备大鼠同种异体去细胞神经及大鼠坐骨神经缺损模型,修复12周后应用HE染色,Bielschowsky改良染色,Weil氏铁明矾苏木素染色,光镜下观察神经外膜上的微血管数和微血管面积百分比,计数单位面积的轴突数目,远端轴突密度/近端轴突密度为再生神经通过率,计数单位面积的有髓神经纤维数目和有髓神经纤维的直径。结果在坐骨神经纤维的再生神经纤维通过率、有髓神经纤维密度和直径、桥接体微血管的数目和微血管面积百分比等再生指标上,自体神经移植组略优于化学去细胞神经组。结论同种异体去细胞神经移植可促进神经再生,但仍然不如自体移植效果好。     

冷冻保存胎兔周围神经异体移植

目的 :探讨胚胎兔周围神经经冷冻保存后 ,同种异体移植修复周围神经缺损的作用。方法 :以采用两步冷冻方法制备冷冻保存的胎兔坐骨神经 (维持温度值 - 4 0℃ )为供体 ,移植桥接成年兔腓总神经缺损区 ,并与自体神经移植、新鲜异体神经移植、空白缺损比较 ,16周后作再生神经电生理检测、神经断面图像分析及透射电镜观察。结果 :胎神经移植组神经再生效果不如自体神经好 ,但优于新鲜异体神经组。结论 :冷冻保存胎神经移植桥接周围神经缺损 ,可以促进和引到轴突再生 ,有可能代替自体神经移植     

异体骨髓间充质干细胞移植治疗大鼠脊髓损伤

背景:脊髓损伤的修复目前尚无良好的治疗手段,细胞移植能促进神经轴突再生及脊髓功能恢复,为治疗脊髓损伤提供了可能,但因脊髓损伤模型及移植方式不同,其治疗效果并不相同。目的:验证异体骨髓间充质干细胞移植对大鼠脊髓损伤的治疗作用。方法:全骨髓贴壁法分离大鼠骨髓间充质干细胞。健康SD大鼠随机分为3组,细胞移植组、对照组和假手术组。细胞移植组和对照组采用改良Allen重物打击法制造大鼠脊髓损伤模型,假手术组仅暴露脊髓。术后4周,每周进行运动功能评分ELISA检测脊髓损伤组织中脑源性神经营养因子、神经生长因子表达;免疫荧光染色检测脊髓组织中NF200和胶质纤维酸性蛋白表达。结果与结论:与对照组比较,细胞移植组大鼠运动功能明显改善,脊髓组织中脑源性神经营养因子、神经生长因子蛋白含量明显增高(P0.05);移植组大鼠脊髓囊腔较小,NF200表达明显增加,胶质纤维酸性蛋白表达减少。提示异体骨髓间充质干细胞移植能增加损伤脊髓神经生长因子含量,抑制胶质瘢痕形成,促进神经轴突再生,改善大鼠脊髓损伤后运动功能恢复。     

大鼠异体神经移植后再生神经生物力学性质研究

用Ｗｉｓｔａｒ鼠作为实验模型,切下１ｃｍ坐骨神经,再用同系Ｗｉｓｔａｒ鼠坐骨神经异体桥接,修复坐骨神经的缺损,术后２４周对Ｗｉｓｔａｒ鼠的手术侧与正常侧用指标抗张强度与弹性模量（ε＝１０％）进行测试,辅以电镜,光镜观察。     

Intramuscular Martin-Gruber anastomosis

The incidence and morphology of the intramuscular Martin-Gruber anastomosis are presented based on the study of 118 human cadavers (55 male, 63 female). The Martin-Gruber anastomosis was found in 25 (21.2%) of the 118 cadavers. It occurred in 11 (20%) of the 55 male cadavers (4 bilateral, 7 unilateral; 5 left and 2 right) and in 14 (22.2%) of the 63 female cadavers (2 bilateral, 12 unilateral; 8 left and 4 right). Therefore, the Martin-Gruber anastomosis was found in 31 (13.1%) of the 236 upper limbs. According to a recent classification (Rodríguez-Niedenführ et al., 2000), pattern I was found in 29 cases (93.5%), corresponding to Type A in 13 (41.9%), Type B in 3 (9.7%) and Type C in 13 (41.9%), whereas pattern II was found in 2 cases (6.5%), both being a duplication of Type IC. Intramuscular Martin-Gruber anastomosis was a single anastomosis that originated in all cases from the anterior interosseous nerve (pattern IC) and then passed through a muscle bundle of the flexor digitorum profundus and behind the ulnar artery to join the ulnar nerve as a single connecting branch. It did not send branches to the flexor digitorum profundus. This intramuscular course was observed in 3 of the 13 cases of Type C anastomosis (23.1%) or 3 cases out of 31 Martin-Gruber anastomoses (10%).     

bFGF对同种异体神经移植后周围神经再生的影响

目的 :探讨bFGF对同种异体神经移植后周围神经再生的影响。方法 :将反复冻融的大鼠神经移植于另一大鼠的坐骨神经 ,实验组注射bFGF 1 0 0u/d共 1 0d ,对照组注射生理盐水 1 0d。术后大鼠存活 1 2周 ,光镜下用体视学方法测试再生神经纤维的面数密度 (NA)、面积密度 (AA)、横切面面积 (AE)、脊髓前角运动细胞和脊神经节细胞的体密度 (VV)、数密度 (NV)。结果 :两组均可见再生神经纤维长入异体移植神经并向远段延伸。实验组再生神经纤维的NA、AA、脊髓前角运动细胞和脊神经节细胞的VV、NV 与对照组的比较 ,有显著性差异。结论 :bFGF能促进周围神经再生 ,对脊髓前角运动细胞和脊神经节细胞的存活有保护作用。     

Martin-Gruber anastomosis revisited

Based on a study of 70 human cadavers (31 male, 39 female) and on cases described previously, we propose a new classification of the Martin-Gruber anastomosis, a neural connection between the median and ulnar nerves in the forearm. The anastomosis was found in 16 (22.9%) cadavers, being bilateral in three (18.7%) and unilateral in 13 (81.3%), five right and eight left. It occurred in eight (25.8%) of the 31 male cadavers and in eight (20.5%) of the 39 females. Therefore, the anastomosis was found in 19 (13.6%) of the 140 forearms. In Pattern I (89.5%) the anastomosis was made by only one branch, whereas in Pattern II (10.5%) it was made by two. The individual branches were classified as Types a, b, and c based on the nature of their origin from the median nerve. Type a (47.3%) arose from the branch to the superficial forearm flexor muscles, Type b (10.6%) from the common trunk, and Type c (31.6%) from the anterior interosseous nerve. Pattern II was a duplication of Type c (10.5%). The anastomotic branch took an oblique or arched course before joining the ulnar nerve, undivided in 15 cases, but divided into two branches in four cases. The anastomosis passed in front of the ulnar artery in four cases, behind it in six, and in nine cases it was related to the anterior ulnar recurrent artery.     

Sural-tibial nerve communications in humans

We investigated the occurrence of a communication between the sural and tibial nerves in 49 legs of 28 Japanese cadavers. In front of the calcanean tendon, we found the communication in 7 legs (14.3%) or in 5 cadavers (18.9%). The sural nerve gave rise to a number of medial and lateral branches, including the lateral calcanean branch at the lateral side of the ankle. The communicating branch with the tibial nerve arose from the first medial branch and pierced the deep fascia of the leg. In 4 cases, the U-shaped communication was formed between the sural and tibial nerves, and in 3 cases, the Y-shaped communication. Electrophysiologi-cal evidence of an anomalous motor function of the sural nerve has been reported recently. We consider that the U-shaped communication between the sural and tibial nerves gives a morphological basis to the motor function of the sural nerve.     

The normal and variant clinical anatomy of the sensory supply of the orbit

Orbital and retro‐orbital pain are relatively common clinical conditions that are associated with such disorders as trigeminal, lacrimal, and ciliary neuralgia, cluster headaches, paroxysmal hemicrania, inflammatory orbital pseudotumor, trochleitis, and herpetic neuralgia ophthalmicus, thus making the nerves supplying the orbit of great clinical importance. Surprisingly, how pain from this region reaches conscious levels is enigmatic. Classically, it has been assumed that pain reaches the ophthalmic division of the trigeminal nerve (V₁) and travels to the descending spinal trigeminal nucleus. However, exactly where the receptors for orbital pain are located and how impulses reach V₁ is speculative. In this project, we reviewed all of the reported connections between the orbital nerves and V₁ in order to understand how pain from this region is transmitted to the brain. We found reported neural connections to exist between cranial nerve (CN) V₁ and CNs III, IV, and VI within the orbit, as well as direct neural branches to extra‐ocular muscles from the nasociliary, frontal, and supraorbital nerves. We also found reported neural connections to exist between the presumed carotid plexus and CN VI and CN V₁, CN VI and CN V₁ and V₂, and between CN V₁ and CN III, all within the cavernous sinus. Whether or not these connections are sympathetic or sensory or some combination of both connections remains unclear. An understanding of the variability and frequency of these neural connections could lead to safer surgical procedures of the orbit and effective treatments for patients with orbital pain. Clin. Anat. 27:169–175, 2014. © 2014 Wiley Periodicals, Inc.     

神经生长因子对周围神经损伤后再生和修复的实验研究

手术切除５ｍｍ兔的尺神经，在两断端间连接肌桥并套装硅胶管，形成一个封闭腔，向腔内注入神经生长因子。间隔不同时间取尺神经桥接区、桥接区近段、远段、尺神经的脊髓投射节段和相应脊神经节，用光镜和电镜观察神经溃变和再生情况并作图像分析；用酶标示踪和电生理方法检测神经通路的重建状况。结果显示，周围神经离断后，肌束桥接并用硅胶管套装后注入外源性神经生长因子，可明显地促进离断神经的再生和修复。     

骨间前神经转位重建鱼际肌功能

前臂或腕部正中神经断裂,直接吻合后鱼际肌功能的恢复常常令人失望,为了解决这一难题,本文在120侧成人上肢解剖学研究的基础上,采用骨间前神经转位术修复鱼际肌支9侧,获得成功.     

耳大神经移植治疗面神经麻痹的应用解剖学研究

用22具成年尸体的44侧耳大神经进行了解剖观察和测量.耳大神经的平均长度为4.12cm,有2～4个分支.耳大神经的体表投影相当于胸锁乳突肌后缘中点稍下方至耳垂的连线,主干在颈外静脉后方约0.7cm.结果表明:耳大神经是用于神经移植治疗面神经麻痹较理想的神经移植物,耳大神经的体表投影、颈外静脉及下颌角是术中寻找的可靠标志.