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。     

以化学去细胞法处理同种异体神经与周围静脉伴行修复面神经缺损

背景：有研究表明化学去细胞法处理的同种异体神经修复面神经缺损可以取得较好的修复效果。 目的：在化学去细胞法处理同种异体神经的基础上探索一种修复面神经缺损更有效的修复方式。  方法：将新西兰大白兔随机分为2组,实验组制备面神经颊支缺损动物模型,采用经化学去细胞的同种异体腓肠神经进行移植,且与伴行静脉行外膜缝合;对照组在同样位置的远近端分别切断面神经,但不破坏被切断神经与周围组织的正常解剖关系,再切断处行外膜缝合桥接。 结果与结论：修复后3个月两组兔均存活,面部表情基本对称,胡须活动正常,神经移植处未见明显瘢痕及神经瘤形成。电镜观察结果显示,实验组与对照组右侧面神经颊支传导速度,移植体远端吻合口附近5.0 mm段有髓神经纤维数量,靶肌肉运动终板计数均差异无显著性意义(P > 0.05)。结果证实,化学去细胞同种异体神经与周围静脉伴行修复家兔面神经缺损的方法可以达到与自体面神经原位移植相似的修复效果。     

冻融联合优化化学法制备粗大去细胞同种异体神经

背景：异体神经移植修复神经缺损需要面对和解决的是宿主免疫排斥反应问题。因此,如何避免、减轻免疫排斥反应是同种异体神经移植获得成功的关键因素。 目的：探索新的异体神经预处理方法,清除犬周围神经中的许旺细胞和髓鞘,保留完整的基底膜,建立粗大异体神经移植物的预处理方法,获得去细胞异体神经移植物。 方法：取健康成年杂种犬游离双侧坐骨神经,以冻融联合优化化学法对神经进行预处理,光、电镜观察其结构特征,组织学染色及Western blot分析其成分。 结果与结论：预处理后的去细胞神经的延展性和神经外膜的弹韧性良好,许旺细胞和髓鞘被彻底清除,基底膜保留完整,去细胞神经为一没有细胞、髓鞘及其碎片的空的神经基膜管。结果表明该方法有效的清除了周围神经中主要抗原成分许旺细胞及髓鞘,并且保留了促神经再生的重要成分基底膜,可作为制备组织工程化神经较理想的方法。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

单环刺缢及同种异体神经制成人工神经修复神经缺损

目的:研究去细胞的单环刺缢体壁和同种异体神经所构成的人工组织神经的组织相容性及其修复神经缺损的效果。方法:取大鼠40只随机分成实验组、自体神经组、硅胶管组、正常组。实验组将去细胞的单环刺缢体壁缝合成神经导管,其内充填去细胞的异体神经,修复大鼠10 mm坐骨神经缺损。术后4月,通过大体观察,组织形态学观察,了解该人工组织神经修复大鼠坐骨神经缺损的疗效。结果:该人工组织神经组织相容性良好,神经功能恢复效果正常组>自体神经组>实验组>硅胶管组,实验组疗效与自体神经组接近,明显优于硅胶管组。结论:将去细胞的单环刺缢体壁和同种异体神经制成人工组织神经修复周围神经缺损是可行的。     

化学去细胞同种异体神经移植修复大鼠骶1神经缺损

背景：自体神经移植是修复周围神经损伤中最常用的方法。 目的：探讨化学去细胞神经修复大鼠骶1神经缺损的效果,以期寻找修复周围神经缺损较为理想的方法和材料。 方法：取SD大鼠骶1神经,采用化学去细胞法处理大鼠骶1神经的免疫原性成分,使其成为去细胞神经。将Wistar大鼠右侧骶1神经制成1 cm缺损模型,用SD大鼠去细胞同种异体神经移植修复大鼠骶1神经的缺损。 结果与结论：与术前相比,术后8周逼尿肌漏尿点降低,膀胱最大容积和膀胱顺应性升高(P < 0.05)。神经纤维细丝蛋白染色结果显示,右侧经同种异体神经移植修复的神经吻合口断端被染成绿色,神经纤维排列整齐分布均一,再生神经轴突长入远端神经。左侧正常神经形态均一,神经纤维排列整齐分布均一未见轴突长入远端神经。结果表明,化学去细胞同种异体神经的抗原性明显降低,可修复高等哺乳类动物的周围神经损伤。     

辣根过氧化物酶逆行示踪评价组织工程人工神经修复大鼠坐骨神经缺损

背景:作者前期制备了无细胞神经移植物,并将骨髓间充质干细胞种植其内,成功构建了组织工程人工神经。目的:应用辣根过氧化物酶神经逆行示踪技术,评价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对感觉神经元的保护作用。方法:成年清洁级健康雄性SD大鼠,随机分成3组:实验组采用复合骨髓间充质干细胞的无细胞神经移植物桥接大鼠坐骨神经缺损;空白对照组采用无细胞神经移植物桥接大鼠坐骨神经缺损;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。移植后12周应用辣根过氧化物酶神经逆行示踪技术,对脊髓后根神经节感觉神经元的再生进行评价。结果与结论:移植12周后根神经节感觉神经元再生评价结果提示,实验组与自体神经对照组相比差异无显著性意义,优于空白对照组;足底皮肤S-100免疫组织化学染色结果显示,各组皮下均见棕色阳性反应。提示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,对大鼠脊髓后根神经节感觉神经元具有保护作用,可以促进感觉功能的恢复。     

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     

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。     

组织工程化脱细胞神经基质移植体的研究进展

组织工程化的细胞外基质成分、细胞或通道的基底物质已经显示出支持轴突再生和功能恢复的巨大潜能。脱细胞处理的神经基质移植体具有良好的仿生性和生物相容性,包含较丰富的促进神经生长的蛋白等成分并适合于种子细胞在其内迁移生长,移植后免疫排斥反应较低,为轴突再生提供适宜的微环境,能够有效地促进轴突再生,是修复周围神经缺损的适宜移植物,将在周围神经损伤修复中占明显优势。     

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。     

Repair of facial nerve defects with decellularized artery allografts containing autologous adipose-derived stem cells in a rat model

The purpose of this study was to investigate the effects of a decellularized artery allograft containing autologous adipose-derived stem cells (ADSCs) on an 8-mm facial nerve branch lesion in a rat model. At 8 weeks postoperatively, functional evaluation of unilateral vibrissae movements, morphological analysis of regenerated nerve segments and retrograde labeling of facial motoneurons were all analyzed. Better regenerative outcomes associated with functional improvement, great axonal growth, and improved target reinnervation were achieved in the artery-ADSCs group (2), whereas the cut nerves sutured with artery conduits alone (group 1) achieved inferior restoration. Furthermore, transected nerves repaired with nerve autografts (group 3) resulted in significant recovery of whisking, maturation of myelinated fibers and increased number of labeled facial neurons, and the latter two parameters were significantly different from those of group 2. Collectively, though our combined use of a decellularized artery allograft with autologous ADSCs achieved regenerative outcomes inferior to a nerve autograft, it certainly showed a beneficial effect on promoting nerve regeneration and thus represents an alternative approach for the reconstruction of peripheral facial nerve defects.     

Combined use of decellularized allogeneic artery conduits with autologous transdifferentiated adipose-derived stem cells for facial nerve regeneration in rats

Natural biological conduits containing seed cells have been widely used as an alternative strategy for nerve gap reconstruction to replace traditional nerve autograft techniques. The purpose of this study was to investigate the effects of a decellularized allogeneic artery conduit containing autologous transdifferentiated adipose-derived stem cells (dADSCs) on an 8-mm facial nerve branch lesion in a rat model. After 8 weeks, functional evaluation of vibrissae movements and electrophysiological assessment, retrograde labeling of facial motoneurons and morphological analysis of regenerated nerves were performed to assess nerve regeneration. The transected nerves reconstructed with dADSC-seeded artery conduits achieved satisfying regenerative outcomes associated with morphological and functional improvements which approached those achieved with Schwann cell (SC)-seeded artery conduits, and superior to those achieved with artery conduits alone or ADSC-seeded artery conduits, but inferior to those achieved with nerve autografts. Besides, numerous transplanted PKH26-labeled dADSCs maintained their acquired SC-phenotype and myelin sheath-forming capacity inside decellularized artery conduits and were involved in the process of axonal regeneration and remyelination. Collectively, our combined use of decellularized allogeneic artery conduits with autologous dADSCs certainly showed beneficial effects on nerve regeneration and functional restoration, and thus represents an alternative approach for the reconstruction of peripheral facial nerve defects.     

Tubulation with dental pulp cells promotes facial nerve regeneration in rats

Dental pulp is an easily obtainable source of viable cells for potential use in peripheral nerve regeneration. We prepared artificial conditions for nerve regeneration using a silicone tube containing a collagen gel embedded with rat dental pulp cells, and we examined its effectiveness for repairing a gap in the rat facial nerve. Twelve days after transplantation, defective facial nerves connected with silicone tubes containing dental pulp cells were repaired more rapidly than control tubes containing the collagen gel alone. When a tube containing green fluorescent protein (GFP)-positive dental pulp cells was transplanted into a facial nerve gap in a GFP-negative rat, we observed regenerated nerves with GFP-positive cells at 2 weeks posttransplantation. The regenerated nerves included Tuj1-positive axons, RECA1 and GFP double-positive blood vessels, and S100 and GFP double-positive Schwann-like supportive cells. Osmium-toluidine blue staining revealed that the regenerated nerves contained myelinated fibers. Moreover, fluorescent retrograde tracing analysis by application of Fluoro-Gold into the regenerated nerves demonstrated the presence of Fluoro-Gold-positive motor neurons in the facial nucleus of the rat brain. These results suggest that the transplanted dental pulp cells formed blood vessels and myelinating tissue and contributed to the promotion of normal nerve regeneration.     

Ⅵ～Ⅷ对脑神经生物塑化薄片断层解剖与MRI对照研究

目的 :对展神经、面神经、前庭蜗神经进行塑化切片与MRI对照研究 ,获得正常影像和断层解剖资料。方法 :采用生物塑化技术制作脑神经横断位 8例、矢状位和冠状位各 1例薄层切片 ,同时采用FLASH -3D序列完成头颅标本及 3 0例正常人脑干MR扫描 ,以MPR技术完整显示展神经、面神经、前庭蜗神经脑池段全程。结果 :塑化薄片断层、标本和活体MR扫描 ,脑神经行程、解剖形态均有良好的对应关系 ,Dorello管和展神经海绵窦段在塑化切片上显示良好 ,MRI仅能部分显示。结论 :生物塑化薄层切片能够对脑神经及相关结构进行准确显示 ,是脑神经影像学研究的重要对比方法     

枕小和耳大神经移植修复面神经缺损的应用解剖

目的为枕小和耳大神经移植修复面神经缺损提供形态学基础。方法在32侧成人标本上,观测颈丛皮神经的行程、长度、横径和血供情况。结果枕小和耳大神经的长度分别为（7．7±0．8）cm和（7．9±0．9）cm;横径分别为（1．6±0．4）mm和（2．2±0．4）mm。枕小和耳大神经的血供主要来源于颈升动脉神经支、枕动脉的胸锁乳突肌皮支和耳后动脉皮支。结论枕小和耳大神经可作为面神经移植的一个合适而易得的理想供体,行自体神经游离移植或吻合血管神经移植用以修复各种原因所致的面神经缺损。     

化学去细胞异体神经联合骨髓间充质干细胞修复损伤坐骨神经的生物力学评价

文题释义： 生物力学：是应用力学原理和方法对生物体中的力学问题定量研究的生物物理学分支,其研究范围从生物整体到系统、器官(包括血液、体液、脏器、骨骼等),从鸟飞、鱼游、鞭毛和纤毛运动到植物体液的输运等。生物力学的基础是能量守恒、动量定律、质量守恒定律并加上描写物性的本构方程。生物力学研究的重点是与生理学、医学有关的力学问题。依研究对象的不同可分为生物流体力学、生物固体力学和运动生物力学等。 周围神经研究热点和发展趋势：周围神经损伤的治疗已经取得了突破性的进展,但由于周围神经走行长、结构复杂、功能多样,损伤后其自身和靶器官会发生改变,而且是动态的、有时间效应的改变,因此以整体统一的理念研究周围神经损伤是今后研究重点。如何将基础研究与临床相结合,最终实现成果转化服务于患者,是研究者们的努力方向。国内外学者对自体神经损伤移植替代材料的研究从未间断过。随着组织工程和生物力学的发展,同种异体神经和人工神经已逐渐应用于临床,去细胞神经移植物也已取得了很好的临床效果。背景：以拉伸力学指标评估化学去细胞异体神经联合骨髓间充质干细胞治疗坐骨神经损伤的效果,目前鲜有报道。 目的：评估化学去细胞异体神经联合骨髓间充质干细胞移植治疗坐骨神经损伤的效果。 方法：将45只SD大鼠随机分为自体神经移植组、化学去细胞异体神经移植组、化学去细胞异体神经联合骨髓间充质干细胞移植组,每组15只,制备10 mm坐骨神经损伤模型,分别以自体神经、化学去细胞异体神经、化学去细胞异体神经联合骨髓间充质干细胞修复坐骨神经损伤,术后20周,进行坐骨神经电生理检测和坐骨神经单向拉伸实验。 结果与结论：①化学去细胞异体神经联合骨髓间充质干细胞移植组与自体神经移植组波幅值、运动传导速度值大于化学去细胞异体神经移植组(P < 0.05);②化学去细胞异体神经联合骨髓间充质干细胞移植组与自体神经移植组拉伸弹性限度应变、弹性限度应力、最大应变、最大应力大于化学去细胞异体神经移植组(P < 0.05);③结果表明,化学去细胞异体神经联合骨髓间充质干细胞移植对坐骨神经损伤具有明显的修复效果。ORCID: 0000-0002-1029-8117(吴志峰) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

Connections between the facial, vestibular and cochlear nerve bundles within the internal auditory canal

The vestibular, cochlear and facial nerves have a common course in the internal auditory canal (IAC). In this study we investigated the average number of nerve fibres, the average cross-sectional areas of the nerves and nerve fibres, and the apparent connections between the facial, cochlear and vestibular nerve bundles within the IAC, using light and scanning electron microscopy. The anatomical localization of the nerves within the IAC was not straightforward. The general course showed that the nerves rotated anticlockwise in the right ear from the inner ear end towards the brainstem end and vice versa for the left ear. The average number of fibres forming vestibular, cochlear, and facial nerves was not constant during their courses within the IAC. The superior and the inferior vestibular nerves showed an increase in the number of nerve fibres from the inner ear end towards the brainstem end of the IAC, whereas the facial and the cochlear nerves showed a reduction in the number of fibres. This suggests that some of the superior and inferior vestibular nerve bundles may receive fibres from the facial and/or cochlear nerves. Scanning electron microscopic evaluations showed superior vestibular-facial and inferior vestibular-cochlear connections within the IAC, but no facial-cochlear connections were observed. Connections between the nerves of the IAC can explain the unexpected vestibular disturbances in facial paralysis or persistence of tinnitus after cochlear neurectomy in intractable tinnitus cases. The present study offers morphometric and scanning electron microscopic data on the fibre connections of the nerves of the IAC.     

种植施万细胞的脱细胞同种异体神经移植物对大鼠坐骨神经缺损的修复作用

目的观察种植施万细胞的脱细胞同种异体神经移植物,桥接大鼠坐骨神经缺损后的神经再生。方法应用酶反复消化法与差速贴壁法体外分离培养乳鼠施万细胞;显微注射法将施万细胞种植到脱细胞同种异体神经移植物内;再应用种植施万细胞的脱细胞同种异体神经移植物桥接大鼠坐骨神经10 mm缺损。光镜、透射电镜和扫描电镜观察再生神经的形态结构、有髓神经纤维数量、平均髓鞘厚度并进行统计学分析。结果光、电镜观察到实验组(SCs+ARSN)的施万细胞在再生神经纤维中互相连结纵行排列成类似Büngner带样细胞链,对照组(ARSN)未见到施万细胞的链状排列。实验组再生有髓神经纤维的髓鞘厚度较对照组均匀且较厚,有髓神经纤维数量和平均髓鞘厚度明显多于对照组(P<0.05)。结论种植施万细胞的脱细胞同种异体神经移植物对缺损的坐骨神经再生有更加有效的促进作用。     

Correlation between MRI and operative findings in Bell's palsy and Ramsay Hunt syndrome

PURPOSE: To investigate the correlation between gadolinium enhanced magnetic resonance image (MRI) results and surgical findings of facial nerves in Bell's palsy and Ramsay Hunt syndrome. MATERIALS AND METHODS: From 1995 to 2004, MRI was performed on 13 patients with Bell's palsy or Ramsay Hunt syndrome, who were offered with surgical decompression of the facial nerve through the middle cranial fossa approach. Gadolinium enhanced MRI was performed on all patients and the enhancement of the facial nerve was evaluated by radiology specialists. Operative findings including the degree of the facial nerve segment swelling were examined. Furthermore, the time interval from the onset of palsy to surgery was evaluated. RESULTS: Swelling of facial nerve segments was found in patients with enhanced facial nerves from MRI. The swelling of the facial nerve in the labyrinthine segment in particular was identified in all patients with enhanced labyrinthine segments in MRI. The intraoperative swelling of geniculate ganglion of facial nerve was found in 78% of patients with enhanced facial segment in MRI (p=0.01). The intraoperative swelling of tympanic segment was observed from fourth to ninth weeks after the onset of palsy. CONCLUSION: MRI enhancement of facial nerves in Bell's palsy and Ramsay Hunt syndrome is associated with the extent of intratemporal lesions of facial nerves, especially in the labyrinthine segment.