NGF联合自体骨髓干细胞动员治疗重型颅脑损伤疗效观察

目的探讨神经生长因子(NGF)联合自体骨髓干细胞(BMSCs)动员治疗重型颅脑损伤的临床疗效和安全性。方法选取60例重型颅脑损伤患者按随机数字表分为对照组(30例,给予NGF和综合康复治疗)和观察组(30例,在对照组治疗基础上加用自体BMSCs动员治疗),比较2组患者神经功能缺损评分(NIHSS)、格拉斯哥预后评分(GOS)及不良反应。结果 2组患者治疗28d的NIHSS评分均较治疗前明显降低(P<0.05),且观察组较对照组降低更为显著(P<0.05);观察组伤后6个月GOS评分明显高于对照组(P<0.05),外周血CD34+、CD133+占外周血单核细胞比率治疗3周时明显高于对照组(P<0.05)。2组患者均未出现明显不良反应。结论 NGF联合自体BMSCs动员及综合康复治疗可促进重型颅脑损伤患者神经损伤的修复,显著改善患者神经功能。     

骨髓间充质干细胞移植治疗大鼠脑内出血

目的研究自体骨髓间充质干细胞(BMSCs)移植对脑出血大鼠的运动功能改善效果,以及采用不同移植方法后细胞在受损脑组织中的分布规律。方法采用立体定向技术向纹状体内注射自体新鲜心室腔血液构建SD大鼠脑出血模型,30只雄性SD大鼠随机分为A组:自然恢复组(对照组);B组:尾静脉移植组;C组:立体定向移植组,于移植后7 d、14 d进行神经功能缺损评分,并在荧光显微镜下观察BMSCs存活、迁移和分布情况。结果 C组大鼠神经功能恢复优于B组,BMSCs均能够迁移分布至大鼠脑损伤区。结论 BMSCs移植对脑出血后大鼠神经功能的恢复具有明显的促进作用。立体定向移植组大鼠神经功能恢复优于尾静脉移植组。     

神经生长因子基因修饰骨髓间充质干细胞移植治疗脑梗死大鼠的实验研究

目的观察神经生长因子基因(NGF)修饰的骨髓间充质干细胞(BMSCs)移植治疗脑梗死的作用及可能机制。方法采用线栓法制备大鼠脑梗死模型,将符合条件的30只脑梗死模型大鼠按随机数字表法分为模型组(n=10),BMSCs移植组(n=10)和NGF-BMSCs移植组(n=10),分别经尾静脉注射PBS、BrdU标记的BMSCs和NGF-BMSCs各1mL。分别于术后1d、7d和14d采用改良神经功能损害评分(mNSS)对各组大鼠进行神经功能评估,于术后14d应用HE染色观察脑组织病理情况,应用免疫荧光组织化学检测BrdU标记的移植细胞存活状况和TUNEL法检测脑组织中细胞凋亡情况。结果NGFBMSCs组和BMSCs组mNSS评分和TUNEL阳性细胞数较Model组减低(P0.05),且NGF-BMSCs组较BMSCs组更低(P0.05);HE染色显示NGF-BMSCs组和BMSCs组较Model组脑组织损伤及细胞丢失较轻,NGF-BMSCs组更明显;并且NGF-BMSCs组中的BrdU阳性细胞数较BMSCs组增多(P0.05)。结论 NGF基因修饰的BMSCs移植较单纯BMSCs移植能进一步改善脑梗死大鼠的神经功能,其机制为能够促进植入的BMSCs在脑内存活和减轻神经细胞凋亡。     

注射用鼠神经生长因子治疗急性脑出血62例疗效观察

目的观察注射用鼠神经生长因子治疗急性脑出血的临床疗效。方法将急性脑出血患者62例分为2组。观察组31例,应用注射用鼠神经生长因子9 000AU,溶于生理盐水2mL,肌内注射,1次/d,治疗4周;对照组31例,应用胞二磷胆碱注射液,0.5g,静滴,1次/d,连续治疗4周。治疗开始前及治疗4周后,对2组患者分别进行神经功能缺损评分及日常生活活动量评分,并对比两者差别。结果注射用鼠神经生长因子观察组与对照组相比治疗后神经功能缺损程度评分、日常生活活动量评分差异均有统计学意义(P<0.05)。结论注射用鼠神经生长因子对脑出血患者有明显的神经保护作用,临床疗效可靠。     

自体骨髓间充质干细胞移植辅助高压氧治疗高血压脑出血临床观察

目的：观察自体骨髓间充质干细胞移植辅助高压氧治疗高血压脑出血的疗效。方法将102例高血压脑出血患者随机分为治疗组（自体骨髓间充质干细胞移植＋高压氧治疗组）和对照组（高压氧治疗组）,比较2组患者治疗前和治疗后6个月的神经功能缺损程度评分（NIHSS）和日常生活活动能力评分（Barthel指数）,治疗后6个月格拉斯哥预后评分（GOS）及远期生活质量评估量表评分（KPS）。结果治疗后6个月与对照组比较,NIHSS评分均有显著降低Barthel指数均有显著升高（P＜0·05）,治疗组GOS及KPS评分显著好于对照组（P＜0·05）。结论自体骨髓间充质干细胞移植辅助高压氧治疗能显著促进高血压脑出血患者神经功能恢复,提高患者生存率和生活质量。     

骨髓基质细胞及血管内皮祖细胞共移植对局灶性脑缺血大鼠系列生长因子表达的影响

目的 观察经静脉移植骨髓基质细胞(BMSCs)及血管内皮祖细胞(EPCs)后血管内皮生长因子(VEGF)、基质细胞衍生因子-1(SDF-1)、碱性成纤维生长因子(bFGF)、胰岛素样生长因子-1(IGF-1)、转化生长因子β(TGF-β)、血小板源性生长因子(PDGF)、脑源性生长因子(BDNF)、胶质源性生长因子(GDNF)和神经生长因子(NGF)这9种细胞因子的表达. 方法 80只健康成年Wistar 大鼠按照随机数字表法分为模型对照组、BMSCs移植组、EPCs移植组及联合移植组,每组20只.线栓法建立大鼠大脑中动脉闭塞模型.造模24h后,分别取浓度为3x106个/mL的BMSCs、EPCs悬液及两者混合液1mL经鼠尾静脉移植入BMSCs移植组、EPCs移植组及联合移植组;模型对照组鼠尾静脉注射1 mL生理盐水.移植后7d,实时荧光PCR、Western boltting检测各组大鼠脑内生长因子的表达. 结果 联合移植组中bFGF、VEGF、BDNF mRNA表达水平最高,与其他3组比较差异均有统计学意义(P＜0.05).BMSCs移植组中NGF、GDNF、TGF-β mRNA表达水平最高,与其他3组比较差异均有统计学意义(P＜0.05),联合移植组次之.EPCs组中PDGF-BB、IGF-1、SDF-1 mRNA表达水平最高,与其他3组比较差异均有统计学意义(P＜0.05),联合移植组次之. 结论 BMSCs及EPCs联合静脉移植可上调脑内生长因子表达水平,为提高细胞移植效率提供了新的尝试途径.     

NGF基因转染骨髓间充质干细胞治疗脑出血的分析

目的 研究NGF基因转染骨髓间充质干细胞(BMSCs)对脑出血大鼠运动功能的改善效果.方法 制作Wistar大鼠尾状核脑出血模型,随机分为A组:脑出血模型组;B组:细胞培养液移植组;C组:BMSCs移植组;D组:NGF基因修饰的BMSCs移植组,4组于移植后1、7、14、28 d进行神经功能缺损评分,并经免疫组织化学鉴定了解移植细胞在大鼠脑内的生存及分化情况.结果 C、D 2组的大鼠运动功能改善好于A、B组,并且D组改善又好于C组,免疫组化显示移植细胞生长良好,并且在血肿灶周围聚集融合,免疫荧光双标证实被移植细胞在脑内分化为神经元、星型胶质细胞.结论 NGF基因修饰的BMSCs移植治疗能显著改善脑出血动物的运动功能,是一种很有发展前途的方法,值得进行更深入的研究.     

自体骨髓间充质干细胞移植治疗脑出血的疗效观察

目的探讨自体骨髓间充质干细胞治疗脑出血近期有效性和安全性。方法 2009-01—2012-01我院神经外科收治的脑出血患者45例。移植组20例通过蛛网膜下腔注射方式行自体骨髓间充质干细胞移植,选择同期入院但未行干细胞移植的25例作为对照组。分别于移植前和移植后6个月进行神经功能缺损程度评定(NIHSS评分)和日常生活活动能力评定(Barthel指数)。同期随访血常规、生化全项、肿瘤标记物以观察其安全性。结果移植前2组NIHSS评分及Barthel指数基本相似。与对照组比较,移植后6个月治疗组NIHSS评分显著降低(P<0.05),Barthel指数显著升高(P<0.05)。与移植前比较,治疗组在移植后6个月NIHSS评分显著降低(P<0.05),BaRhel指数显著升高(P<0.05)。2例患者在移植后出现一过性发热,予对症处理后症状缓解。随访各项血液检查结果无明显异常。结论采用自体骨髓间充质干细胞治疗脑出血临床疗效安全有效,近期临床疗效明显,远期治疗效果有待观察。     

自体骨髓干细胞移植联合神经营养因子治疗帕金森病的疗效

目的：初步探讨自体骨髓干细胞（bone marrow stem cell ,BMSCs）动员移植联合神经营养因子治疗帕金森病的安全性及疗效。方法将106例 PD患者随即分为对照组与观察组,对照组在口服美多巴基础上给予鼠神经生长因子（mNGF）肌内注射,1次/d ,连续4周;观察组在对照组治疗基础上行BMSCs动员移植治疗,6个月后采用 Hoehn-Yahr分期评分、UPDRS评分、改良Webster量表评定2组疗效。结果 BMSCs动员移植联合神经营养因子在改善患者运动症状及平衡障碍、日常活动及行为,精神、情绪等非运动症状方面较单用神经营养因子疗效更佳。结论自体骨髓干细胞动员移植联合神经营养因子治疗帕金森病安全,疗效满意。     

自体骨髓间充质干细胞移植治疗颅脑创伤后神经功能障碍的护理

目的：探索自体骨髓间充质干细胞移植治疗颅脑创伤后神经功能障碍的护理措施。方法选取颅脑创伤后神经功能障碍患者76例,分别实施常规治疗和自体骨髓间充质干细胞移植治疗,通过对比2组患者治疗前、治疗后6个月神经功能缺损程度（NIHSS）和日常生活能力评分（Barthel指数）,评价治疗和护理效果。结果所有患者均治疗成功出院,移植组较对照组术后6个月NIHSS评分显著降低（P＜0.001）,Barthel指数显著升高（P＜0.001）。结论自体骨髓间充质干细胞移植治疗能促进患者神经功能恢复,提高患者生活能力,治疗前心理护理,移植后严密观察病情变化、不良反应,持之以恒的功能康复计划是促进患者康复的重要保障。     

脐血单个核细胞对血管性痴呆大鼠脑组织BDNF、NGF的影响

目的 观察颈内动脉输注脐血单个核细胞(HCMNCs)对血管性痴呆(VD)大鼠脑组织脑源性神经营养因子(BDNF)和神经生长因子(NGF)表达及大鼠学习记忆能力的影响. 方法 改良Pulsinellis四血管阻断法建立VD大鼠模型;大鼠受用随机数字表法分为对照组、模型组和治疗组;每组又分为2,4,8周三个时相点,每时相点12只.体外分离HCMNCs,术后24h颈内动脉输注3x106个BrdU标记细胞于治疗组;利用穿梭箱系统和ELISA法检测注射HCMNCs后2,4,8周VD大鼠学习记忆能力以及脑组织BDNF和NGF含量的变化. 结果 模型组大鼠主动回避反应比率显著低于对照组(P＜0.01),治疗组较模型组显著提高(P＜0.01).术后2周模型组大鼠脑BDNF、NGF含量较对照组显著增高(P＜0.01),4周时达到高峰(P＜0.01),8周时则明显下降,与2,4周时相比有显著性差异(P＜0.05);治疗组大鼠脑BDNF、NGF含量较模型组显著升高(P＜0.01),4周时最高(P＜0.05,P＜0.01),8周时略有下降,但仍维持在较高水平,与2,4周时相比无统计学意义(P＞0.05).结论 颈内动脉输注HCMNCs可显著改善VD大鼠学习记忆能力,推测原因为通过增加VD大鼠脑组织BDNF和NGF发挥作用.     

骨髓间质干细胞移植减轻犬急性脑梗死模型的缺血性脑损伤研究

目的观察骨髓间质干细胞(BMSCs)对犬急性缺血脑组织的保护作用,并探讨其可能的机制。方法将24只成年杂交犬随机分为治疗组及对照组,DSA引导下行自体血栓栓塞大脑中动脉闭塞缺血模型制作,并抽取骨髓提取BMSCs,传代并给予4’,6-二脒基-2-苯基吲哚(DAPI)标记;1周后开颅行多点脑内注射BMSCs移植;移植后1周行脑DWI序列扫描,计算梗死灶体积;4周后将犬处死,每组随机选择6只动物取脑标本行TTC染色测定梗死灶体积;另外6只动物进行HE染色、VG染色、TUNEL染色评价脑损伤情况;免疫荧光染色了解脑源性神经营养因子(BDNF)、碱性成纤维生长因子(b FGF)、胰岛素样生长因子1(IGF-1)和血管内皮生长因子(VEGF)的表达情况。结果治疗组梗死灶内广泛存在DAPI阳性细胞。治疗组的梗死灶体积明显小于对照组,P0.01。治疗组梗死灶范围、梗死灶内细胞坏死、胶质增生和胶质瘢痕均较对照组减轻。治疗组凋亡细胞明显少于对照组,P0.05。治疗组细胞因子BDNF、BFGF、IGF-1和VEGF表达阳性的细胞均显著多于对照组。结论脑梗死后给予BMSCs移植,BMSCs能存活并自行向梗死灶迁移,并发挥脑保护作用,其机制可能和分泌多种神经营养因子有关。     

Bone marrow mesenchymal stem cells stimulated with low‐intensity pulsed ultrasound: Better choice of transplantation treatment for spinal cord injury

Stem cell transplantation, especially treatment with bone marrow mesenchymal stem cells (BMSCs), has been considered a promising therapy for the locomotor and neurological recovery of spinal cord injury (SCI) patients. However, the clinical benefits of BMSCs transplantation remain limited because of the considerably low viability and inhibitory microenvironment. In our research, low‐intensity pulsed ultrasound (LIPUS), which has been widely applied to clinical applications and fundamental research, was employed to improve the properties of BMSCs. The most suitable intensity of LIPUS stimulation was determined. Furthermore, the optimized BMSCs were transplanted into the epicenter of injured spinal cord in rats, which were randomized into four groups: (a) Sham group (n = 10), rats received laminectomy only and the spinal cord remained intact. (b) Injury group (n = 10), rats with contused spinal cord subjected to the microinjection of PBS solution. (c) BMSCs transplantation group (n = 10), rats with contused spinal cord were injected with BMSCs without any priming. (d) LIPUS‐BMSCs transplantation group (n = 10), BMSCs stimulated with LIPUS were injected at the injured epicenter after contusion. Rats were then subjected to behavioral tests, immunohistochemistry, and histological observation. It was found that BMSCs stimulated with LIPUS obtained higher cell viability, migration, and neurotrophic factors expression in vitro. The rate of apoptosis remained constant. After transplantation of BMSCs and LIPUS‐BMSCs postinjury, locomotor function was significantly improved in LIPUS‐BMSCs transplantation group with higher level of brain‐derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in the epicenter, and the expression of neurotrophic receptor was also enhanced. Histological observation demonstrated reduced cavity formation in LIPUS‐BMSCs transplantation group when comparing with other groups. The results suggested LIPUS can improve BMSCs viability and neurotrophic factors expression in vitro, and transplantation of LIPUS‐BMSCs could promote better functional recovery, indicating possible clinical application for the treatment of SCI.     

Sciatic nerve repair by acellular nerve xenografts implanted with BMSCs in rats xenograft combined with BMSCs

Acellular nerves possess the structural and biochemical features similar to those of naive endoneurial tubes, and have been proved bioactive for allogeneil graft in nerve tissue engineering. However, the source of allogenic donators is restricted in clinical treatment. To explore sufficient substitutes for acellular nerve allografts (ANA), we investigated the effectiveness of acellular nerve xenografts (ANX) combined with bone marrow stromal cells (BMSCs) on repairing peripheral nerve injuries. The acellular nerves derived from Sprague-Dawley rats and New Zealand rabbits were prepared, respectively, and BMSCs were implanted into the nerve scaffolds and cultured in vitro. All the grafts were employed to bridge 1 cm rat sciatic nerve gaps. Fifty Wistar rats were randomly divided into five groups (n = 10 per group): ANA group, ANX group, BMSCs-laden ANA group, BMSCs-laden ANX group, and autologous nerve graft group. At 8 weeks post-transplantation, electrophysiological study was performed and the regenerated nerves were assayed morphologically. Besides, growth-promoting factors in the regenerated tissues following the BMSCs integration were detected. The results indicated that compared with the acellular nerve control groups, nerve regeneration and functional rehabilitation for the xenogenic nerve transplantation integrated with BMSCs were advanced significantly, and the rehabilitation efficacy was comparable with that of the autografting. The expression of neurotrophic factors in the regenerated nerves, together with that of brain-derived neurotrophic factor (BDNF) in the spinal cord and muscles were elevated largely. In conclusion, ANX implanted with BMSCs could replace allografts to promote nerve regeneration effectively, which offers a reliable approach for repairing peripheral nerve defects.     

骨髓基质细胞及血管内皮祖细胞移植对局灶性脑缺血大鼠神经功能恢复的影响

目的 观察经静脉移植骨髓基质细胞(BMSCs)及血管内皮祖细胞(EPCs)后,缺血性脑损伤大鼠神经系统功能恢复情况.方法 40只健康成年Wistar大鼠按随机数字表法分为模型组、移植BMSCs组、移植EPCS组、移植BMSCs/EPCs组,每组10只.线栓法建立大鼠大脑中动脉闭塞(MCAO)模型.造模后24h取1 mLBMSCs、EPCs、BMSCs/EPCs细胞悬液(3×10~6个/mL)分别经尾静脉注射移植入后3组大鼠.模型组大鼠注射等量生理盐水.移植前、移植后1、7、14、28 d,采用旋转试验、躯体感觉试验、神经系统功能评分(NSS)评估各组大鼠神经功能的恢复情况.移植后28d,免疫荧光染色检测各组大鼠缺血脑组织的微血管密度(MVD).结果 移植后第7天,旋转试验显示移植BMSCs/EPCs组大鼠旋转时间长于其他3组,躯体感觉试验和NSS评分分别显示移植BMSCs/EPCs组大鼠移物时间和NSS评分低于其他3组,差异有统计学意义(P<0.05);移植后28 d,免疫荧光染色检测结果显示缺血脑组织MVD明显高于其他3组,差异有统计学意义(P<0.05).结论 静脉BMSCs/EPCS联合移植可增强缺血性脑损伤功能的恢复.     

氧化损伤星形胶质细胞与骨髓基质细胞共培养的实验研究

目的 观察氧化损伤星形胶质细胞与骨髓基质细胞(BMSCs)共培养时细胞损伤恢复及存活能力的改变,以进一步探讨BMSCs修复脑损伤的机制.方法 过氧化氢(H2O2)损伤体外培养的星形胶质细胞,与BMSCs共培养后光镜下观察细胞生存情况.分别在培养后第1、3天取细胞培养液应用EuSA试剂盒测定碱性成纤维细胞生长因子(bFGF)、脑源性神经营养因子(BDNF)、神经生长因子(NGF)以及表皮生长因子(EGF)的分泌水平,并与氧化损伤星形胶质细胞单独培养组和BMSCs单独培养组进行比较.结果 培养后3 d和7 d,共培养组星形胶质细胞的恢复及两种细胞的存活情况明显优于单独培养组.培养后1 d和3 d,共培养组细胞培养液中NGF、BDNF以及bFGF的浓度明显高于两个单独培养组之和,但是共培养组培养液中EGF的浓度明显低于两个单独培养组之和.结论 氧化损伤星形胶质细胞和BMSCs共培养能够提高星形胶质细胞的存活能力,增加NGF、BDNF以及bFGF的分泌水平.BMSCs移植修复脑损伤的功能可能部分是通过促进损伤星形胶质细胞恢复、减少星形胶质细胞死亡、增加神经营养和生长因子分泌完成.     

Transplantation of bone marrow stromal cells for peripheral nerve repair

Cell transplantation using bone marrow stromal cells (BMSCs) to alleviate neurological deficits has recently become the focus of research in regenerative medicine. Evidence suggests that secretion of various growth-promoting substances likely plays an important role in functional recovery against neurological diseases. In an attempt to identify a possible mechanism underlying the regenerative potential of BMSCs, this study investigated the production and possible contribution of neurotrophic factors by transected sciatic nerve defect in a rat model with a 15 mm gap. Cultured BMSCs became morphologically homogeneous with fibroblast-like shape after ex vivo expansion. We provided several pieces of evidence for the beneficial effects of implanted fibroblast-like BMSCs on sciatic nerve regeneration. When compared to silicone tube control animals, this treatment led to (i) improved walking behavior as measured by footprint analysis, (ii) reduced loss of gastrocnemius muscle weight and EMG magnitude, and (iii) greater number of regenerating axons within the tube. Cultured fibroblast-like BMSCs constitutively expressed trophic factors and supporting substances, including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), ciliary neurotrophic factor (CNTF), collagen, fibronectin, and laminin. The progression of the regenerative process after BMSC implantation was accompanied by elevated expression of neurotrophic factors at both early and later phases. These results taken together, in addition to documented Schwann cell-like differentiation, provide evidence indicating the strong association of neurotrophic factor production and the regenerative potential of implanted BMSCs.     

支气管哮喘大鼠肺内神经营养因子对气道高反应性影响的研究

目的 探讨支气管哮喘大鼠肺内神经营养因子水平的变化,及其与气道神经可塑性改变和气道高反应性间的关系. 方法 雄性SD大鼠40只按随机数字表法分为对照组、哮喘组、神经生长因子(NGF)+脑源性神经营养因子(BDNF)干预组和anti-NGF+anti-BDNF干预组(每组各10只),后三组通过鸡卵蛋白致敏激发建立哮喘模型并给予相应的干预.造模后第8周进行气道阻力测定,并留取左肺行HE染色及免疫组化染色检测肺组织中神经营养因子如NGF、BDNF的表达水平,RT-PCR法检测气道突触囊泡素(SYN)mRNA和神经丝(NF)mRNA的表达水平. 结果 哮喘组及NGF+BDNF干预组大鼠肺组织内可见大量炎症细胞浸润.哮喘组大鼠肺组织中NGF及BDNF的表达水平均明显高于对照组及anti-NGF+anti-BDNF干预组,差异有统计学意义(P＜0.05),而NGF+BDNF干预组肺组织中NGF及BDNF的表达水平明显高于哮喘组,差异亦有统计学意义 (P＜0.05).哮喘组及NGF+BDNF干预组较对照组的气道反应性增高,气道感觉神经元SYN mRNA及NF mRNA的表达水平也明显增高,差异均有统计学意义(P＜0.05).哮喘组大鼠肺组织中NGF及BDNF浓度与气道SYN表达呈正相关关系(r=0.889,P＜0.05;r=0.985,P＜0.05),与NF表达呈正相关关系(r=0.956,P＜0.05;r=0.927,P＜0.05),与气道反应性也呈正相关关系(r=0.938,P＜0.05;r=0.906,P＜0.05). 结论 NGF及BDNF可能参与了支气管哮喘大鼠气道神经可塑性改变的调节过程,从而导致气道高反应性的发生.     

HRP逆行示踪评价复合骨髓间充质干细胞的无细胞神经移植物

背景：作者前期将无细胞神经移植物与骨髓间充质干细胞复合培养,成功构建了组织工程人工神经。 目的：应用辣根过氧化物酶(HRP)神经逆行示踪技术对无细胞神经移植物复合骨髓间充质干细胞构建的神经移植复合体桥接大鼠坐骨神经缺损后运动神经元的保护作用进行评价。 方法：成年清洁级健康雄性SD大鼠,随机分成3组：①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。②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。③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。术后12周应用辣根过氧化物酶神经逆行示踪技术对脊髓前角运动神经元的再生进行评价。 结果与结论：术后12周脊髓前角运动神经元再生评价结果显示：实验组优于无细胞神经移植物组,而与自体神经移植物组相比差异无显著性意义。证实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,对大鼠脊髓运动神经元具有保护作用,可能达到与自体神经移植相似的效果。 关键词：无细胞神经移植物;骨髓间充质干细胞;辣根过氧化物酶;神经移植;大鼠     

Bone marrow stromal cells promote neurite outgrowth of spinal motor neurons by means of neurotrophic factors in vitro

Transplantation of bone marrow stromal cells (BMSCs) into spinal cord injury models has shown significant neural function recovery; however, the underlying mechanisms have not been fully understood. In the present study we examined the effect of BMSCs on neurite outgrowth of spinal motor neuron using an in vitro co-culture system. The ventral horn of the spinal grey matter was harvested from neonatal Sprague–Dawley rats, cultured with BMSCs, and immunostained for neurofilament-200 (NF-200). Neurite outgrowth of spinal motor neurons was measured using Image J software. ELISA was used to quantify neurotrophic factors such as brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) in culture media, and antibodies or exogenous neurotrophic factors were used to block or mimic the effect of BMSCs on neurite outgrowth, respectively. The results showed that neurite outgrowth significantly increased in spinal motor neurons after co-cultured with BMSCs, while the secretion level of BDNF, GDNF and NGF was dramatically elevated in co-culture. However, the neurite outgrowth-promoting effect of BMSCs was found to significantly reduced using antibodies to BDNF, GDNF and NGF. In addition, a fraction of BMSCs was found to exhibit NF-200 immunoreactivity. These results indicated that BMSCs could promote neurite outgrowth of motor neurons by means of neurotrophic factors. The findings of the present study provided new cues for the treatment of spinal cord injury.