与雪旺细胞共培养诱导脂肪来源干细胞向神经元样细胞分化的实验研究

目的探讨非接触共培养条件下,大鼠SCs对脂肪来源干细胞(adipose-derived stem cells,ADSCs)的诱导分化效应,为神经组织工程寻找理想种子细胞提供参考。方法取新生1~2 d SD大鼠双侧坐骨神经,酶消化法分离培养SCs,并行S-100免疫荧光染色鉴定。取成年雄性SD大鼠腹股沟和腹膜后脂肪组织,差速贴壁法纯化获得ADSCs并传代,流式细胞仪检测第3代细胞表型CD29、CD34、CD45、CD73、CD90和CD105。取原代SCs和第3代ADSCs按2∶1比例于Transwell共培养系统共培养(实验组),以单纯培养ADSCs作为对照组。倒置相差显微镜下观察细胞形态学改变,培养14 d流式细胞仪检测神经元特异性烯醇化酶(neuron-specific enolase,NSE)表达,免疫荧光染色观察微管相关蛋白2(microtubule-associated protein 2,MAP2)、神经元核蛋白(neuronal nuclei protein,Neu N)和胶质原纤维酸性蛋白(glial fibrillary acidic protein,GFAP)神经元特异性标志物表达情况,并计算阳性细胞率。结果成功分离培养ADSCs并能连续传代,流式细胞仪检测示其高表达CD29、CD90、CD73和CD105,低表达CD34和CD45。倒置相差显微镜下见,实验组共培养后原本呈梭形的ADSCs以胞核为中心收缩,胞体折光性增强,胞体伸出多个长而粗的突起;对照组ADSCs形态无显著变化。流式细胞仪及免疫荧光染色示,共培养后14 d实验组均表达神经元特异性标志物NSE、MAP2、Neu N、GFAP,且阳性细胞率显著高于对照组(P0.01)。结论 SCs与ADSCs非接触共培养,两者不仅能够共生,且SCs能显著促进ADSCs向神经元样细胞分化。     

周围神经损伤修复技术应用及组织工程技术的研究进展

据统计,全世界每年约一百多万人发生周围神经损害,多由严重创伤、肿瘤切除、先天性畸形等原因所致,周围神经缺损的修复与重建仍是当前临床神经领域的一大难题。现就周围神经损伤修复技术应用及组织工程研究进展综述如下。     

血管化纳米诱导骨的构建及促进兔横突间融合的研究

我们研制释放血管内皮生长因子(VEGF)的人工膜材,作为毛细血管网的基质材料,同步构筑血管化纳米诱导骨,并应用于兔横突间融合部位,观察其能否促进脊柱融合,分析其作用机制,探讨应用潜力.     

人脐血单个核细胞联合雪旺细胞修复脊髓损伤的实验研究

目的 探讨人脐血单个核细胞(haman cord blood mononuclear cells,HCMNCs)与大鼠雪旺细胞(Schwann cells,SCs)联合移植应用于大鼠脊髓损伤的疗效.方法 健康成年8周龄Wistar雌性大鼠40只,体重(200±30)g.利用Impactor Model Ⅱ型打击器制成T10脊髓损伤模型.40只大鼠随机分成4组,每组10只,即DMEM实验对照组、HCMNCs移植组、SCs移植组、HCMNCs+SCs联合移植组.用HE染色、顺行示踪染色及电镜观察脊髓损伤处轴突再生情况,对各组实验动物脊髓损伤后肢体功能的恢复情况进行行为学评分(BBB评分)及脚印分析实验,综合评估脊髓功能恢复程度.结果 HCMNCs+SCs联合移植治疗能够明显促进神经再生,改善功能,减少空洞形成.BBB评分和脚印分析实验显示各组疗效比较,差异有统计学意义.组织学和电镜观察神经轴突再生情况与功能学检查的结果 相吻合.结论 以分泌各种神经营养因子为特点的SCs为平台,联合移植后诱导HCMNCs向神经元及神经胶质细胞分化,可促进脊髓损伤的恢复.     

调控Nurr1表达影响神经干细胞向多巴胺能神经元分化的研究进展

帕金森病（PD）是一种进行性加重的慢性中枢神经系统功能失调所致一系列退行性病变,主要表现为中脑黑质多巴胺（DA）能神经元分泌减少引起的震颤、僵直、运动迟缓等运动障碍及非运动障碍。传统的药物疗法是在短期内补充DA,可缓解症状;其中左旋多巴是治疗PD的首选药物,但长期服用患者会出现精神症状、开关现象、剂末现象和晨僵少动等不良反应^[1]。最新研究^[2]显示,神经干细胞（NSC）移植术有望达到治疗PD运动障碍的目的。     

柴胡皂苷a影响大鼠急性脊髓损伤后机体炎症水平表达的实验研究

Objective: To investigate the effect of saikosaponina on the expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the early stage of acute spinal cord injury in rats. Methods: Thirty SD rats were randomly divided into sham operation group, simple injury group, saikosaponin a 5 mg / kg group, 10 mg / kg group, 20 mg / kg group, respectively. The acute spinal cord injury model was made by modified ^,Allen^,s heavy blow method. The rats in the injury group were injected with normal saline only, and the drug groups were intraperitoneally injected with the different dosages. All the animals were sacrificed at 24 h after the operation. The spinal cord and the vena cava sera of the injured segment were taken for 10 minutes and 2 minutes, respectively. The levels of TNF-αand IL-6 in the spinal cord tissue and serum were detected by enzyme-linked immunosorbent assay（ELISA）. Results: TNF-αand IL-6 in the spinal cord tissue and serum of the rats in the pure injury group were higher than those in the sham group (P <0.05). The levels of TNF-αand IL-6 in spinal cord tissue and serum of rats treated with saikosaponin a 5 mg / kg, 10 mg / kg and 20 mg / kg were lower than those of the injury group, and the difference was statistically significant (P < 0.05). The expressions of TNF-α and IL-6 in the spinal cord tissue of rats treated with 10 mg / group and 20 mg / kg group the difference were not statistically significant (P> 0.05). The levels of TNF-α and IL-6 in serum of rats treated with 10mg / kg and 20 mg/kg were lower than those of 5mg/kg group (P> 0.05). Conclusion: Saikosaponina could reduce the secondary immune inflammatory response by affecting the early expression of inflammatory factors to improve the body’s immune function in acute spinal cord injury. The10mg/kg group is superior to the other groups, and the drug may partially pass through the spinal blood barrier.     

力学调控长链非编码RNA对间充质干细胞成骨分化作用的研究进展

随着第3代高通量测序技术和组织工程的发展,近年来研究发现,许多长链非编码RNA（long-chain non-coding RNAs,LncRNAs）在力学调节间充质干细胞（mesenchymal stem cells,MSCs）的成骨分化中起重要作用。力学因素调控相关的LncRNAs,然后LncRNAs通过转录、转录后以及表观遗传等多层次发挥调节作用,进一步调控骨相关细胞功能。LncRNAs也可能作为力学响应分子参与MSCs成骨分化、骨重建以及骨骼系统疾病的病理过程。     

力学过载对成骨细胞损伤及中药修复的研究进展

骨的生长代谢由成骨细胞的成骨及破骨细胞的吸收共同控制,而成骨细胞在成骨过程中起主要作用。过载影响成骨细胞的增殖与分化,其加载方式、强度、持续时间等因素都可改变成骨细胞的生物特性,进而影响成骨细胞的功能活性。而过载引起成骨细胞应答的机制还处于探索阶段,需要进一步深入研究。大量实验证明,中药淫羊藿苷促进成骨细胞增殖及分化,一定浓度的淫羊藿苷对骨损伤修复起重要作用。综述骨细胞对过载刺激后的反应及淫羊藿苷对成骨细胞的损伤修复。     

IL-11调控 gp130/JAK2/STAT3信号通路促进Wistar大鼠雪旺细胞中PMP22的表达

目的:探索白介素-11对大鼠雪旺细胞分泌髓鞘蛋白的调节作用及相关机制。方法:取成年Wistar大鼠坐骨神经,分离纯化培养雪旺细胞。加入重组大鼠白介素-11,分别采用realtime RT-qPCR和Western boltting检测雪旺细胞髓鞘蛋白的表达情况,进一步检测IL-11调控髓鞘蛋白表达的相关通路的激活情况。结果:Realtime RT-qPCR和Western boltting结果均提示白介素-11可以显著提高雪旺细胞中周围神经髓鞘蛋白22的表达水平,而对雪旺细胞中髓鞘碱性蛋白和髓鞘蛋白零的表达水平则没无明显影响,该生理作用可被JAK特异性抑制剂AG490所抑制。在经过IL-11处理的SCs 细胞中可以观察到pJAK2（Y1007＋Y1008）和pSTAT3（Y705）含量明显高于对照组且伴随着SCs中gp130表达的上调。结论:白介素-11可通过调剂gp30/JAK2/STAT3信号通路以促进雪旺细胞中周围神经髓鞘蛋白22的表达。     

胶原/丝素蛋白神经导管修复周围神经缺损的研究与应用进展

BACKGROUND: Peripheral nerve defect due to limb dysfunction has always been the difficulty faced by the medical profession. Ideal materials and processing technology for constructing a tissue engineering scaffold targeting peripheral nerve repair are still in research stage. OBJECTIVE: To review the research progress in peripheral nerve repair using collagen/silk fibroin nerve conduits. METHODS:In this paper, the first author retrieved the PubMed and CNKI from 2003 to 2016 to search articles regarding methods of constructing artificial nerve scaffolds and selection of raw materials. Data from these articles were collected, summarized and analyzed. RESULTS AND CONCLUSION: Forty-six articles were included for final analysis. Collagen and its degradation products trigger no inflammatory response in the host because of high biocompatibility and biodegradability. However, its use is largely limited by its rapid degradation and poor physical performance. Silk fibroin has a high flexibility and biocompatibility, and exhibits a slow degradation in vivo. As a rapid prototyping technique, three-dimensional printing can print various forms of scaffolds within a short time, characterized as high-quality pore structure and large-scale production. Given these, the collagen/silk fibroin nerve conduit prepared using the three-dimensional printing technology can maintain the biocompatibility and even improve the mechanical properties of the raw materials. Until now, more investigations on nerve repair using collagen or silk fibroin have been done, and we have never stopped improving the production process of these scaffolds. Therefore, the collagen/silk fibroin scaffold prepared using the three-dimensional printing technology is expected to become the main candidate for the repair of peripheral nerve defects.