脱细胞猪角膜基质支架的制备及其生物相容性

BACKGROUND: Studies have found that a variety of biological materials can be used for preparing corneal stroma scaffolds that have good biocompatibility, but research on preparation and biocompatibility of the acellular porcine corneal stroma scaffold is little. OBJECTIVE: To explore the preparation and biocompatibility of the acellular porcine corneal stroma scaffold. METHODS: Acellular porcine corneal stroma scaffold and its extract were prepared. Well-grown human corneal stromal cells were selected and cultured in the extract of acellular porcine corneal stroma scaffold (experimental group) or in the complete medium (control group), respectively. After 1, 2 and 3 days of culture, the proliferation ability of human corneal stromal cells was detected by MTT assay. In the meanwhile, human corneal cells were directly seeded onto the acellular porcine corneal stroma scaffold, and then the cell growth on the scaffold was detected using immunochemical method. RESULTS AND CONCLUSION: The number of human corneal stromal cells was in a rise with time in the two groups, and absorbance values had no significant difference between two groups at different time points of culture. Human corneal stromal cells grew well on the scaffold, and were positive for cell integrin β1, vimentin, aldehyde dehydrogenase 3A1, as well as CD34, CDK2 and K-Ras. These results show that the acellular porcine corneal stroma scaffold has no cytotoxicity, and has good biocompatibility.     

新型脱细胞半月板细胞外基质的制备及其生物相容性的研究

目的探索新型脱细胞半月板细胞外基质(d MECM)的制备方法,并对其细胞相容性进行研究。方法无菌条件下收集新鲜的猪半月板组织,切碎,采用湿法粉碎、差速离心的方法制备dM ECM生物材料。通过天狼猩红、甲苯胺蓝染色方法分别比较天然半月板与dM ECM中胶原以及糖胺聚糖含量的区别;采用Hoechst 33258荧光染色法观察脱细胞后dM ECM中DNA残留情况。将P3代的兔内侧半月板细胞种植在铺有dM ECM的盖玻片上,体外培养7 d后行扫描电镜检测其细胞相容性。结果本实验制备的dM ECM天狼猩红、甲苯胺蓝染色均呈阳性,并且与天然半月板染色类似;dM ECM行Hoechst33258染色呈阴性;扫描电镜结果显示种植于d MECM表面上的半月板细胞黏附紧密,可见大量细胞ECM的分泌。结论本实验制备的dM ECM可以很好地保留天然半月板ECM成分,有效地去除DNA物质,并且具有良好的生物相容性,是未来半月板组织工程领域非常有前景的支架材料。     

牛肌腱冻干脱细胞支架的生物力学特性

背景：目前的脱细胞方法在去除细胞的同时对细胞外基质存在一定的损伤,降低了脱细胞支架的生物力学性能。 目的：分析冻干牛肌腱脱细胞支架的生物力学特性。 方法：取新鲜小牛趾伸屈肌腱,去除小牛肌腱表面的滑膜、腱膜及软组织,双蒸水冲洗干净后低压冻干,通过物理方法制备肌腱纤维束60个,随机均分为两组,实验组于无菌操作下置入丝氨酸蛋白酶抑制剂,室温下持续24 h,无菌PBS冲洗后,再移入低浓度胰酶+乙醇混合溶液中,在不破坏细胞外基质的情况下去除细胞壁,室温下持续5 h,再将纤维束移入脱氧核糖核酸酶溶液中持续5 h,最后将已完成脱细胞步骤的支架使用PBS冲洗48 h,无菌室内室温下干燥;对照组不做处置。检测两组材料的弹性模量、耐久性及最大应力。 结果与结论：两组耐久性相似,但实验组在相同位移处的应力小于对照组;两组弹性模量比较差异无显著性意义,但实验组最大应力低于对照组(P < 0.01)。说明冻干脱细胞支架能够在一定程度上模仿牛肌腱的生物力学功能。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     

脱细胞异体真皮基质与人脂肪干细胞的生物相容性

背景：脱细胞异体真皮基质具有优良的生物相容性和组织细胞诱导功能。 目的：评价人脂肪干细胞与脱细胞异体真皮基质的生物相容性。 方法：取健康成年人吸脂术后的脂肪组织分离脂肪干细胞,并行原代与传代培养,传至第3代,将细胞与脱细胞异体真皮基质联合体外培养3,7 d,倒置相差显微镜和扫描电镜观察细胞在支架材料上的黏附、生长及增殖情况,并计算细胞在材料上的黏附率;XTT比色法检测细胞的生长增殖情况。 结果与结论：脂肪干细胞在支架材料上分布均匀,24 h内细胞开始伸展、黏附,二三天完全伸展变形,以梭形为主,呈网状排列;随着培养时间延长,支架上的细胞逐渐增多;人脂肪干细胞细胞与脱细胞异体真皮基质混合培养后平均黏附率为95.03%,并保持正常的生长增殖速度,表明支架对细胞具有良好的黏附性;脱细胞异体真皮基质材料与人脂肪干细胞复合后相容性良好。     

基因修饰牙龈成纤维细胞及脱细胞真皮基质制备牙周组织工程复合物

背景：前期研究发现人血小板源性生长因子B基因转染的牙龈成纤维细胞能够在体外快速增殖,且能够向胞外分泌血小板源性生长因子BB蛋白。目的：了解人血小板源性生长因子B基因修饰牙龈成纤维细胞植入脱细胞真皮基质后,在体内形成牙周组织工程化复合物的能力。方法：将人血小板源性生长因子B基因转染与未转染的Beagle犬牙龈成纤维细胞分别接种于脱细胞真皮基质上,观察细胞在脱细胞真皮基质上的生长情况。将人血小板源性生长因子B基因转染犬牙龈成纤维细胞-脱细胞真皮基质复合物(实验组)、犬牙龈成纤维细胞-脱细胞真皮基质复合物(对照组)及脱细胞真皮基质(空白组)分别植入裸鼠背部皮下,植入后2,4,8周,取背部标本进行组织学观察。结果与结论：人血小板源性生长因子B基因转染与未转染的犬牙龈成纤维细胞均能在脱细胞真皮基质上良好生长。植入后8周,空白组周围的细胞大面积进入脱细胞真皮基质内,部分脱细胞真皮基质出现完全自体化,尽管细胞进入较多,但新生成的胶原纤维较少,细胞只是占据原有的胶原支架生长;对照组开始出现大面积新生胶原纤维,脱细胞真皮基质上原有的胶原纤维逐步被新生的胶原替代,但原有的胶原结构得到保留;实验组出现大面积完全矿化,可见沿原有胶原支架排列的矿化颗粒。表明接种人血小板源性生长因子B基因修饰牙龈成纤维细胞的脱细胞真皮基质在体内获得了成骨性能。 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     

甲状腺脱细胞生物支架的制备及鉴定

目的 采用十二烷基硫酸钠（SDS）浸泡联合平板振荡制备兔甲状腺脱细胞生物支架,并对脱细胞支架进行评估。方法 20只新西兰大白兔随机分为正常对照组和脱细胞组,每组10只,脱细胞组兔甲状腺组织浸泡于1%SDS溶液后放置于37.0 ℃恒温振荡器中振荡24 h,去离子水振荡清除支架内残留去细胞试剂及DNA成分。对支架进行HE染色、DNA含量检测、扫描电子显微镜检查以及免疫荧光染色进行鉴定。 结果 甲状腺脱细胞支架DNA含量显著低于正常甲状腺组织（P<0.01）,DNA清除率达90%以上;经检测甲状腺脱细胞支架内无DNA成分残留,同时较为完整地保存了支架3D空间结构及蛋白成分。 结论 浸泡法联合平板振荡法可有效清除甲状腺细胞成分,较为完整地保留支架空间结构及蛋白成分。     

异体脱细胞真皮基质作为组织工程皮肤真皮支架的可行性*

背景：组织工程皮肤是目前研究皮肤损伤修复重建的重要手段之一,异体脱细胞真皮基质不存在免疫原性,在异体移植时不会发生排斥反应,是比较理想的真皮替代物。 目的：观察异体脱细胞真皮基质的组织相容性。 方法：以正常人体真皮组织作为对照,通过体外、体内细胞毒性实验检测异体脱细胞真皮基质的组织相容性,以膨胀度、饱和含水量及生物力学分析检测异体脱细胞真皮基质的亲水性及机械性能。 结果与结论：真皮基质中未见任何细胞成分,其网孔直径介于100~180 μm 之间。脱细胞真皮基质组饱和含水量为(69.6±3.97)%,膨胀度2.30±0.42,最大断裂力为(3.082±0.046) N,与对照组相比,差异无显著性意义(P > 0.05)。体内外细胞毒性检测,未见明显细胞生长抑制及免疫排斥反应。提示异体脱细胞真皮基质机械性能接近正常皮肤,组织相容性好,免疫排斥反应小,是构建组织工程皮肤理想的真皮材料。     

灌注法制备全肾脏脱细胞基质支架的体内生物相容性

背景：应用灌注法制备的大鼠全肾脏脱细胞基质支架具有良好的体外细胞相容性,但其体内生物相容性尚不明确。 目的：应用灌注法制备大鼠全肾脏脱细胞基质支架,检测其体内生物相容性。 方法：应用灌注法制备Wistar大鼠全肾脏脱细胞基质支架,进行以下实验：①急性毒性实验：在小鼠腹腔分别注射全肾脏脱细胞基质支架浸提液、生理盐水及苯酚。②溶血实验：将抗凝新西兰兔血分别与全肾脏脱细胞基质支架浸提液、生理盐水及蒸馏水混合。③热源实验：向新西兰兔耳缘静脉注射全肾脏脱细胞基质支架浸提液。④内皮刺激实验：在新西兰兔皮下注射全肾脏脱细胞基质支架浸提液,观察有无皮肤刺激反应。⑤皮下植入实验：将全肾脏脱细胞基质支架埋入新西兰兔背部皮下。 结果与结论：全灌注法制备的Wistar大鼠全肾脏脱细胞基质支架无细胞残留,未引起全身毒性反应、急性溶血反应、热源反应及皮肤刺激反应,植入兔体内具有良好的组织相容性。说明大鼠全肾脏脱细胞基质材料在动物体内具有很好的生物相容性。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：     

异体脱细胞真皮基质的研究进展

异体脱细胞真皮基质是天然真皮的脱细胞基质，经过冷冻干燥形成的三维支架结构。作为一种新兴的生物工程支架，它通过空间诱导和组织替代作用修复组织缺损。由于其源于人体正常组织，经临床应用证实，异体脱细胞真皮基质拥有优秀的组织相容性，现已广泛地应用于临床各科的创伤修复。     

NaOH消蚀法制备脱细胞真皮基质修补腹壁疝

背景：与其他疝修补材料相比,脱细胞真皮基质具有易血管化、抗感染、从而可用替代传统补片用于感染腹壁缺损重建等特点。 目的：观察NaOH消蚀法制备的脱细胞真皮基质作为修补材料应用于腹疝的应用价值。 方法：以全厚猪皮制成脱细胞真皮基质,45只SD雄性大鼠制备腹壁疝模型,随机数字表法分为腹壁疝组：直接缝合皮肤,Marlex网组和脱细胞真皮基质组：分别应用大小为3.5 cm×4.0 cm的Marlex网和脱细胞真皮基质缝合皮肤;观察修复后1周时有无腹壁疝发生,脱细胞真皮基质组修复后1,2,3,5,10周分别取材,其中每周各组取4只用于苏木精-伊红染色,光镜观察。修复后第5周Marlex网组和脱细胞真皮基质组各取6只用于抗张力试验。 结果与结论：术后1周脱细胞真皮基质组与Marlex网组腹壁疝发生率均显著低于腹壁疝组(P < 0.001)。脱细胞真皮基质的胶原纤维无明显变化,即有少量成纤维细胞植入,术后2周可见新生血管,术后5周脱细胞真皮基质内部血管密度基本稳定。将单独脱细胞真皮基质片与Marlex网行抗张力试验,Marlex网的抗张力显著高于脱细胞真皮基质(P < 0.001)。但植入体内5周后,脱细胞真皮基质筋膜组织的抗张力高于Marlex-筋膜组织(P < 0.05)。提示复合消蚀制备的脱细胞真皮基质可以作为良好的疝修补材料。     

不同时期局部移植人脐带间充质干细胞修复脊髓损伤的组织学观察

BACKGROUND:Spinal cord injury (SCI) is a disease causing a variety of motor and sensory dysfunctions, abnormal muscle tone and pathological reflex. Clinically, human umbilical cord mesenchymal stem cell transplantation has become an employed therapy for SCI. OBJECTIVE:To investigate the effect of local transplantation of human umbilical cord mesenchymal stem cells in different time after spinal cord injury in rats. METHODS:100 SPF male adult Sprague-Dawley rats were randomly divided into five groups: blank control group, SCI group, post-SCI 3-, 7-, 21-day transplantation groups (n=20 per group). Animal models of T₁₀ SCI were made by Allen’s method in the latter four groups, and rats in the three transplantation groups were given HUCMSCs transplantation at 3, 7, 21 days after SCI, respectively. RESULTS AND CONCLUSION:Compared with the SCI group, improved motor function scores, decreased interleukin-2 level, and increased serum interleukin-10 level were observed in the three transplantation groups at 49 days after modeling, indicating SCI was improved significantly in the three transplantation groups, especially in the post-SCI 7-day transplantation group. These findings suggest that human umbilical cord mesenchymal stem cell transplantation for SCI repair improves the movement function of rats, and cell transplantation at 7 days after modeling has achieved best outcomes.     

胰岛素样生长因子1诱导骨髓间充质干细胞转化为少突胶质细胞移植修复急性脊髓损伤

BACKGROUND:Treatment and rehabilitation of spinal cord injury is a complicated problem, and the reconstruction and remyelination of neural reflex pathways are the essential process, during which oligodendrocytes play an important role in spinal cord injury repair. OBJECTIVE:To observe the effect of oligodendrocyte transplantation for acute spinal cord injury in rats. METHODS:Insulin-like growth factor 1 induced bone marrow mesenchymal stem cells differentiating into oligodendrocytes, and those oligodendrocytes were transplanted into rats with acute spinal cord injury as induced cell transplantation group. Simple normal saline and natural oligodendrocytes were transplanted into the rat injured spinal cord as control group and oligodendrocyte group, respectively. Rat behavioral changes were observed by inclined plane test and Basso-Beattie-Bresnahan (BBB) scores. Neurological recovery and survival of the transplanted cells was detected and observed using spinal evoked potential and immunohistochemical staining, respectively. RESULTS AND CONCLUSION:Compared with the control group, BBB scores and the critical elevation angle of the incline plane test significantly increased, latencies of spinal motor and sensory evoked potential were on the decline (P < 0.05), and there were no significant differences in above indicators between the two groups at 4 and 8 weeks after transplantation. Moreover, survived oligodendrocytes after transplantation could be found in the lesions of spinal cord in both two groups. In conclusion, insulin-like growth factor 1-induced bone marrow mesenchymal stem cells can differentiate into oligodendrocytes that exact an excellent role in acute spinal cord injury repair after transplantation, which achieve the equal clinical efficacy to the natural oligodendrocytes.     

舒芬太尼预处理对脊髓损伤模型小鼠的神经保护

BACKGROUND: Sufentanil exerts protective effects on tissues, but its roles in the repair of nervous system injury and the underlying mechanism are still unknown.     

他莫昔芬对急性脊髓损伤模型大鼠的神经保护

BACKGROUND:Tamoxifen has been found to exert neuroprotection by reducing cerebral hemorrhage and edema surrounding the injured site of the spinal cord. OBJECTIVE:To investigate the neuroprotective effect of tamoxifen on rat acute spinal cord injury and the underlying mechanism. METHODS: Sixty Sprague-Dawley rats were equivalently randomized into five groups, and modeled into spinal cord injury at T10 level using modified Allen’s weight-drop method (70 g/cm), except those in sham operation group. At 30 minutes after modeling, all rats were given the intraperitoneal injection of 2.5, 5.0 and 10 mg/kg tamoxifen or same amount of normal saline, once daily. Basso, Beattie, Bresnahan (BBB) scores were recorded at 24, 48 and 72 hours after surgery. The injured spinal cord was removed at 72 hours to observe its edema. Meanwhile, the levels of interleukin-1β, interleukin-10 and tumor necrosis factor-α, as well as Caspase-3 activity were detected by ELISA; the protein levels of nuclear factor-κB p65, phosphorylated I-κBα and Caspase-3 were detected by western blot assay. RESULTS AND CONCLUSION: Compared with the model group, the hind limb function in the tamoxifen groups was significantly improved. Tamoxifen significantly decreased the water content in the rat spinal cord and inhibited spinal cord edema at 72 hours after surgery. ELISA results showed that tamoxifen significantly reduced the activity of interleukin-1β, interleukin-10, tumor necrosis factor-α and Caspase-3 (P < 0.05). Western blot assay revealed that tamoxifen significantly downregulated the expression levels of nuclear factor-κB p65, phosphorylated I-κBα and Caspase-3. These results suggest that tamoxifen protects against spinal cord injury via suppressing inflammatory response and apoptosis-associated proteins. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

多巴胺和5-羟色胺受体系统参与脊髓损伤后继发性骨质疏松症发病：新理论和诊疗策略

BACKGROUND:Current basic and clinical research have showed that increases in bone resorption and bone loss accur earlier after spinal cord injury (SCI) than disuse atrophy, revealing that other mechanisms are involved in the pathogenesis of the SCI-induced osteoporosis (SIO). OBJECTIVE:To introduce the current lab and clinical research progress in SIO focusing on the functional changes of two major neurotransmitters in the spinal cord, including dopamine (DA), 5-hydroxytryptamine (5-HT) and their receptors, as well as their regulatory functions on bone metabolism, aiming at finding a new treatment strategy for SIO. METHODS:A computer-based online search in PubMed and Embase databases was conducted for clinical and basic research related to SIO published from January 1967 to August 2016, using the keywords of “spinal cord injury; osteoporosis; dopamine; serotonin; 5-hydroxytryptamine” in English. Irrelevant, poorly related and repetitive studies were excluded, and finally 41 eligible articles were included. RESULTS AND CONCLUSION:DA and 5-HT are major neurotransmitters in the central nervous system, both involving in the regulation of bone remodeling. After SCI, loss of innervation and descending neurotransmitters especially DA, 5-HT and subsequent deregulation of their receptors are responsible for the onset of post-traumatic bone loss. The above research progress, in combination with the emerging clinical and lab investigations targeting 5-HT, DA and their receptors for improving neural functions after SCI, provides possible therapeutic pathways for SIO. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

甲基强的松龙影响脊髓损伤后内源性神经干细胞的迁移

BACKGROUND:After spinal cord injury, endogenous neural stem cells are activated to proliferate and migrate to repair damaged tissue. As a clinical medicine, methylprednisolone shows a lot of functions, but its effects on endogenous neural stem cells are still unknown. OBJECTIVE:To explore the effects of methylprednisolone on the proliferation and migration of endogenous neural stem cells after spinal cord injury. METHODS:Seventy-five Sprague-Dawley rats were used to make animal models of T10 complete paraplegia using Allen’s method, and randomized into methylprednisolone, normal saline and model groups. Rats in these three groups were given intraperitoneal injection of 1 g/L methylprednisolone solution at a dose of 30 mg/kg for 10 minutes and at a dose of 5.4 mg/kg/h for 23 hours, given intraperitoneal injection of normal saline at the same dose and given no treatment, respectively. Neurological and motor functions were assessed by somatosensory evoked potential and Basso Beattie Bresnahan scores at 7, 14, 21, 28 days after spinal cord injury. BrdU and Nestin staining of the injured spinal cord segment was conducted. RESULTS AND CONCLUSION:A large amount of BrdU- and Nestin-positive cells were visible in all the groups, and the number of these cells reached the peach at 14 days after spinal cord injury. Methylprednisolone was found to inhibit BrdU-, Nestin- or double-positive cells, indicating methylprednisolone can inhibit the proliferation and migration of endogenous neural stem cells. The results of Basso Beattie Bresnahan scores showed no notable improvement in the motor function of the limbs. Methylprednisolone also showed no significant effects on the motor evoked potential latency, but promoted nerve conduction recovery. All these findings indicate that methylprednisolone has some hindering effects on spinal cord repair by inhibiting the proliferation and migration of endogenous neural stem cells after spinal cord injury.     

脊髓康对脊髓损伤大鼠Nogo-66受体NgR表达的影响

文题释义：   文题释义：脊髓损伤：是指由于外界直接或间接因素导致脊髓损伤,在损害的相应节段出现各种运动、感觉和括约肌功能障碍,肌张力异常及病理反射等的相应改变。脊髓损伤的程度和临床表现取决于原发性损伤的部位和性质。脊髓损伤可分为原发性脊髓损伤与继发性脊髓损伤。前者是指外力直接或间接作用于脊髓所造成的损伤。后者是指外力所造成的脊髓水肿、椎管内小血管出血形成血肿、压缩性骨折以及破碎的椎间盘组织等形成脊髓压迫所造成的脊髓的进一步损害。Nogo受体NgR：是Nogo-A发挥作用的受体,是一种糖基磷脂酰肌醇锚定膜蛋白,能与细胞外结构域Nogo-66结合,并被激活,激活的NgR启动神经细胞内的信号转导通路抑制轴突再生和结构的重塑性。  背景：研究发现,脊髓损伤后髓鞘相关抑制分子的产生是影响轴突再生微环境的重要原因。中医药具有多因素、多靶点的优点,正逐步成为改善神经再生微环境的研究热点。目的：探索中药脊髓康对脊髓损伤后Nogo-66受体NgR表达的影响。方法：144只大鼠随机等分为6组,假手术组、模型组、强的松组、脊髓康高、中、低剂量组,每组24只。后5组以改良Allen’s法建立脊髓损伤动物模型。强的松组大鼠每天给予0.06 g/kg醋酸泼尼松灌胃,1次/d。脊髓康低中高剂量组大鼠每天给予12.5,25,50 g/kg脊髓康灌胃,1次/d。假手术组和模型组大鼠每天给予20 mL生理盐水灌胃,1次/d。各组动物均连续给药至动物处死。结果与结论：与模型组相比,强的松组及脊髓康低中剂量组大鼠脊髓组织中NgR蛋白及mRNA表达水平均显著降低。提示脊髓康可促进大鼠脊髓损伤后神经功能的恢复,可有效抑制脊髓损伤区NgR蛋白的表达,从而抑制不利于神经再生的因素,进一步改善神经再生的微环境。中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程 ORCID:0000-0002-8518-6160(郭杨)     

脊髓损伤模型大鼠胶质细胞反应性增生的变化规律及意义

BACKGROUND: The nervous reconstruction and repair after spinal cord injury have become a research hotspot.     

脊髓损伤后的康复实践结果

背景：脊髓损伤患者的康复结果与患者的损伤程度、治疗方法、康复时间及后期治疗等多因素有关。跨学科、全面、专业的脊髓损伤康复单元能为脊髓损伤患者提供更好的恢复。目的：综合评价脊髓损伤康复单元干预或其组合干预的效果。方法：检索2003至2014年Springer及PubMed数据库,检索词：spinal cord injury,rehabilitation practice,outcomes。根据纳入排除标准,阅读标题和摘要进行初筛,选用44篇英文文献进行分析。结果与结论：许多研究采用基于实践证据的方法,识别多种康复实践方法,把信息与结果联系以评价康复干预的效果。研究显示创伤及非创伤性混合样本中的目标实现与年龄没有差异,大多数康复结果很少有性别差异。最初入院到专科脊髓损伤中心的绝大多数脊髓损伤患者的并发症通常最低,患者尽早入住到跨学科、全面、专业的脊髓损伤单位比延迟入住缩短住院总时间。脊髓损伤患者接受正规、全面的门诊医护随访,在健康感知、独立性、抑郁症没有显著差异,但特定继发情况出现频率显著减少、程度减低。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

碱性成纤维细胞生长因子促进脊髓损伤小鼠内源性神经干细胞的增殖

BACKGROUND:So far steroid pulse therapy and surgical decompression are the main accepted therapies for spinal cord injury, but these methods make no effects on injured neurons. Nerve growth factors and endogenous neural stem cells play a role in the neuronal regeneration and remyelination, which provides a new idea for spinal cord injury treatment. OBJECTIVE:To explore the effect of the basic fibroblast growth factor on proliferation of endogenous neural stem cells after spinal cord injury and to analyze its relationship with the increased fluoro-gold labeled neurons. METHODS:Totally 48 Kunming mice were randomly divided into four groups including normal, spinal cord injury, treatment and sham operation groups. Acute spinal cord injury models were established in the spinal cord injury and treatment groups, and the normal group was subjected to operation that did not damage the spinal cord. At 2 hours after regaining consciousness, the treatment group was given daily injection of MTPBS containing 25 μg/kg basic fibroblast growth factors and 1% album. And at 7 days, laminectomy was carried out again at the L1 segment in the former three groups and a small piece of sterile gelfoam soaked with fluoro-gold was inserted into the incision. The sham operation group was given no processing. Afterwards, mouse motor behavior was assessed using Rotarod and Platform Hang tests; neurons in the corticospinal and rubrospinal tracts were labeled with fluoro-gold; the number of endogenous neural stem cells positive for nestin was detected by immunohistochemistry method. Besides, the correlation between the number of fluoro-gold labeled neurons and the number of endogenous neural stem cells was assessed. RESULTS AND CONCLUSION:The basic fibroblast growth factor could significantly improve the mouse motor behavior after spinal cord injury. And the number of endogenous neural stem cells was significantly increased after the basic fibroblast growth factor injection, which was related to the increased fluoro-gold labeled neurons. In conclusion, basic fibroblast growth factors play an important role in the proliferation of endogenous neural stem cells after spinal cord injury. Furthermore, endogenous neural stem cells improve locomotive behaviors by encouraging the neuronal proliferation.