神经病理性疼痛的干细胞治疗研究进展

<正>神经病理性疼痛(neuropathic pain,NP)是指原发性躯体感觉系统损害或疾病所致的疼痛[1]。周围神经或中枢神经系统的损伤均可诱发NP,如糖尿病周围神经病变、疱疹病毒感染、脊髓炎及脑卒中等。NP主要临床表现为自发性疼痛、诱发痛及痛觉过敏。当前NP的治疗主要包括药物治疗及非药物治疗两大类,常用的药物有三环类抗抑郁剂、5-羟色胺-     

移植GDNF基因修饰的神经干细胞对大鼠脑卒中的神经保护作用

目的:探讨移植胶质细胞源性神经营养因子(GDNF)基因修饰的神经干细胞(NSCs)是否比单纯NSCs移植对脑卒中有更好的神经保护.方法:用GDNF重组质粒转染NSCs,构建过表达GDNF的NSCs(GDNF/NSCs).插线法制备大鼠脑缺血卒中模型,再灌注3d,脑立体定位分别移植NSCs、 GDNF/NSCs以及生理盐水到同侧侧脑室,实验分为GDNF/NSCs组、NSCs组和对照组.再灌注第1、 2、 3、 5和7周处死大鼠,用改良的神经功能缺失评分和H-E染色分别评估大鼠神经功能和脑梗死体积;免疫组织化学显色观察移植的NSCs数量与分布和突触蛋白Syn、 PSD-95在脑组织的表达.结果:再灌注2、 3周,GDNF/NSCs组较NSCs组神经功能恢复更好.在各时间点GDNF/NSCs和NSCs组大鼠脑缺血体积较对照组显著减小;GDNF/NSCs组的NSCs细胞数量较NSCs组显著增多.再灌注2和3周,GDNF/NSCs组Syn免疫阳性产物比NSCs组显著增加;各时间点GDNF/NSCs组的PSD-95免疫阳性产物比NSCs组显著增加.结论:GDNF基因修饰的NSCs比NSCs对大鼠脑卒中有更好的神经保护作用.     

坐骨神经损伤大鼠模型鞘内移植神经干细胞后脊髓背角和背根神经节脑源性神经营养因子的表达

背景：坐骨神经损伤模型可测试伤害性的热刺激和机械刺激所引发的痛觉过敏及冷、触觉异常。 目的：观察坐骨神经损伤模型大鼠鞘内移植神经干细胞后脊髓背角和背根神经节脑源性神经营养因子的表达。 方法：72只SD大鼠随机均分为假手术组、对照组和实验组。对照组和实验组制作坐骨神经损伤模型,假手术组仅暴露坐骨神经,不结扎。分别于造模后第3,10天进行鞘内移植,实验组注入30 μL的神经干细胞悬液,空白组和对照组注入30 μL的细胞培养液。 结果与结论：与假手术组相比,对照组和实验组移植后3 d机械痛阈和热痛阈逐渐降低,至移植后7 d降低至最低点(P < 0.01),于移植后21 d恢复至移植前水平;实验组移植后7,14 d机械痛阈和热痛阈较对照组明显上升(P < 0.01)。与对照组相比,假手术组移植后7,14,21 d各组大鼠脑源性神经营养因子的表达呈低水平(P < 0.05);移植后14,21 d,实验组脑源性神经营养因子的表达量高于对照组(P < 0.05)。提示鞘内移植神经干细胞可提高脊髓背角和背根神经节中脑源性神经营养因子的表达。从而抑制了周围神经损伤产生的神经病理性疼痛。 关键词：脑源性神经营养因子;神经干细胞;慢性限制损伤;脊髓背角;背根神经节 doi:10.3969/j.issn.1673-8225.2012.10.031     

微囊化兔坐骨神经组织细胞移植对脊髓损伤后大鼠GDNF表达的影响

目的探讨微囊化兔坐骨神经组织细胞移植对脊髓半横断损伤后大鼠胶质细胞源性神经营养因子（GDNF）的表达影响。方法成年SD大鼠80只随机分为4组,微囊组（n=25）、细胞组（n=25）、单损组（n=25）、正常组（n=5）。术前制备兔坐骨神经细胞混悬液以及将其微囊化,微囊组、细胞组和单损组大鼠在脊髓半横断伤后,立即在损伤处分别植入10μl微囊化坐骨神经组织细胞1、0μl坐骨神经组织细胞以及10μl生理盐水。于术后2 d7、d1、4 d、21 d2、8 d（每个时相取5只大鼠）取出损伤部位脊髓组织,正常组（每个时相取1只大鼠）则取相应节段脊髓。组织石蜡包埋后切片,行免疫组织化学染色观察GDNF的表达变化。结果 GDNF阳性染色主要见于神经元细胞及胶质细胞的胞浆内。大鼠脊髓损伤后上述细胞2 d后表达开始上升2,1 d达到最高。微囊组与单损组、细胞组比较差异有显著性（P〈0.05）。结论微囊化兔坐骨神经组织细胞移植于大鼠损伤脊髓后,可以抑制炎症反应,促进GDNF的表达,有利于大鼠运动功能恢复。     

胶质细胞源性神经营养因子基因修饰骨髓基质干细胞移植脑出血大鼠 神经营养因子的表达

背景：研究证实,细胞移植和神经营养因子相结合治疗脑损伤能促进大鼠神经功能的恢复。 目的：观察移植胶质细胞源性神经营养因子基因修饰的骨髓基质干细胞对大鼠脑出血后神经营养因子表达的影响。 方法：通过脑立体定位仪向SD大鼠脑尾壳核注射胶原酶和肝素建立脑出血动物模型,将48只模型鼠随机分为3组,骨髓基质干细胞组、胶质细胞源性神经营养因子/骨髓基质干细胞组和对照组于建模后第3天在脑出血部位分别移植骨髓基质干细胞、胶质细胞源性神经营养因子/骨髓基质干细胞以及生理盐水。 结果与结论：与对照组和骨髓基质干细胞组相比,胶质细胞源性神经营养因子/骨髓基质干细胞组大鼠神经功能恢复更好;与对照组相比,移植后1,2周其他2组各神经营养因子表达均显著增加(P < 0.05)。提示胶质细胞源性神经营养因子基因修饰的骨髓基质干细胞移植治疗脑出血大鼠比单纯骨髓基质干细胞有更好的神经保护作用。     

移植BDNF和GDNF基因修饰的hMSCs对大鼠大脑中动脉阻塞的影响

 目的：构建脑源性神经营养因子（BDNF）和胶质细胞源性神经营养因子（GDNF）基因的非病毒表达载体,用脂质体法转染人骨髓间充质干细胞（hMSCs）,观察其对大鼠大脑中动脉阻塞（MCAO）模型的影响,探索移植转基因修饰的hMSCs治疗脑血管疾病的可行性。方法：构建高效非病毒表达载体,用脂质体法转染获得高表达2种神经营养因子的hMSCs。建立大鼠MCAO模型,建模后24 h经股静脉进行转基因hMSCs移植,并以磷酸盐缓冲液(PBS)和hMSCs为对照。用脑梗死体积计算、体重变化、行为学评测等指标对大鼠脑损伤程度进行评估,通过大鼠脑组织观察和病理切片对脑组织损伤以及细胞的迁移分化情况进行分析。结果：经股静脉转基因hMSCs移植能够提高大鼠MCAO后的感觉运动功能,减小脑梗死体积,与PBS对照组相比有显著差异;与hMSCs治疗组相比,治疗效果较好且稳定。移植的细胞在脑损伤区域有少数存活但未见分化现象。结论：经静脉移植脂质体介导、GDNF和BDNF基因修饰的hMSCs,可促进缺血脑组织的损伤修复,效果较好,为非病毒载体在干细胞相关转基因治疗的应用提供了理论依据。本研究表明,MSCs的作用不依赖干细胞的分化和神经元的替换,而可能与其分泌细胞因子对抗脑损伤并促进神经修复有关,在MSCs中转入特定的外源性神经营养因子可加强这一作用。     

神经干细胞移植修复大鼠脊髓半切伤的研究

目的：观察神经干细胞移植修复脊髓半切伤的疗效并探讨移植最佳时机。方法：制备大鼠T13-L1脊髓半切伤模型，取胎龄13、5d胎鼠大脑组织，体外分离、培养、诱导神经干细胞，并采用免疫细胞化学技术分别检测NSC（neural stemcell）特征性标志Nestin（巢蛋白）表达和用血清诱导分化为大量神经元NSE（neuron specific enolas）和神经胶质细胞GFAP（glial fibrillary acidic protein）表达。用明胶海绵吸附BrdU（5-溴脱氧尿嘧啶）标记好的神经干细胞悬液移植到脊髓半横断处，移植时间选择损伤后立即移植、损伤后第9、14d。观察移植后的大鼠行为的变化，通过CBS（combine behavioral score）评分和爬网格实验评价大鼠的运动功能恢复情况，并进行免疫组化鉴定，光学显微镜观察移植神经干细胞的存活和迁移。结果：在上述条件下培养及传代的细胞不断分裂增殖，形成悬浮生长的呈Nestin阳性的神经球；用血清诱导分化为大量表达NSE阳性的神经元和GFAP阳性的神经胶质细胞。与对照组相比，神经干细胞移植组明显修复了损伤结构，改善了下肢的功能，尤其是第9d移植组。结论：神经干细胞移植促进了脊髓损伤后神经结构和功能的恢复，是治疗脊髓半切伤一种有效方案，考虑综合因素移植干细胞的最佳时机应选损伤后9d左右。     

过表达胶质细胞神经营养因子基因转染骨髓间充质干细胞移植治疗脊髓损伤

文题释义： 胶质细胞源性神经营养因子：作为轴突再生的一种重要神经营养因子,可诱导间充质干细胞向神经样细胞分化并对中枢神经系统退行性疾病、脊髓损伤后神经功能恢复起到重要作用。 突触素：作为突触的特异性蛋白是突触形成过程中最重要的标志物,主要位于神经元胞体及轴突,可调节神经元轴突延伸,参与突触囊泡的介导转运、神经递质释放,对促进脊髓损伤后神经功能恢复起到重要作用。 背景：胶质细胞神经营养因子(glial cell line derived neurotrophic factor,GDNF)在诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)体外定向分化及促进脊髓损伤大鼠神经功能恢复过程中起到重要作用。 目的：观察过表达GDNF基因的BMSCs分化情况及其促进脊髓损伤大鼠神经功能恢复的潜在分子机制。 方法：①以重组目的基因腺病毒转染BMSCs并分为Ad-GDNF-GFP转染组、Ad-GFP转染组、未转染组,免疫荧光鉴定各组细胞神经元特异性烯醇化酶及微管相关蛋白2的表达,Western blot检测各组细胞GDNF、Wnt3a、Wnt7a蛋白表达。②以改良Allen法制备大鼠脊髓损伤模型,将造模成功的45只SD大鼠随机分为3组,分别以过表达GDNF基因BMSCs(GDNF-BMSCs)、BMSCs、PBS移植至脊髓损伤局部。移植后4周采用BBB评分法评估大鼠运动功能恢复情况,苏木精-伊红染色观察脊髓形态变化,免疫组化检测损伤局部神经元特异性烯醇化酶、突触素Ⅰ及胶质纤维酸性蛋白表达,Western blot检测损伤局部Bcl-2、肿瘤坏死因子α蛋白表达。 结果与结论：①Ad-GDNF-GFP转染组BMSCs可向神经元样细胞形态转变并表达神经元特异性烯醇化酶、微管相关蛋白2;Wnt3a、Wnt7a蛋白表达量显著高于Ad-GFP转染组、未转染组;②移植后4周,GDNF-BMSCs移植组大鼠BBB评分明显提高、脊髓空洞面积显著缩小。GDNF-BMSCs移植组脊髓损伤局部胶质纤维酸性蛋白、肿瘤坏死因子α表达量显著低于BMSCs移植组及PBS移植组,而神经元特异性烯醇化酶、突触素Ⅰ及Bcl-2表达量显著高于BMSCs移植组、PBS移植组;③结果表明,Wnt信号通路参与过表达GDNF基因 BMSCs向成熟神经元分化过程,移植后通过降低脊髓损伤局部炎症反应、减少细胞凋亡及胶质瘢痕形成、促进轴突再生,提高BMSCs移植治疗脊髓损伤的疗效。 ORCID: 0000-0001-6467-730X(黄成) 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     

脂肪源性干细胞促进大鼠受损坐骨神经传导及脊髓脑源性神经营养因子和睫状神经营养因子的表达

目的:探讨脂肪源性干细胞(ADSCs)对坐骨神经损伤大鼠神经传导功能以及脊髓脑源性神经营养因子(BDNF)和睫状神经营养因子(CNTF)表达的影响。方法:将第4代ADSCs移植入脱细胞神经移植物(ANA)中,构建组织工程神经。大鼠随机分为正常组、杜氏改良Eagle培养基营养混合物F12(DMEM)组和ADSC组。DMEM组和ADSC组均建立坐骨神经损伤模型,后用相应的组织工程神经桥接损伤神经的断端。术后6周采用神经电生理记录仪检测各组大鼠坐骨神经传导速度和波幅,采用免疫荧光和Real-time PCR检测各组大鼠脊髓脑源性神经营养因子(BDNF)、睫状神经营养因子(CNTF)蛋白和mRNA的表达。结果:ADSC组大鼠坐骨神经传导速度、波幅和脊髓BDNF和CNTF蛋白及mRNA表达均显著高于DMEM组。结论:ADSCs可增加坐骨神经传导速度和波幅、上调脊髓BDNF和CNTF的表达。     

神经干细胞联合胶质细胞源性神经营养因子脑内移植对Parkinson病大鼠纹状体内多巴胺含量的影响

将原代培养的胚鼠(E14.5)腹侧中脑神经干细胞(NSCs)单独或联合胶质细胞源性神经营养因子(GDNF)移植入Parkinson病(PD)大鼠纹状体内,术后各组动物存活一段时间,观察旋转行为后处死,取纹状体组织做高效液相色谱检测多巴胺(DA)含量。探讨NSCs单独或联合GDNF脑内移植对PD大鼠纹状体内DA含量的影响。结果显示:与PD模型相比,NSCs单独或联合GDNF脑内移植均能提高PD大鼠纹状体内的DA含量(P<0.05);在术后30d和60d,DA含量在GDNF+NSCs组的增加幅度比NSCs组更加明显,动物旋转行为亦得到明显改善(P<0.05)。上述结果表明,NSCs单独或联合GDNF移植均对PD大鼠有一定的治疗作用,而NSCs联合GDNF移植的治疗效果更好。     

经颈静脉移植神经干细胞治疗脑性瘫痪

BACKGROUND: Transplanted neural stem cells can survive, proliferate and differentiate into neurons and/or glial cells in the host, thereby promoting partial function recovery in the host.     

胚胎神经干细胞移植治疗阿尔茨海默病

BACKGROUND:More recently, stem cell therapy has become an issue of concern. Exogenous neural stem cell transplantation brings new hope for the treatment of nervous system injury by self-replication and differentiation to complement and replace damaged or dead nerve cells. OBJECTIVE:To explore the therapeutic efficacy of neural stem cell transplantation on Alzheimer’s disease. METHODS:Thirty APP/PS1 mice with Alzheimer’s disease were randomly assigned into model group, cell solution transplantation group or cell transplantation group (n=10 per group). Another 10 C57BL/6 mice were selected as controls. Embryos of C57BL/6 mice at 18 embryonic days were taken to make neural stem cell suspension followed by transfection using lentiviral vectors carrying GFP gene at different multiplicities of infection (1, 5, 10, 15, 20). Afterwards, GFP-transfected neural stem cells were implanted into the hippocampus of Alzheimer’s disease mice in the cell transplantation group, while the same volume of complete medium was injected into the hippocampus of mice in the cell solution transplantation group. Morris water maze test was performed at 2 weeks after cell transplantation, and brain tissues of mice was taken and detected histologically at 4 weeks after cell transplantation. RESULTS AND CONCLUSION:Compared with the control group, the escape latency was significantly higher, and the number of crossings over the target quadrant was lower in the other three groups (P < 0.05). Compared with the cell solution transplantation and model groups, in contrast, the escape latency was significantly lower, and the number of crossings over the target quadrant was significantly higher in the cell transplantation group (P < 0.05). Four weeks after transplantation, more intact neurons were found in the cell transplantation group as compared with the model group. These findings indicate that neural stem cell transplantation can improve behavior and morphology performance of mice with Alzheimer’s disease.     

人羊膜间充质干细胞移植修复肝脏缺血-再灌注损伤

BACKGROUND:Studies have shown that human amniotic mesenchymal stem cells can differentiate into hepatocyte-like cells, suggesting that human amniotic mesenchymal stem cell transplantation provides a new potential for the clinical treatment of liver diseases. OBJECTIVE:To observe the effect of human amniotic mesenchymal stem cell transplantation on the repair of liver ischemia-reperfusion injury repair. METHODS:Sixty Sprague-Dawley rats were randomized into stem cell transplantation, model and control groups. Animal models of liver ischemia-reperfusion injury were made in the rats in the stem cell transplantation and model groups. One hour after modeling, rats in the stem cell transplantation were given injection of human amniotic mesenchymal stem cells (0.5 mL, 106 cells) via the tail vein, while rats in the model and control group were given L-DMEM (0.5 mL) or normal saline (0.5 mL), respectively. Liver function and liver morphology were detected at 1, 2, 3 weeks after transplantation. Meanwhile, RT-PCR detection and western blot assay were also conducted. RESULTS AND CONCLUSION:(1) Liver function: Compared with the control group, levels of aspartate aminotransferase, alanine aminotransferase and malondialdehyde were significantly increased in the model group at different time points after transplantation (P < 0.05), while a significant reduction in the levels of these three indicators was found after cell transplantation as compared with the model group (P < 0.05). (2) Liver morphology: 2 weeks after transplantation, rats in the model group exhibited hepatocyte degeneration and necrosis, and severe fibrosis, but these changes were remarkably alleviated in the stem cell transplantation group. (3) PT-PCR and western blot detection: 2 weeks after transplantation, a significantly higher level of hepatocyte growth factor in the liver tissue and a lower level of α-smooth muscle protein were found in the stem cell transplantation group compared with the model group (P < 0.05). All these experimental findings indicate that human amniotic mesenchymal stem cell transplantation can improve impaired liver function in rats, possibly through regulating hepatocyte growth factor and α-smooth muscle protein expression levels in the liver, and thereby promotes the repair of liver ischemia-reperfusion injury.     

广谱抗菌药物防治造血干细胞移植后的早期感染

BACKGROUND:Occurrence of acute graft-versus-host disease after hematopoietic stem cell transplantation is closely related to early infection, so controlling infection can decrease the transplant- related mortality. OBJECTIVE:To explore effects of broad-spectrum antibiotics on the occurrence of early infection after hematopoietic stem cell transplantation. METHODS:Clinical data of 31 patients undergoing autologous peripheral blood stem cell transplantation were collected. Within 30 days after cell transplantation, occurrence rate and types of early infection were detected and recorded. Besides, distribution of pathogens, as well as treatment and outcome of patients were statistically observed. RESULTS AND CONCLUSION:All patients successfully underwent hematopoietic stem cell transplantation, and the occurrence rate of infection was 71% with no death at early stage after cell transplantation. Twenty strains of pathogens were detected, in which gram-negative bacteria accounted for 80%. In addition, there was a significant negative correlation between the number of neutrophils in the peripheral blood and the duration of infection (P < 0.01). These results indicate that the infection rate at early stage after hematopoietic stem cell transplantation is relatively higher, which is associated with reduction and recovery time of neutrophils. Therefore, it is advisable to choose appropriate broad-spectrum antibiotics for preventive treatment at early stage after cell transplantation, so as to quickly and effectively control infections.     

脐血间充质干细胞移植修复急性肠缺血再灌注损伤

BACKGROUND:Bone marrow mesenchymal stem cells can repair intestinal ischemia-reperfusion injuny by interfering inflammatory reactions after intestinal ischemia-reperfusion to protect intestinal barrier functions. In recent years, umbilical cord blood mesenchymal stem cells are gradually used as a substitute source of bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the effects of umbilical cord blood mesenchymal stem cells on acute intestinal ischemia-reperfusion injury. METHODS:Umbilical cord blood mesenchymal stem cells were induced, isolated in vitro and tracked by CM-DiI fluorescent labeling. Sixty-three Sprague-Dawley rats were equivalently randomized into three groups: control group received normal saline enema, intestinal ischemia-reperfusion injury group with ethanol diluted trinitro-benzene-sulfonic acid and transplantation group administrated with 1×10¹⁰/L umbilical cord blood mesenchymal stem cell suspension via the tail vein at 1 hour after trinitro-benzene-sulfonic acid modeling. At 3 days after transplantation, colon tissues were removed in each group to observe pathological changes of the intestinal tract by hematoxylin-eosin staining. Besides, expression of leptin mRNA in the colon tissues and cyclooxygenase-2 in the mucosa were detected by RT-PCR and immunohistochemistry method, respectively. RESULTS AND CONCLUSION:Transplanted umbilical cord blood mesenchymal stem cells distributed in the intestinal lymphoid tissue and among glandular epithelial cells, suggesting that these stem cells might be involved in the process of intestinal ischemia-reperfusion injury repair. Compared with the control group, intestinal injury in the injury group was significantly aggravated, and most intestinal epithelial cells shed; and the transplantation group appeared to have significantly reduced intestinal damage and significantly less cell shedding. Expression of leptin mRNA was significantly higher in the injury group than the transplantation group followed by the control group, and there were significant differences among the three groups (P < 0.05). Additionally, expression of cyclooxygenase-2 in the injury group was significantly higher than that in the control group (P < 0.05); compared with the injury group, expression of cyclooxygenase-2 was significantly lower in the transplantation group (P < 0.05). To conclude, leptin and cyclooxygenase-2 may be involved in acute intestinal ischemia-reperfusion injury, and umbilical cord blood mesenchymal stem cell transplantation significantly lessens intestinal ischemia-reperfusion injury, which provides an experimental basis for human treating acute intestinal ischemic injury.     

神经干细胞移植治疗老年痴呆

BACKGROUND:Neural stem cell transplantation has been used to treat a series of brain injury diseases, such as cerebral palsy, but its effect on Alzheimer’s disease is rarely reported. OBJECTIVE:To observe the effect of neural stem cell transplantation on the behavior and immune regulating system of Alzheimer’s disease rats.  METHODS:Thirty-five Sprague-Dawley rats were enrolled to make a postcerebral incision and given hippocampal injection of amanita phalloides acid to establish rat models of Alzheimer’s disease. Another 10 rats were only given hippocampal injection of normal saline after preparation of postcerebral skin incision as sham operation group. Then 32 successful rat models were randomly divided into two groups (n=16 per group): rats in experimental group were administrated hippocamal injection of 5×109/L allogeneic neural stem cell suspension; those in model group were given no injection. Five-day Morris water maze test was conducted at 4 weeks after transplantation. At 1 week after Morris water maze test, levels of interleukin-1 and interleukin-10 in the cerebral homogenate were detected, as well as pathological changes of brain tissues were observed in the three groups. RESULTS AND CONCLUSION:Compared with the model group, the abilities of cognition and memory were significantly higher in the sham operation group (P < 0.01), and the abilities of spatial learning and memory were significantly higher in the experimental group (P < 0.05, P < 0.01). Levels of interleukin-1 and interleukin-10 in the model group were significantly higher than those in the sham operation group (P < 0.01) but significantly lower than those in the experimental group (P < 0.01). Besides, the number of neurons in the model group was obviously less than that in the experimental and sham operation group. These results indicate that neural stem cell transplantation supplements and protects neurons against Alzheimer's disease in rats, thereby significantly improving the learning and memory ability.     

不同预处理策略促进间充质干细胞的归巢

BACKGROUND:Homing is the initial and key procedure of stem cells-based tissue restoration. Current studies have shown that the inability to recruit bone marrow mesenchymal stem cells to target tissue with high efficiency remains a significant barrier to tissue restoration. Preconditioning strategy provides a new insight to promote stem cell homing. OBJECTIVE:To review preconditioning strategies for promoting the homing of stem cells. METHODS:In PubMed database, different combinations of terms from “stem cell, mesenchymal stem cells, preconditioning, homing, migration” served as search terms to retrieve articles referring preconditioning strategies for promoting mesenchymal stem cell homing published from January 2000 to September 2015. According to the inclusion criteria, 72 articles were selected for final review. RESULTS AND CONCLUSION:Pretreating target tissue or mesenchymal stem cells ahead of cell transplantation, known as tissue preconditioning or cell preconditioning, prominently promotes the homing of mesenchymal stem cells, therefore enhancing tissue restoration effect. Tissue preconditioning is designed to up-regulate expression of chemokines by varying the local microenvironment, thereby increasing homing ability of mesechymal stem cells. Mesenchymal stem cell preconditioning strategies, for example, gene modification and cytokine induction, are mainly to up-regulate expression of chemokine receptors on the surface of mesenchymal stem cells as effectors, and thus promote targeted cell homing. Overall, preconditioning strategy will bring great hope to apply stem cell therapy into the clinic.     

脂肪源干细胞移植骨质疏松模型大鼠骨密度及骨形态计量学指标的变化

BACKGROUND:Stem cell transplantation is increasingly hoped to promote osteoblast differentiation and inhibit osteoclast proliferation in the treatment of osteoporosis. OBJECTIVE:To study the therapeutic effect of exogenous adipose-derived stem cell (ADSC) transplantation on osteoporosis in ovariectomized rats. METHODS:Thirty Sprague-Dawley female rats were equivalently randomized into sham, model, ADSC transplantation groups. Rats in all groups except the sham group underwent bilateral ovariectomy to make osteoporosis models. Surrounding adipose tissues instead of the ovary were removed in the sham group. After modeling, rats were given 2×10⁶ ADSCs at passage 4 via the tail vein in the transplantation group and the same volume of normal saline in the model group, once a week. After 6 weeks, levels of serum calcium, phosphorus, and alkaline phosphatase as well as bone mineral density and histomorphometry indicators were detected in rats. RESULTS AND CONCLUSION:Compared with the sham group, the trabecular bone volume fraction was significantly decreased in the model group (P < 0.01), but remarkably increased after ADSC transplantation (P < 0.05). After modeling, the bone trabecular absorption surface percentage and rate of bone trabecular formation were elevated significantly (P < 0.05 or P < 0.01), while these increases were improved by ADSC transplantation (P < 0.05). Additionally, the levels of serum calcium and alkaline phosphatase and bone mineral density were significantly decreased after modeling, but were increased after ADSC transplantation. In contrast, the serum level of phosphorus was significantly increased in the model group (P < 0.05) but decreased markedly in the ADSC transplantation group (P < 0.05). To conclude, ADSC transplantation can reduce the loss of bone mass in osteoporosis rats by ovariectomy.     

舒芬太尼复合丙泊酚全身麻醉在脐血间充质干细胞移植治疗脊髓损伤中的应用

BACKGROUND:Sufentanil and propofol are both found to have good neuroprotective effects on neurological damage in clinical practice. OBJECTIVE:To investigate the effect of propofol combined with sufentanil in umbilical cord blood mesenchymal stem cells transplantation for the treatment of spinal cord injury. METHODS:Sixty-five Wistar rats were selected to make animal models of acute spinal cord injury using Allen’s method. Six hours after modeling, these rats were randomly assigned into combined group (injection of 2×10⁷/L human umbilical cord blood mesenchymal stem cell suspension (0.5 mL) plus injection of 1.0-1.5 mg/kg propofol and 0.5 μg/kg sufentanil via the tail vein), stem cell group (injection of 2×10⁷/L human umbilical cord blood mesenchymal stem cell suspension (0.5 mL) via the tail vein), or control group (injection of 30 μL of LDMEM containing 5% fetal bovine serum). S100β protein level in serum was detected in each group at 15 and 60 minutes after injection. Motor function of rat in each group was assessed by Basso Beattie Bresnahan (BBB) scoring and incline plane test at 1, 2, 4, 6, 8 weeks after modeling. Pathological changes of the spinal cord were observed at 4 weeks after modeling. Expression of vascular endothelial growth factor was detected using western blot assay at 1 and 2 weeks after modeling. RESULTS AND CONCLUSION:After 15 and 60 minutes of intervention, S100β protein level was lowest in the combined group followed by the stem cell and control groups (P < 0.05). At 2, 4, 6, 8 weeks after modeling, scores on the incline plane test and BBB were ranked as follows: combined group > stem cell group > model group (P < 0.05). At 4 weeks after modeling, severe damage to the spinal cord and few nerve fibers were found in the control group; spinal cord hyperplasia and a few of regenerated axons and PKH-26-positive stem cells appeared in the stem cell group; while in the combined group, there were a large amount of PKH-26-positive stem cells and nerve axon-like structures. At 1 and 2 weeks after modeling, the highest protein level of vascular endothelial growth factor was found in the combined group followed by the stem cell group and control group (P < 0.05). To conclude, these findings indicate that propofol and sufentanil in umbilical cord blood mesenchymal stem cell transplantation therapy can promote the recovery of hindlimb function after spinal cord injury, thereby promoting the functional recovery of rats from spinal cord injury.     

视网膜干细胞移植对青光眼视网膜神经节细胞的保护

BACKGROUND:Stem cell transplantation is a new method for blinding eye disease. But there is a lack of research about the protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma. OBJECTIVE:To explore the protective effect of retinal stem cell transplantation on retinal ganglion cells of rats with glaucoma. METHODS:Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group) including control, model and retinal stem cell transplantation groups. Rat models of glaucoma were prepared in the latter two groups, and at 7 days after modeling, rats in the three groups were given intravitreal injection of 1 mL retinal stem cells (5x10⁶ cells), the same amount of PBS, and no treatment, respectively. Subsequently, relative indicators were detected at 2 weeks after transplantation. RESULTS AND CONCLUSION:The expressions of brain-derived neurotrophic factor and insulin-like growth factor I protein as well as the number of retinal ganglion cells were the highest in the control group, followed by the retinal stem cell transplantation group model group, and the lowest in the model group (P < 0.05). The number of apoptotic retinal ganglion cells in model group was significantly higher than that of control group (P < 0.05), and which in the retinal stem cell transplantation group was significantly lower than that in the model group (P < 0.05), but higher than that in the control group (P < 0.05). These results suggest that retinal stem cell transplantation for rat glaucoma can exert a protective effect on retinal ganglion cells.