骨髓间充质干细胞移植对肺气肿大鼠血浆TNF-α的作用

目的 研究移植骨髓间充质干细胞(MSCs)对肺气肿大鼠血浆肿瘤坏死因子-α(TNF-α)水平的影响.方法 将30只Wistar大鼠随机分为对照组、肺气肿组、肺气肿+MSCs治疗组,每组30只.在气管内注入脂多糖(LPS)并烟雾暴露,建立肺气肿大鼠模型,然后将培养的MSCs经尾静脉注入到肺气肿大鼠体内.观察移植后大鼠肺组织的变化,酶联免疫吸附测定(ELISA)法检测血浆TNF-α含量.结果 肺气肿组、肺气肿+MSCs移植组均出现肺气肿样病理改变,但后者较前者明显减轻.与肺气肿组比较,肺气肿+MSCs组TNF-α含量显著降低.结论 MSCs移植对肺气肿大鼠有保护作用.Abstract：Objective To investigate the effect of bone marrow mesenchymal stem cell (MSCs) transplantation on plasma TNF-α levels in rats of pulmonary emphysema. Methods Thirty wistar rats were randomized into three groupes:normal control group, pulmonary emphysema group, pulmonary emphysema +MSCs transplantation groupe. The pulmonary emphysema model of rats were established by intratracheal instillation of lipopolysaccharide twice and daily exposure to smog. Bone marrow MSCs were infused through tail vein. Morphologic changes of the lung tissues were observed. The level of plasma TNF-α were dectected using enzyme-linked immunosorbent assay. Results Destruction of alveolar walls was observed in rat lungs from pulmonary emphysema group and pulmonary emphysema +MSCs transplantation group. MSCs transplantation group significantly ameliorated the emphysematous changes. Significant differences in the levels of TNF-α between pulmonary emphysema group and pulmonary emphysema +MSCs transplantation group were observed. Conclusions MSCs therapy shows protective effects against pulmonary emphysema.     

自体骨髓间充质干细胞移植治疗心肌梗死后心力衰竭的实验及临床研究

目的 观察自体骨髓间充质干细胞(MSCs)移植对心肌梗死后心力衰竭的疗效.方法 选择21只中国小型家猪,应用闭胸经股动脉介入法制作心肌梗死后心力衰竭模型,术后动物左室射血分数(LVEF)<45%、B型钠脲肽(BNP)水平>100pg·ml-1为心力衰竭建模成功.随机将动物分为实验组(n=11)和对照组(n=10),术后2周内实验组及对照组经冠状动脉途径分别移植MSCs及磷酸盐缓冲液,8周后,行心脏彩超、单光子发射型计算机断层扫描(ECT)及BNP检查,观察心功能的改变.同时,选择心肌梗死致心力衰竭[左室射血分数(LVEF)<45%,BNP>100pg·ml-1]患者8例,发病2周内经皮冠状动脉途径植入MSCs,术前及术后8周行心脏彩超、ECT及BNP检查,监测心功能的变化.结果 动物实验组和对照组分别有8头和7头猪造模成功;心脏彩超结果显示:实验组MSCs移植后LVEF由(39±1.70)% 增加至 (51±3.52)%,ECT心血池显像显示LVEF由(35.08±3.12)% 升高至 (52.15±1.21)%,BNP水平由移植前的(298±17)pg·ml-1下降至(87±15)pg·ml-1;对照组以上指标无明显改变.临床观察显示:MSCs移植后患者心脏彩超显示LVEF由(34.06±2.13)%增加至(52.08±1.02)%,ECT心血池显像显示LVEF由(34.12±3.09)%增加至(53.03±1.01)%,BNP水平由(326±34)pg·ml-1降低至(101±7)pg·ml-1.结论 自体MSCs移植是治疗急性心肌梗死后心力衰竭安全、有效的新方法.     

人骨髓间质干细胞治疗实验性变态反应性脑脊髓炎大鼠的实验研究

目的:研究尾静脉注射入骨髓间质干细胞(hMSCs)移植治疗实验性变态反应性脑脊髓炎(EAE)大鼠的疗效及移植后hMSCs在EAE大鼠体内的状况.方法:分离和纯化hMSCs.用豚鼠脑脊髓匀浆免疫Wistar大鼠制备EAE大鼠模型,分别在免疫后10、15、21天经尾静脉注射移植未分化的hMSCs入大鼠体内,通过电镜,动物模型神经功能评分,病理观察hMSCs对EAE大鼠的疗效,利用免疫组化方法观察移植后的hMSCs在EAE大鼠体内存活、迁徙、分化的状况.结果:免疫后10天和15天hMSCs移植组大鼠自移植后10天起其神经功能评分显著低于对照组(P＜0.05);在电镜下治疗组髓鞘超微结构改变明显轻于对照组;病理上各移植组在移植后10、30天脑内脱髓鞘病灶数目明显少于对照组(P＜0.05).hMSCs移植组各时间点大鼠脑组织切片中均可见Brdu、NSE、GFAP、CNPase染色阳性细胞,CNPase( )的少突胶质细胞比例随时间的延长而逐渐增多.结论:hMSCs移植能有效改善EAE动物的神经功能评分,减少脱髓鞘病灶数目,并且在体内微环境改变信号的影响下分化成神经元、星型胶质细胞、少突胶质细胞.     

髓芯减压联合自体浓缩骨髓有核细胞移植治疗早期股骨头坏死

目的：观察髓芯减压联合自体浓缩骨髓有核细胞移植治疗早期股骨头坏死（ONFH）的临床疗效。方法回顾性分析该院收治的早期ONFH患者56例（82髋）,将患者分为观察组（30例,44髋）和对照组（26例,38髋）,观察组采用骨髓间充质干细胞移植联合髓芯减压治疗,对照组采用单纯髓芯减压治疗。通过比较两组患者术前和术后3、6、18个月髋关节Harris髋关节评分及术后18个月影像学变化、坏死面积百分比改变程度等,了解两种治疗方法的临床疗效。结果观察组Harris髋关节评分升高至（88．24±5．53）分,优良率达84．09％;对照组升高至（75．84±4．20）分,优良率为71．05％;MRI坏死面积：观察组逐渐减少到（13．86±4．27）％,对照组降至（19．53±5．29）％,两组比较差异有统计学意义（P＜0．05）。术后两组Harris髋关节评分和影像学变化均较术前有所改善（P＜0．05）。结论髓芯减压联合自体浓缩骨髓有核细胞移植治疗早期ONFH的临床疗效明显优于单纯髓芯减压。     

Lithium chloride prevents glucocorticoid-induced osteonecrosis of femoral heads and strengthens mesenchymal stem cell activity in rats

Background:Accumulating evidence suggests that lithium influences mesenchymal stem cell (MSC) proliferation and osteogenic differentiation. As decreased bone formation in femoral heads is induced by glucocorticoids (GCs), we hypothesized that lithium has a protective effect on GC-induced osteonecrosis of femoral heads (ONFH).Methods:A rat ONFH model was induced by methylprednisolone (MP) and the effect of lithium chloride on the models was evaluated. Micro-computed tomography (CT)-based angiography and bone scanning were performed to analyze the vessels and bone structure in the femoral heads. Hematoxylin and eosin and immunohistochemical staining were performed to evaluate the trabecular structure and osteocalcin (OCN) expression, respectively. Bone marrow-derived MSCs were isolated from the models, and their proliferative and osteogenic ability was evaluated. Western blotting and quantitative real-time polymerase chain reaction were performed to detect osteogenic-related proteins including Runx2, alkaline phosphatase, and Collagen I.Results:Micro-CT analysis showed a high degree of osteonecrotic changes in the rats that received only MP injection. Treatment with lithium reduced this significantly in rats that received lithium (MP + Li group); while 18/20 of the femoral heads in the MP showed severe osteonecrosis, only 5/20 in the MP + Li showed mild osteonecrotic changes. The MP + Li group also displayed a higher vessel volume than the MP group (0.2193 mm³vs. 0.0811 mm³, P < 0.05), shown by micro-CT-based angiography. Furthermore, histological analysis showed better trabecular structures and more OCN expression in the femoral heads of the MP + Li group compared with the MP group. The ex vivo investigation indicated higher proliferative and osteogenic ability and upregulated osteogenic-related proteins in MSCs extracted from rats in the MP + Li group than that in the MP group.Conclusions:We concluded that lithium chloride has a significant protective effect on GC-induced ONFH in rats and that lithium also enhances MSC proliferation and osteogenic differentiation in rats after GC administration.     

骨髓间充质干细胞治疗早期股骨头坏死的实验研究

目的:探讨骨髓间充质干细胞(MSC)治疗早期股骨头坏死的疗效及机制. 方法:60只新西兰大白兔造模后选取其中确诊为股骨头坏死的36只,随机分为3组:对照组:不予以任何处理;单纯减压组和减压+MSC移植组.术后4周及8周各组随机挑选6只兔先行MRI扫描,之后处死动物取股骨头行常规H-E染色及CD34免疫组化染色,用扫描电镜观察超微结构. 结果:术后第8周,减压+MSC移植组MRI示低信号区明显缩小,组织病理学及扫描电镜表现空骨陷窝减少,成骨细胞多,新骨形成,空骨陷窝阳性数和微血管密度与单纯减压组和对照组比较差异有统计学意义(P＜0.05). 结论:MSC移植能促进兔股骨头坏死的修复.     

Bcl-2基因转染骨髓干细胞治疗激素性股骨头坏死的疗效研究

研究Bcl-2 基因转染的骨髓间充质干细胞（BMSCs）结合髓芯减压法对兔早期激素性股骨头坏死的疗效。方法使用共计40只家兔复制早期激素性股骨头坏死模型,其中成功的模型家兔共28 只。随机分为4 组：空白对照组6 只,不作任何处理;髓芯减压组6 只,单纯减压;BMSCs 治疗组8 只,在减压同时植入BMSCs;Bcl-2 治疗组8 只,在减压同时植入Bcl-2 基因转染的BMSCs。植入后分别在第8、12 周各取股骨头标本行病理学检查及骨细胞凋亡检测,观察骨细胞生长及凋亡情况。结果成功复制兔早期激素性股骨头坏死动物模型;培养出BMSCs后进行传代;Bcl -2 基因转染成功;治疗12 周后,Bcl-2 治疗组动物的一般情况改善,组织切片和细胞凋亡检测结果显示,Bcl-2 治疗组的空骨陷窝和骨细胞凋亡数量与其他3 组比较,差异有统计学意义（p <0.05）。结论使用髓芯减压将Bcl-2 基因修饰的BMSCs 移植到兔早期激素性股骨头坏死动物模型体内,具有较好的成骨治疗作用。     

髓芯减压植骨+BMP治疗早期股骨头坏死

目的 探讨更有效的治疗早期股骨头坏死的方法.方法 对2002年1月～2004年4月在潍坊医学院附属医院应用髓芯减压植骨＋BMP治疗的15例（17个髋）股骨头坏死的患者进行回顾性的分析.男6例,女9例;年龄36～62岁,平均47岁,随访时间26～94个月,平均53个月.按改良的Ficat分期标准,ⅡA期15髋,ⅡB期1髋,Ⅲ期1髋.结果 采用改良Jacobs百分评价法作为疗效标准,优14个髋（12个病人,均为ⅡA期）,可2个髋,差1个髋.结论 髓芯减压植骨＋BMP能有效地治疗早期股骨头坏死.     

骨髓间充质干细胞在治疗股骨头坏死方面的临床研究

目的：探讨骨髓间充质干细胞移植技术在治疗股骨头坏死的疗效。方法选用齐齐哈尔医学院附属第三医院骨科股骨头坏死患者60例,按ARCO分期均为1～2期,其中采用死骨刮除＋植骨＋骨髓间质干细胞移植技术治疗股骨头坏死患者30例,采用传统死骨刮除＋植骨治疗股骨头坏死患者30例,通过放射线检查术后骨质变化情况和VAS疼痛评分及Harris 髋关节功能评分进行对比,对两种治疗方法疗效优良率采用χ2检验。结果两组患者住院时间及治疗效果比较差异显著,有统计学意义。结论骨髓间充质干细胞移植治疗股骨头坏死,能够有效恢复髋关节功能,减轻髋关节疼痛症状,改善股骨头坏死区域的血运,防止股骨头塌陷,保护股骨头,延缓髋关节置换的手术时间。     

VEGF-165基因修饰的骨髓间充质干细胞移植修复犬股骨头坏死

目的研究hVEGF-165基因转染的犬骨髓间充质干细胞移植修复自体股骨头坏死的效果。方法用股骨颈截骨后克氏针再固定的方法建立犬股骨头坏死的动物模型,分为转基因间充质干细胞移植组、单纯间充质干细胞移植组和未治疗组,每组9只动物(18侧髋关节)。脂质体法将hVEGF-165基因转染进入骨髓间充质干细胞。造模后10周经大转子将细胞植入股骨头坏死区,转基因间充质干细胞和未转基因干细胞随机植入同一动物的左侧或右侧后肢。细胞移植后第2、8和12周取股骨头标本进行骨小梁定量分析,结合X线和同位素断层扫描分析3组间成骨修复的差别,并比较第12周时各组标本中截骨平面以上的血管数。结果①造模手术后10周,手术侧股骨颈骨折已愈合,但股骨头均出现典型的坏死。②骨小梁定量分析提示,在每一个时间点各组的成骨修复的差别有统计学意义:转基因间充质干细胞移植组的成骨修复好于单纯干细胞移植,未治疗组最差。X线和SPECT的表现与此相符。③12周时,转基因间充质干细胞移植组中血管数量最多,间充质干细胞移植组与未治疗组的血管数量没有统计差别。结论与未转基因的间充质干细胞移植相比,转染hVEGF-165基因后的间充质干细胞移植能够增加股骨头坏死区血管...     

A modified technique of bone grafting pedicled with femoral quadratus for alcohol-induced osteonecrosis of the femoral head

Background Quadratus femoris pedicled bone grafting has yielded satisfactory long-term clinical outcome for osteonecrosis of the femoral head （ONFH） in pre-collapse ONFH without extensive lesion. However, for pre-collapse ONFH with extensive necrotic area, it is still challenging to preserve the femoral head. The current study aimed to introduce a new technique of deliquesce strut with titanium mesh containing bone grafting pedicled with the femoral quadratus and to evaluate its short-term outcomes.Methods From January 2008 to December 2008, 10 ONFH patients （12 hips） underwent operations by a new technique of deliquesce strut with titanium mesh containing bone grafting pedicled with the femoral quadratus （group A）.According to the ARCO classification system, there were two hips in stage Ⅱ B and 10 hips in stage Ⅱ C. Also in the same period, 12 ONFH patients （16 hips） underwent operations by the conventional procedure of quadratus femoris pedicled bone grafting （group B）. There were 6 hips in stage Ⅱ B and 10 hips in stage Ⅱ C. All patients were males and suffered from alcohol induced ONFH. For the new technique, the necrotic area was evaluated, and a titanium mesh piece of the same size （range from 2.5 cm×2.8 cm to 2.8 cm×3.4 cm） was obtained and shaped to match the contour of the head. The cancellous bone was first placed underneath the subchondral bone and was densely impacted （about 1 to 2 mm thick）.Then the titanium mesh piece was inserted. The length of the decompressive trough was measured. A titanium cylinder mesh cage with a diameter of 1.6 cm of the same length was obtained, with a ＂U＂ shaped window in the wall being created to make room for the muscle pedicle. The muscle pedicle bone was inserted into the titanium mesh cage to form a bone graft-titanium cage complex and, then the complex was inserted. The hundred percent score method was used for outcome evaluation. Clinical and radiographic outcomes were compared between group A and group B.Results The average operative time was 150 minutes （130 to 185 minutes） in group A, with an average of 130 minutes （120 to 180 minutes） in group B. The mean blood loss was 400 ml （300 to 500 ml） in group Aand 350 ml （250 to 500 ml）in group B. Group A patients were followed up for an average of 19.2 months （14 to 24 months）, with an average of 18.5 months （12 to 24 months） for Group B. Full weight bearing was allowed 5 to 7 months postoperatively. Pain and function were obviously improved. For group A, pain score improved from a mean of 9.8 points preoperatively to an average of 24.6 points postoperatively, and function score improved from a mean of 9.0 points preoperatively to an average of 17.4 points postoperatively. In group B, pain score improved from a mean of 9.5 points preoperatively to an average of 24.2 points postoperatively and function score improved from a mean of 9.2 points preoperatively to an average of 17.2 points postoperatively. The range of motion changed the least, with score improvement from a preoperative mean of 13.9 points to postoperative 16.8 points for group A and from a preoperative mean of 13.7 points to postoperative 16.5 points for group B. Radiographic score improved from preoperative 31 points to postoperative 38 points for group A, in comparison with an improvement from preoperative 31 points to postoperative 37 points for group B. At the latest follow up, 11 hips were rated as excellent and 1 hip was better for group A, with 14 hips being rated as excellent and 2 hips being better in group B. There was no statistically significant difference between groups A and B in clinical and radiographic outcomes.Conclusion For ONFH in stage ARCO llC, satisfactory clinical outcome can be achieved by the new technique in the short-term period while the long-term clinical outcome has yet to be determined.     

髓心减压并自体骨髓干细胞移植治疗创伤性股骨头坏死

目的观察髓心减压并自体骨髓干细胞移植治疗股骨头坏死的临床效果。方法9例股骨头坏死患者经骨髓干细胞动员后,第2天进行骨髓干细胞采集,经髓心减压后,将干细胞悬液缓慢匀速注入。干细胞移植后随访观察患者髋关节疼痛程度、性质及持续时间变化,5个月后行股骨头供血动脉造影术观察股骨头血供情况。结果9例患者随访10个月,髋关节疼痛有不同程度缓解,关节功能改善。干细胞移植5个月后,9例患者股骨头供血动脉数字减影血管造影严查,均显示旋股内动脉及闭孔动脉管径增粗,新生血管增多,血流速度增快,股骨头血液供应明显改善。治疗中均为发生严重的并发症和不良反应。结论干细胞移植治疗股骨头坏死方法简单,安全有效,适合Ⅰ-Ⅲ期早、中期股骨头坏死患者。     

BMSCs/EPCs联合移植治疗兔股骨头坏死的实验研究

目的 检测骨髓来源内皮祖细胞（endothelial progenitor cells,EPCs）与骨髓基质细胞（bone marrow mesenchymal stem cells,BMSCs）移植到股骨头缺血坏死部位促进坏死区局部成骨和成血管能力.方法 使用密度梯度离心法分离培养兔BMSCs、EPCs并鉴定.共培养细胞进行碱性磷酸酶活性检测以确定EPCs与BMSCs最佳比例.培养的细胞分成3组植入兔股骨头坏死模型：A组（单独移植BMSCs）、B组（单独移植EPCs）、C组（最佳比例联合移植BMSCs与EPCs）,于术后4周对兔股骨头坏死模型进行成骨及成血管指标检测.结果 EPCs与BMSCs的最佳共培养比例为1∶1.与A组及B组相比,C组有着更高的BMP-2蛋白表达和更多的新生血管形成（P＜0.05）.组织学检测显示：A组可见类骨质结构;B组可见血管结构,但类骨质生成较少;C组可见类骨质结构,并且可见密集的血管结构.结论 以1∶1比例联合移植EPCs和BMSCs到股骨头坏死部位具有很强的成骨以及成血管能力.