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1.
自体骨髓干细胞移植在肢体动脉缺血时的作用   总被引:13,自引:12,他引:1  
金毕  赵玉国  程丽  杜玉清  田元 《中华实验外科杂志》2004,21(11):1339-1340,i002
目的 探索一种简便安全有效的治疗下肢动脉缺血性疾病的方法。方法 建立鼠后肢缺血动物模型 ,将取于自体的骨髓干细胞 (BMSC)制成悬液注射于缺血部位肌肉内共 7点 ,每点间隔 0 .2cm ,4周后行动脉造影 ,并取肌肉标本测定毛细血管密度。结果 动脉造影显示治疗组缺血肢体侧枝动脉明显增多 ;毛细血管密度增加 ,每高倍镜视野达到 5个 ,与缺血组 (2个 /高倍镜 )相比 ,差异有显著性 (P <0 .0 5 )。结论 自体骨髓干细胞移植是一种新的、简单有效的治疗下肢缺血的方法。  相似文献   

2.
自体骨髓单核细胞移植对兔肝内缺血型胆道病变的影响   总被引:1,自引:1,他引:0  
目的 探讨自体骨髓单核细胞移植对血管新生及胆道缺血病变的影响.方法 将30只日本大耳白兔随机分为3组:假手术组(A组)、实验模型组(B组)和骨髓单核细胞移植组(C组),每组10只.3组均游离胆总管、肝总动脉及其间的疏松结缔组织,B组和C组阻断胆总管远端及肝总动脉2 h,C组经肝总动脉注射PKH26标记的自体骨髓单核细胞.术后监测生化指标的变化情况.术后4周开腹行胆道造影.同时,取第一肝门处汇管区肝组织制作石蜡切片,应用免疫组织化学方法观察自体骨髓单核细胞在肝内缺血环境中的分布及分化情况,并检测微血管密度.结果 术后C组生化指标恢复明显优于B组,骨髓单核细胞可分化成血管内皮细胞.术后4周,C组胆道破坏较B组轻,且胆道周围新生毛细血管多于B组.结论 自体骨髓单核细胞移植可以促进局部缺血组织毛细血管的增生,改善局部缺血胆道组织的微循环,从而减轻缺血型胆道病变的程度,甚至预防缺血型胆道病变的发生.  相似文献   

3.
pSV-VEGF165肌肉原位注射促进兔缺血后肢毛细血管形成   总被引:7,自引:0,他引:7  
Zhao Y  Shi Y  He T  Zhang H  Chen S  Zhang B 《中华外科杂志》1999,37(4):211-213
目的 探索一种更为简便安全的下肢动脉缺血性疾病的基因治疗途径。方法 建立兔后肢血动物模型,将体外构建的重组大肠杆菌质粒pSV-VEGF1651mg分5点注射于缺血肌群,30天后切取标本,测定毛细血管密度和毛细血管数/肌肉数比值。结果经pSV-VEGF166治疗后,缺血肢体毛细 管密度和毛细血管数/肌肉数比值显著增加,小腿部位比大腿部位增加明显(P〈0.05)。结论 肌肉内原位注射pSV-ESGF1  相似文献   

4.
目的观察血管内超声消融联合自体骨髓干细胞移植治疗下肢动脉硬化性闭塞症的疗效。方法分析39例下肢动脉硬化性闭塞症的患者行血管内超声消融联合自体骨髓干细胞移植治疗的临床资料。男21例,女18例;平均年龄63岁。手术首先抽取自体骨髓260mL,分离出单个核细胞,制备干细胞悬浊液40mL。采用经皮股动脉穿刺或切开显露,经股浅动脉上部向下插入超声消融导管,在动脉造影监视下进行血管内超声消融,尔后将干细胞悬浊液一次性分点注射于小腿缺血肌肉内局部。结果39例患者术后静息痛缓解,跛行距离增加,冷、凉感觉得到改善。结论血管内超声消融联合自体骨髓干细胞移植治疗下肢动脉硬化性闭塞症是一种简便、有效的方法。  相似文献   

5.
目的 探讨自体骨髓干细胞移植术治疗下肢动脉缺血性疾病的临床效果.方法 回顾性分析9例行下肢血管造影检查确诊的下肢动脉缺血患者(9条患肢)采用自体骨髓干细胞移植术治疗的临床资料.结果 经治疗后,有6例患者(6条患肢)静息痛缓解或消失,溃疡愈合,间歇性跛行消失或距离延长,3例患者效果不佳.结论 自体骨髓干细胞移植术治疗下肢缺血性疾病具有一定的疗效,方法简单、安全.  相似文献   

6.
目的观察血管内超声消融联合自体骨髓干细胞移植治疗糖尿病足的疗效。方法37例糖尿病足患者,手术首先抽取自体骨髓干细胞260ml,分离出单个核细胞,制备干细胞悬浊液40ml。采用经皮股动脉穿刺或切开显露,经股浅动脉上部向下插入超声消融导管,在血管造影监视下进行血管内超声消融后,将制备的干细胞悬浊液一次性分点注射于缺血的小腿肌肉内。结果37例患者术后间歇性跛行距离增加大于200m,趾端溃疡面积减小,冷、凉感觉得到改善。结论血管内超声消融联合自体骨髓干细胞移植治疗糖尿病足是一种简单、有效的方法。  相似文献   

7.
目的:探讨犬自体骨髓干细胞在缺血组织的新生血管形成中的作用.方法:抽取犬骨髓,经免疫磁珠系统分离出CD 34细胞,体外扩增并向血管内皮细胞诱导分化;建立犬双下肢缺血动物模型,将诱导分化细胞移植入一侧缺血肢体中作为实验组,另一侧缺血肢体植入等体积的生理盐水作为自体对照组.移植后6周,检测缺血肢体中新生血管的形成.结果:细胞移植后6周,动脉造影显示实验组缺血肢体侧枝循环增加,明显多于对照组.微血管密度检测实验组为(14±2.3)个/高倍镜视野,明显高于对照组(6±2.1)个/高倍镜视野(P<0.05).激光共聚焦显微镜证实缺血肢体中移植的自体骨髓干细胞参与了新生血管形成.结论:自体骨髓干细胞移植可以促进缺血肢体组织中的新生血管形成.  相似文献   

8.
自体骨髓干细胞移植治疗下肢动脉缺血性疾病的疗效观察   总被引:3,自引:2,他引:1  
目的:观察自体骨髓干细胞移植治疗下肢动脉缺血性疾病的临床疗效及影响因素。方法:28例下肢动脉缺血患者,均抽取自体骨髓血,体外提取单个核细胞,行缺血下肢局部肌肉注射,术后观察各项指标综合评估。将患者分为高浓度注射组和低浓度注射组,比较两组移植前后缺血改善情况。结果:28例患者总的疼痛缓解率及麻木感和冷感缓解率均为100%,5例溃疡出现愈合迹象,移植后临床评分(10.96±2.09)低于移植前(14.25±2.41),P<0.05;高浓度注射组注射前后评分差值(3.92±1.16)大于低浓度注射组(2.81±1.52,P<0.05。除1例于术后3个月出现浅静脉曲张外,未发现明显并发症。结论:自体骨髓干细胞移植治疗下肢动脉缺血性疾病是一种安全、有效的手段。干细胞提取的数量、动脉缺血的范围,尤其是干细胞注射浓度对于其治疗效果有重要影响。  相似文献   

9.
目的探讨粒细胞集落刺激因子(rhG-CSF)对下肢缺血模型血管新生的影响。方法制作兔左下肢缺血模型,术后随机分为rhG-CSF治疗实验组(n=24)和对照组(n=24);应用流式细胞学技术、动脉造影、免疫组织化学染色检查,比较两组外周血CD34 细胞的含量、缺血下肢侧枝血管计数及肌肉毛细血管密度。结果治疗后3 d实验组CD34 含量(%)为(0.7150±0.0873)明显高于对照组(0.3983±0.0853),差异有统计学意义(P<0.01);实验组在第15、30天时侧枝血管计数(6.33±0.82、9.17±0.75)均高于对照组(3.33±0.52、4.17±0.75)(P<0.01);第40天实验组内收肌毛细血管密度平均为8.5/HP,明显高于对照组4.2/HP(P<0.01)。结论rhG-CSF可以增加兔缺血下肢的毛细血管数量,有促进血管新生的作用。  相似文献   

10.
目的将基质细胞衍生因子(SDF-1)用于自体骨髓单个核细胞(BM-MNC)移植治疗肢体缺血,初步探讨SDF-1促进血管新生的机理和疗效。方法建立大鼠左后肢缺血模型,将动物随机分为非缺血对照组,缺血对照组,SDF-1局部应用组,自体BM-MNC移植组以及两者联合应用组。检测缺血后24 h左后肢腓肠肌SDF-1含量及CD133+细胞数量;于移植后4周动脉造影后处死动物,免疫组化法检测缺血区新生血管密度,评价血管新生情况。结果缺血对照组24 h肌组织SDF-1含量为(1.31±0.20)ng/50μg总蛋白,非缺血对照组为(0.93±0.29)ng/50μg总蛋白(P<0.05); CD133+细胞数(2.10±0.62)vs.(0.24±0.10)个/HP,P<0.01。SDF-1局部应用可增加缺血区CD133+细胞浸润数量(3.64±0.69)个/HP(P<0.01);并可提高四周后组织内新生血管数量;SDF-1与自体骨髓单个核细胞移植联合应用,可进一步提高新生血管数量。结论SDF-1可能在肢体缺血后自体干细胞的动员中起重要作用,其局部应用可促进缺血区血管新生或进一步提高干细胞移植治疗缺血的疗效。  相似文献   

11.
We investigated bone marrow mononuclear cells (BM-MNCs) and peripheral blood mononuclear cells (PB-MNCs) for therapeutic angiogenesis in the ischemic hindlimb. BM-MNCs were isolated and injected into ischemic skeletal muscles in mice. Laser Doppler and histological evaluation were performed after the surgical procedure. Fifteen patients suffering from critical lower limb ischemia received subcutaneous injections of recombinant human granulocyte colony-stimulating factor (G-CSF) to mobilize progenitor cells, and PB-MNCs were harvested and transplanted directly into the ischemic limb. Endothelial cells derived from BM-MNCs were plated, then induced to form three-dimensional networks by invading a Matrigel. Four weeks after implantation of BM-MNCs, laser Doppler analysis showed that the blood flow ratio was significantly increased (0.67 +/- 0.02 vs. 0.44 +/- 0.02). Alkaline phosphatase and immunohistochemical analyses showed that capillary density was significantly increased (95.25 +/- 0.07% vs. 39.6 +/- 0.04%). Two months after implantation of PB-MNCs, in both subgroups, ankle-brachial index values, walking distance, pain scale, and transcutaneous oxygen pressure (TcO(2)) were significantly improved (p < 0.005). A total of six of 15 limb ulcers of transplanted patients were healed after cell transplantation. BM-MNC implantation was able to induce functional angiogenesis in mice ischemic hindlimb. This clinical trial shows that G-CSF-based PB-MNC transplantation is a feasible treatment for the ischemic hindlimb.  相似文献   

12.
BACKGROUND: Bone marrow possesses endothelial progenitor cells that secrete several growth factors and can contribute to the formation of new capillaries. In the present study, we investigated the extent of angiogenesis induced by implantation of autologous bone marrow cells (BMCs) in a rat ischemic hindlimb model and studied whether the increased collateral vessels can improve deteriorated physical function. MATERIALS AND METHODS: Ischemic hindlimb was created by ligation of the femoral artery and its branches in Dark Agouti (DA) rats. BMCs (1 x 10(7)) were injected percutaneously at six points into the gastrocnemius muscle. To assess angiogenesis, histologic evaluation and microangiography were performed at 2 weeks postligation. Severity of the ischemic insult was evaluated by measuring blood flow in the adductor and gastrocnemius muscles using nonradioactive colored microspheres and by determining the femoral arteriovenous oxygen difference (AVDO(2)) at 2 weeks postligation. Running time on a motor-driven treadmill was used to represent exercise capacity. RESULTS: The histologic evaluation and microangiogram showed that the implanted BMCs induce angiogenesis. Blood flow to the adductor muscle on the treated side in the bone marrow cell implantation (BMI) group was significantly restored to 77.3 +/- 19.3% of that of the normally perfused limb in comparison to that in control groups (P < 0.05). AVDO(2) in the BMI group significantly decreased when compared with AVDO(2) in control groups. Rats in the BMI group ran approximately 1.5 times longer than rats in control groups at 2 and 4 weeks postligation (P < 0.01). CONCLUSIONS: Implantation of autologous BMCs induced angiogenesis and improved deteriorated exercise capacity in our rat ischemic hindlimb model.  相似文献   

13.
We hypothesized that in vitro treatment of peripheral blood mononuclear cells (PB-MNCs) from diabetic patients with ephrin-B2/Fc (EFNB2) improves their proangiogenic therapeutic potential in diabetic ischemic experimental models. Diabetes was induced in nude athymic mice by streptozotocin injections. At 9 weeks after hyperglycemia, 10(5) PB-MNCs from diabetic patients, pretreated by EFNB2, were intravenously injected in diabetic mice with hindlimb ischemia. Two weeks later, the postischemic neovascularization was evaluated. The mechanisms involved were investigated by flow cytometry analysis and in vitro cell biological assays. Paw skin blood flow, angiographic score, and capillary density were significantly increased in ischemic leg of diabetic mice receiving EFNB2-activated diabetic PB-MNCs versus those receiving nontreated diabetic PB-MNCs. EFNB2 bound to PB-MNCs and increased the adhesion and transmigration of PB-MNCs. Finally, EFNB2-activated PB-MNCs raised the number of circulating vascular progenitor cells in diabetic nude mice and increased the ability of endogenous bone marrow MNCs to differentiate into cells with endothelial phenotype and enhanced their proangiogenic potential. Therefore, EFNB2 treatment of PB-MNCs abrogates the diabetes-induced stem/progenitor cell dysfunction and opens a new avenue for the clinical development of an innovative and accessible strategy in diabetic patients with critical ischemic diseases.  相似文献   

14.
目的:探讨大鼠骨髓来源的血管内皮祖细胞(endothelial progenitor cells,EPCs)移植促进缺血皮瓣的血管新生,从而提高皮瓣存活率的可能性。方法:在Wistar大鼠背部形成2cm×8cm蒂部位于双侧髂棘连线上的随意型超比例皮瓣,建立缺血皮瓣模型。采用密度梯度离心法从Wistar大鼠骨髓中分离出骨髓单个核细胞,并在专有的诱导培养基(EG-M2MV)中培养,贴壁筛选法分离,诱导分化为EPCs;对细胞的形态、表型、功能加以鉴定。将EPCs注射移植于iWstar大鼠背部的缺血皮瓣,大体观察皮瓣的存活率,vWF免疫酶染色法检测皮瓣毛细血管密度。以缺血皮瓣单纯注射磷酸盐缓冲液(PBS)的Wistar大鼠作为对照组。结果:骨髓中分离培养的EPCs呈现典型的"铺路石"样外观,表达CD34、Flk-1及CDl 33表型,能够摄取DIL-ac-LDL和特异性结合FITC-UEA-1。注射EPCs组的缺血皮瓣存活率以及毛细血管密度显著高于对照组(P〈0.05)。结论:骨髓中的EPCs在体外培养后注射移植于皮瓣内,可以促进缺血性皮瓣的血管新生,提高缺血性皮瓣的存活率。  相似文献   

15.
We established a comparative model of angiogenic induction in previously formed fibrocollagenous tunnels in rat inner thigh muscles. A unilateral hindlimb chronic ischemia model was performed in male Sprague-Dawley rats. A device was then inserted in the central portion of the inner thigh muscles. Vascularity in the ischemic limb was determined by means of an angiographic score, capillary/fiber ratio, and endothelial proliferation by histochemistry and immunohistochemistry. Autologous transplant of bone marrow, vascular endothelial growth factor (VEGF), or collagen-polyvinylpyrrolidone plus heparin induced significant vascularization of the ischemic hindlimb when compared to saline solution. However, the bone marrow group presented a higher angiographic score than the other two. No differences among groups were observed in capillary/fiber ratio or proliferation, except for the VEGF group, where capillary proliferating cells were significantly higher than in controls. Based on these results, bone marrow-derived progenitor cells may constitute a safe and viable alternative for the induction of therapeutic angiogenesis.  相似文献   

16.
Implantation of autologous bone marrow (BM) mononuclear cells (MNCs) has been shown to augment neovascular formation in ischemic tissues in experimental animals and in humans. Prostaglandin derivatives improve the symptoms of patients with critical limb ischemia, possibly by their vasodilating and antiplatelet actions. We therefore examined whether therapeutic angiogenesis by implantation of autologous BM-MNCs would be enhanced by beraprost sodium (BPS), using a rabbit ischemic hindlimb model. Ischemia was induced by surgical resection of the left femoral artery. Twenty-five New Zealand white rabbits were divided into four groups. The first group (BM group, n = 4) received autologous BM-MNCs (2 × 106/animal) implanted into the ischemic tissue 1 week after limb ischemia. The second group (BM+BPS group, n = 8) received BPS injected into the dorsal skin (300 μg/kg daily) starting 1 week before limb surgery. This group received BM-MNC implantation 1 week after surgery. Daily injection of BPS was continued until the end of the protocol. The third group (BPS group, n = 8) received BPS injected into the dorsal skin (600 μg/kg daily) starting 1 week before limb surgery. The fourth group received saline as a control (n = 4). The extent of angiogenesis in the ischemic hindlimb was assessed using the angiographic score (AS), ischemic/normal limb calf blood pressure ratio (CBPR), and tissue capillary density. Four weeks after limb ischemia, the ischemic/normal CBPR was highest in the BM+BPS group, followed by the BPS, BM, and control groups (0.56 ± 0.16, 0.51 ± 0.25, 0.44 ± 0.15, and 0.30 ± 0.10, respectively). The AS was also the greatest in the BM+BP group, followed by the BM, BP, and S group (1.63 ±0.21, 1.31 ± 0.25, 1.26 ± 0.21 and 0.80 ± 0.10, respectively). The TCD was greatest in the BM+BP group, followed in by the BM, BP, and S group? (46 ± 4.1, 34 ± 0.7, 33 ± 6.9, and 19 ± 1.8 per field, respectively). BP treatment is an effective means to enhance the efficacy of therapeutic angiogenesis induced by autologous BM-MNCs implantation in ischemic hindlimb tissues.  相似文献   

17.
BACKGROUND: Therapeutic angiogenesis was induced by local autologous bone marrow cell implantation (BMCI) in ischemic hindlimb or ischemic heart models in rats. This study was designed to investigate the toxicity and therapeutic potency of local BMCI using a chronic coronary occlusion model in dogs. METHODS: The canine chronic coronary occlusion model was created by ligating of the left anterior descending artery (LAD). The myocardium in the left ventricle was divided into distinct normal, marginal, and infarction areas 30 days after LAD ligation. Each area was injected at two locations, with either 2 x 10(7) bone marrow cells (n = 7, BMCI group) or 0.1 mL phosphate-buffered saline (PBS) only (n = 7, PBS group), respectively. Hemodynamics were evaluated by a single ultrasonic transducer and echocardiography before and 30 days after the treatment. Angiogenesis was evaluated by vessel count 30 days after the treatment. The toxicity of BMCI treatment was also evaluated in 8 normal dogs by following changes in electrocardiography (ECG), echocardiography, local histology, and systemic biochemistry indexes. RESULTS: There was a significantly higher percentage of wall thickening in the marginal area in the BMCI group than in the PBS group 30 days after treatment (14.5 +/- 2.28 versus 8.1 +/- 3.00, p = 0.002). Significantly more microvessels were observed in the marginal area in the BMCI group than in the PBS group 30 days after treatment (127.7 +/- 20.1 versus 88.0 +/- 10.2/field, p = 0.0007). No systemic or local toxicity was found following BMCI treatment in the acute or chronic phases. CONCLUSIONS: BMCI treatment improved local wall thickening dynamics, presumably due to the angiogenesis induced by the treatment. This indicates that it might be a safe and effective therapy for ischemic heart disease.  相似文献   

18.
目的观察骨髓动员刺激后自体骨髓单个核细胞移植治疗下肢缺血的初步疗效。方法2005年5月~2005年12月收治下肢缺血35例43侧患肢,男23例,女12例。年龄34~90岁,平均71.3岁。病因:糖尿病下肢缺血30例38侧患肢,单纯动脉硬化闭塞症2例2侧患肢,血栓闭塞性脉管炎3例3侧患肢。其中间歇性跛行期5例5侧患肢;静息痛期15例19侧患肢;组织缺损期15例19侧患肢,其中组织溃疡期9例12侧患肢;组织坏疽期6例7侧患肢。在抽取骨髓前使用粒细胞集落刺激因子刺激骨髓2~3d,每天300μg;抽取骨髓血130-200ml,经分离纯化后再行移植。采用下肢肌肉局部注射13例19侧患肢,下肢动脉腔内注射16例16侧患肢,下肢肌肉局部注射与动脉腔内同时注射6例8侧患肢进行移植。结果术后2个月肢体疼痛改善率为94.7%,冷感改善率为97.1%,肢体麻木改善率为93.3%。5例5侧患肢间歇性跛行距离均有不同程度增加。44.2%患者的踝肱比值(ankle/brachial index ABI)有不同程度增加。39侧患肢行经皮氧分压(transeutaneous oxygen pressure,TeP02)测定,92.3%有不同程度增高。患肢溃疡:在9例11侧患肢中,愈合1侧,明显缩小或缩小3侧,无变化7侧,其中3侧患肢被截肢。术后行血管造影评估25例34侧患肢,91.2%患肢的侧支循环有不同程度增加。并发症:骨髓动员刺激出现发热和轻微乏力各1例,均自行缓解;单个核细胞移植后1周出现轻度心肌梗死1例,药物治疗1周后恢复出院,1个月后因患肢疼痛加重行膝下截肢。32例40侧患肢获随访3412个月,症状消失13侧,明显改善15侧,改善8侧,复发2侧,无效2侧。客观评价标准与术前比较ABI增加25侧;TcPOz测定增加36侧;21侧患肢的血管造影显示90.5%患肢有新生侧支形成;10侧患肢足部溃疡7侧愈合,3侧明显缩小;3侧患肢截除坏疽足趾者于术后2~3个月愈合出院。结论经骨髓动员刺激后的骨髓单个核细胞移植下肢缺血,具有抽取骨髓血少、细胞量多、近期效果好且安全性高的优点,是除自体骨髓单个核细胞移植和外周血干细胞移植外的又一种治疗下肢缺血的新方法。远期效果尚需进一步随访。  相似文献   

19.
目的 比较骨髓单个核细胞(MNCs)和骨髓源内皮祖细胞(EPCs)移植促进血流重建的效果,探讨非内皮祖细胞在血流重建中的作用.方法 获取Lewis大鼠骨髓MNCs,部分MNCs在体外诱导分化为EPCs.采用Lewis大鼠建立单侧后肢缺血模型.建模后3 d,将模型鼠随机分为3组:(1)对照组(n=6),将0.8 mL D-Hank's液注入对照组大鼠缺血侧后肢;(2)MNC组(n=6),将8×10~6个骨髓MNC植入MNC组大鼠缺血侧后肢;(3)EPC组(n=6),将体外培养的8 × 10~6个EPC植入EPC组大鼠缺血侧后肢.细胞移植后3周行大鼠后肢动脉造影,检测缺血侧后肢侧支血管数;切取缺血侧后肢腓肠肌,分别行CD31和α-SMA免疫组化染色,计算毛细血管密度和小动脉密度.结果 MNC组毛细血管密度与EPC组差异无统计学意义[(31.67 ± 7.87)个/HP vs.(32.83±5.38)个/HP,P>0.05].而均高于对照组(19.67 ± 4.80个/HP)(P<0.05);MNC组侧支血管数与EPC组差异无统计学意义[(4.17±0.75)个vs.(4.50 4±1.38)个,P>0.05],但均高于对照组[(2.50 ± 1.5)个](P<0.05);MNC组小动脉密度与EPC组差异无统计学意义[(4.83 ± 1.47)个vs.(5.50 ± 2.35)个,P>0.05],亦均高于对照组[(2.17±0.98)个](P<0.05).结论 在骨髓干细胞移植治疗肢体缺血性疾病中,非内皮祖细胞在血流重建中所起的作用与EPC相似.  相似文献   

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