Autologous stem cell transplantation in acute myocardial infarction: The ASTAMI randomized controlled trial. Intracoronary transplantation of autologous mononuclear bone marrow cells, study design and safety aspects

OBJECTIVES: Intracoronary transplantation of different cell populations has been used in acute myocardial infarction (AMI) with promising results. The primary objective of the Autologous Stem cell Transplantation in Acute Myocardial Infarction (ASTAMI) study is to test whether intracoronary transplantation of autologous mononuclear bone marrow cells (mBMC) improves left ventricular ejection fraction (LVEF) after anterior wall AMI. DESIGN: The ASTAMI study is a randomized, controlled, prospective study. One hundred patients with acute anterior wall ST-elevation myocardial infarction (STEMI) treated with acute percutaneous coronary intervention (PCI) are randomized in a 1:1 way to either intracoronary transplantation of autologous mBMC 5-8 d after PCI or to control. Left ventricular function, exercise capacity, biochemical status, functional class, quality of life and complications are validated at baseline and during a 12-month follow-up. RESULTS: By August 2004, out of 1004 patients with STEMI, 49 patients have been included in the study. Twenty-four patients have been randomized to intracoronary mBMC transplantation. Twenty patients had chest pain and 16 patients had ischemic ECG changes during the mBMC transplantation procedure. One patient had ventricular fibrillation 24 h after transplantation. CONCLUSIONS: Intracoronary transplantation of autologous mBMC in the acute phase after AMI is feasible and seems safe in the short term.     

骨髓单个核细胞移植治疗急性心肌梗死

目的探讨自体骨髓单个核细胞(BMMNCs)移植对急性心肌梗死后心脏功能及结构的影响。方法中国小型猪16头,随机分为对照组(n=6)及移植组(n=10)。建立急性心肌梗死模型后,分别注入BMMNCs/生理盐水。两组动物于术中监测血流动力学指标,术后进行影像学检查。移植组分别于2、4周后获取心脏标本,检测移植细胞标记物Hoechst33342。结果心梗4周后,移植组心梗范围较对照组显著缩小(P<0.01);血流动力学指标较对照组明显改善;局部室壁运动评分优于对照组,组间差异有统计学意义(P<0.05)。在术后2周获得的心肌标本内,可见多量Hoechst33342和MHC双阳性细胞。结论成体BMMNCs移植后,有可能抑止心室重构的发展,提高心室运动功能。     

自体骨髓单个核细胞移植治疗下肢慢性静脉溃疡的初步研究

目的探讨自体骨髓单个核细胞移植治疗下肢慢性静脉溃疡的疗效。方法 2009年5月-2010年9月,在采用大隐静脉剥脱术基础上,对17例下肢慢性静脉溃疡患者采用自体骨髓单个核细胞移植治疗(移植组),并与同期未采用细胞移植治疗的10例患者(对照组)比较疗效。移植组:男9例,女8例;年龄(33.3±6.1)岁。单纯大隐静脉曲张慢性溃疡11例,单纯深静脉瓣膜功能不全慢性溃疡6例。溃疡面积(4.39±2.46)cm2。病程3个月～6年。对照组:男4例,女6例;年龄(39.2±10.3)岁。单纯大隐静脉曲张慢性溃疡7例,单纯深静脉瓣膜功能不全慢性溃疡3例。溃疡面积(5.51±2.63)cm2。病程3个月～2年。两组按照临床病因解剖病理学分级(CEAP)均为C6级。两组患者一般资料比较差异均无统计学意义(P>0.05),有可比性。观察术后溃疡愈合情况;移植组于术前及术后3 d分别切取溃疡面肉芽组织行常规HE染色,免疫组织化学染色观察VEGF表达和微血管密度(microvessel density,MVD)。结果移植组患者溃疡愈合较快,其中15例愈合,1例好转,1例未愈合,中位愈合时间为22 d,四分位数间距为15 d;对照组溃疡愈合缓慢,其中7例愈合,3例未愈合,中位愈合时间57.5 d,四分位数间距为40 d。两组溃疡愈合时间比较差异有统计学意义(Z=0.001 4,P=0.002 7)。移植组HE染色显示,细胞移植后溃疡肉芽组织有丰富毛细血管结构;免疫组织化学染色示移植后MVD为(32.1±12.8)个,较移植前(22.1±6.7)个显著增加,差异有统计学意义(t=3.120,P=0.008);移植后VEGF表达阳性细胞百分比为8.05%±5.10%,较移植前(6.13%±4.20%)升高,但差异无统计学意义(t=1.150,P=0.268)。结论自体骨髓单个核细胞移植可以促进下肢慢性静脉溃疡肉芽组织增生,加速溃疡愈合。     

3种不同途径移植自体骨髓间充质干细胞治疗急性心肌梗死效果比较

目的比较3种不同途径移植自体骨髓间充质干细胞(MSCs)治疗急性心肌梗死效果。方法小型猪12只,建立心肌梗死(AMI)模型,纯化扩增并将DAPI标记的MSCs经冠脉、心内膜及心外膜注射移植,移植前后记录左室血流动力学指标及LVEF变化,3个月后取心肌行免疫组织化学检测结蛋白desmin和心肌特异性肌钙蛋白I(cTnI)的表达。结果冠脉组梗死心肌周围未见明确的移植细胞,仅见非特异的荧光染色,与心梗后比较心功能改善,但与对照组比较无明显差异;心内膜及心外膜组移植的MSCs分布较广,胞核为蓝色椭圆形,免疫组织化学检测胞浆心肌特异蛋白染色阳性,与冠脉移植组比较LVEDP降低(P<0．05)、LV±dp／dtmax增快(P<0．05)、 LVEF增高(P<0．05)、新生血管数增加及瘢痕面积缩小等指标改善更明显,且有统计学意义。结论 3种途径移植MSCs均可改善AMI后心功能,经冠脉移植不是最佳途径,心肌内注射效果更好。     

自体外周血及自体红骨髓干细胞与脱钙骨复合移植治疗骨缺损的比较研究

目的 对比观察自体外周血干细胞（APBSC）／脱钙骨（DB）复合移植与自体外红骨髓（ARBM）／DB复合移植治疗骨缺损的疗效。方法 36只家兔双侧桡骨造成1cm骨缺损,随机分为DB组、ARBM／DM级和APBSC／DB组,每组12只,分别进行X线片、生物力学和组织学检查,然后作对比分析。结果 术后第2、4、8、14周,APBSC／DB组和ARBM／DB组X线片、改进的GaryX线评分和光镜观察结果,以及术后第14周整骨破坏载荷和骨缺损修复形态学评分均明显优于DB组;但APBSC／DB组与ARBM／DB组间差异无显著性。结论 APBSC与ARBM都能在DB的骨形态蛋白诱导下促进成骨细胞的形成,复合移植疗效明显优于单纯DB移植。     

The effect of magnesium supplements on early post‐transplantation glucose metabolism: a randomized controlled trial

Post‐transplantation hypomagnesemia is common and predicts diabetes. Magnesium improves glycemic control in diabetics and insulin sensitivity in insulin resistant subjects. We aimed to assess the effectiveness of oral magnesium for improving glycemic control and insulin sensitivity at 3 months post‐transplantation. We conducted a single‐center, open‐label, randomized parallel group study. We included adults with serum magnesium <1.7 mg/dl within 2 weeks after kidney transplantation. We randomized participants to 450 mg magnesium oxide up to three times daily or no treatment. The primary endpoint was the mean difference in fasting glycemia. Secondary endpoints were the mean difference in area under the curve (AUC) of glucose during an oral glucose tolerance test and insulin resistance measured by Homeostasis Model of Assessment‐Insulin Resistance (HOMA‐IR). Analyses were on intention‐to‐treat basis. In patients randomized to magnesium oxide (N = 27) versus no treatment (N = 27), fasting glycemia on average was 11.5 mg/dl lower (95% CI 1.7 to 21.3; P = 0.02). There was no difference between the two groups neither for 2 h AUC, where the mean value was 1164 mg/dl/min (95% CI ?1884 to 4284; P = 0.45) lower in the treatment group nor for HOMA‐IR. Magnesium supplements modestly improved fasting glycemia without effect on insulin resistance. Higher baseline glycemia among patients in the control group may have driven the positive outcome (ClinicalTrials.gov number: NCT01889576).     

由胚胎干细胞分化的心肌细胞移植对心肌梗死大鼠左心室重构及心功能的影响

目的探讨小鼠胚胎干细胞（ES）分化的心肌细胞移植对SD大鼠心肌梗死后左心室重构及心功能的影响。方法采用悬滴-悬浮培养法诱导ES分化形成拟胚体。其中包含有大量心肌细胞。将雄性SD大鼠冠状动脉左前降支结扎。制成急性心肌梗死模型。1周后再次开胸。实验组于梗死区域注入心肌细胞悬液150μl（含细胞5×10^5个），梗死对照组于梗死区域注入等体积细胞培养液，另设假手术组，不结扎冠状动脉。心肌梗死后1周和细胞移植后4周，采用超声心动图评价大鼠左心室形态和功能，用免疫荧光技术检测移植细胞的存活及其表型。结果免疫荧光技术检测到分化的心肌细胞表达特异性转录因子α辅肌动蛋白、α肌球蛋白重链和肌钙蛋白1；逆转录聚合酶链反应检测到有心肌特异性转录因子Nkx-2．5、GATA-4及α肌球蛋白重链基因的表达；膜片钳记录显示所得到的分化的心肌细胞包括窦房结样起搏细胞、心房样细胞和心室样细胞。细胞移植后4周。与梗死对照组相比，实验组左心室收缩末期容积和舒张末期容积均显著缩小，左心室前壁舒张末期厚度明显增加（P〈0．01）。左室长轴缩短率显著升高（P〈0．01）。心脏组织冰冻切片可见4,6-联脒-2-苯基吲哚标记的呈蓝色荧光的移植细胞核。移植细胞表达肌钙蛋白I，移植细胞与受者的心肌细胞间有连接蛋白-43的表达。实验组中有2只（12．5％。2／16）出现了心脏畸胎瘤。结论由胚胎干细胞分化的心肌细胞移植可逆转心肌梗死大鼠左心室重构。改善左心室功能。     

骨髓间质干细胞移植对香猪急性心肌梗死后心肌结构和心功能的影响

目的 移植自体骨髓间质干细胞(BMSCs)到香猪急性心肌梗死区内,研究移植BMSCs对心肌结构和心功能的影响. 方法将24只贵州香猪采用计算器随机法分为实验组(n＝12)和对照组(n＝12),抽取香猪自体骨髓,经体外分离出BMSCs并培养和经5-氮胞苷(5-azacytidine)转化,利用结扎左前降支(LAD)的方法建立急性心肌梗死动物模型,经LAD和梗死区多点注射的方法将实验组香猪注射BMSCs(细胞总数2×106个),对照组注射等量的细胞培养液.3周和6周后,用超声心动图(UCG)观察两组移植后心肌结构和心功能改变的情况. 结果实验组左心室射血分数、左心室短轴缩短率和室壁增厚率明显高于对照组;左心室室壁、室间隔厚度和心室腔的大小在两组之间也存在明显差别,实验组室壁和室间隔厚度明显大于对照组,而心室腔小于对照组. 结论 BMSCs梗死区心肌移植后可减轻心室重构的进程,减轻心肌的变薄程度,使心室腔未明显扩大.BMSCs移植还可增加心肌的收缩力,改善心功能.     

大鼠骨髓间充质细胞经SDF-1预处理后移植对急性心肌梗死的治疗效果

目的 探讨大鼠骨髓间充质细胞(BMSC)经基质细胞衍生因子-1(SDF-1)预处理后移植对急性心肌梗死(AMI)的治疗效果.方法 (1)全骨髓贴壁法培养大鼠BMSC;逆转录聚合酶链反应(RT-PCR)和免疫组织化学法检测BMSC趋化因子受体(CXCR4)基因的表达;将BMSC分别与10和100μg/L的SDF-1作用24 h后,在无氧、无血清条件下培养6 h,流式细胞仪和末端标记(TUNEL)法检测细胞凋亡率.(2)建立大鼠急性心肌梗死模型,将经100μg/L SDF-1预处理的BMSC和未经处理的BMSC植入大鼠梗死心肌周边,2周后采用超声心动图观察心脏功能的变化.结果 BMSC表达CXCR4基因;在无氧和无血清条件下,经SDF-1预处理的BMSC凋亡率与对照组比较明显降低(P<0.05),而经100 μg/L SDF-1预处理的BMSC凋亡率最低.成功建立大鼠急性心肌梗死模型;与未经处理的BMSC移植比较,经SDF-1预处理的BMSC移植后改善心肌梗死大鼠的心功能作用更为明显(P<0.05).结论 用SDF-1预处理大鼠BMSC能抑制其在无氧和无血清条件下的凋亡.用SDF-1预处理BMSC能增强其移植后治疗大鼠急性心肌梗死的效果.     

自体骨髓干细胞移植治疗激素性股骨头坏死

目的观察自体骨髓干细胞移植治疗激素性股骨头坏死的临床效果。方法52例激素性股骨头缺血性坏死患者经使用rhG-CSF进行骨髓干细胞动员后,第2天进行骨髓干细胞采集,在数字减影血管造影术（DSA）下行股动脉穿刺,导管超选择至闭孔动脉及旋股内外侧动脉,将干细胞悬液缓慢匀速注入动脉内;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,行走间距及步态变化,髋关节外展及内旋功能变化;6个月后行股骨头供血动脉造影术。结果52例患者随访12个月,髋关节疼痛有不同程度缓解67％,关节功能改善29．7％,行走间距延长32．5％。干细胞移植术后6个月,10例患者行股骨头供血动脉数字减影血管造影检查,均显示旋股内动脉,旋股外动脉及闭孔动脉管径增粗,新生血管增多,血流速度增快,与移植前血管造影相比较,股骨头血液供应明显改善。治疗中均未发生严重的并发症和不良反应。结论自体骨髓干细胞移植治疗激素性股骨头坏死方法简便,安全有效,适合Ⅰ-Ⅲ期早、中期股骨头坏死的患者。     

双份脐血移植后受者骨髓间充质干细胞嵌合情况

目的 研究双份脐血移植(DCBT)受者骨髓间充质干细胞(MSC)的嵌合状态.方法 急性粒细胞白血病M2a型男性患者1例,接受改良白消安环磷酰胺方案+抗胸腺细胞球蛋白(ATG)预处理,输注5个抗原(5/6)相合和4个抗原(4/6)相合的非血源脐血各1份,移植后19 d粒系造血重建.移植后87 d,采用密度梯度离心法分离DCBT后受者及正常供者的骨髓单个核细胞,分别培养MSC,用流式细胞术检测细胞表面标志,诱导其向成脂肪细胞和成骨细胞分化,应用逆转录聚合酶链反应法检测MSC表面造血及免疫相关分子的表达,短串联重复序列聚合酶链反应检测受者MSC、外周血、骨髓中供者细胞嵌合率.结果 移植后受者MSC与正常供者MSC具有相似的细胞形态、免疫表型以及分化潜能,均能表达白细胞介素6、干细胞因子、白血病抑制因子和粒-巨噬细胞集落刺激因子等造血及免疫相关分子的mRNA.DCBT后受者骨髓优势脐血嵌合度达96.4％,外周血嵌合度达95.7％,MSC的优势脐血嵌合度为5.4％,MSC中受者本身部分占94.6％.结论 DCBT后,受者造血重建仅来自于其中1份脐血.移植后骨髓MSC大部分来源受者本身,部分嵌合的供者MSC来源于植入的单份脐血.     

骨髓间充质干细胞自体移植时间对兔急性肾功能衰竭治疗效果的影响

目的 观察骨髓间充质干细胞(MSCs)自体移植对兔急性肾功能衰竭的治疗作用并探讨不同移植时间对其治疗效果的影响.方法 骨髓穿刺抽取新西兰大耳白兔骨髓,分离、培养、扩增MSCs.30只兔通过夹闭双肾动脉90 min后再灌注制作急性肾功能衰竭模型,随机分为移植A组、移植B组和未治疗C组,每组10只.移植A组在恢复血流即刻,移植B组在恢复血流后72 h将BrdU标记的自体MSCs经颈静脉回输移植,未治疗C组仅制作急性肾功能衰竭模型.于造模后21 d处死兔,比较各组的存活率、肾功能及肾组织形态学改变.结果 与未治疗组比较,移植组肾功能较快恢复(P<0.05),肾组织损伤明显改善(P<0.05).移植A组的效果优于移植B组(P<0.05).结论 骨髓间充质干细胞自体移植能有效治疗缺血再灌注损伤引起的急性肾功能衰竭,早期移植的效果好.     

Bone marrow-derived cell transplantation therapy for myocardial infarction: lessons learned and future questions

Over the last decade, many investigators have utilized bone marrow-derived cells for cell transplantation therapy in animal studies and in patients with acute myocardial infarction and chronic heart failure. In those experimental and clinical studies, various doses and types of bone marrow-derived cells have been transplanted to the injured myocardium using a variety of approaches, such as intracoronary infusion or catheter-based direct endomyocardial injection, and at different time points after successful coronary reperfusion. The reported treatment effects are variable, which may be related to differences in cell type and quantity of transplanted cells, timing and approach of cell transplantation and patient selection. In this review, we summarize and discuss the controversies and questions related to the clinical use of bone marrow-derived cells.     

The effect of bright light therapy on sleep and circadian rhythms in renal transplant recipients: a pilot randomized,multicentre wait‐list controlled trial

This study assessed the effect and feasibility of morning bright light therapy (BLT) on sleep, circadian rhythms, subjective feelings, depressive symptomatology and cognition in renal transplant recipients (RTx) diagnosed with sleep–wake disturbances (SWD). This pilot randomized multicentre wait‐list controlled trial included 30 home‐dwelling RTx randomly assigned 1:1 to either 3 weeks of BLT or a wait‐list control group. Morning BLT (10 000 lux) was individually scheduled for 30 min daily for 3 weeks. Wrist actimetry (measuring sleep and circadian rhythms), validated instruments (subjective feelings and cognition) and melatonin assay (circadian timing) were used. Data were analysed via a random‐intercept regression model. Of 30 RTx recipients (aged 58 ± 15, transplanted 15 ± 6 years ago), 26 completed the study. While BLT had no significant effect on circadian and sleep measures, sleep timing improved significantly. The intervention group showed a significant get‐up time phase advance from baseline to intervention (+24 min) [(standardized estimates (SE): ?0.23 (?0.42; ?0.03)] and a small (+14 min) but significant bedtime phase advance from intervention to follow‐up (SE: ?0.25 (?0.41; ?0.09). Improvement in subjective feelings and depressive symptomatology was observed but was not statistically significant. Bright light therapy showed preliminary indications of a beneficial effect in RTx with sleep–wake disturbances. (ClinicalTrials.gov number: NCT01256983)     

Early results of core decompression and autologous bone marrow mononuclear cells instillation in femoral head osteonecrosis: a randomized control study

Fifty-one osteonecrotic hips in 40 patients were randomly divided into 2 treatment groups. Patients in group A (25 hips) were treated with core decompression, and those in group B (26 hips) received autologous bone marrow mononuclear cell instillation into the core tract after core decompression. Outcome between the 2 groups were compared clinically (Harris Hip score), radiographically (x-ray and magnetic resonance imaging), and by Kaplan-Meier hip survival analysis after 12 and 24 months of surgical intervention. The clinical score and mean hip survival were significantly better in group B than in group A (P < .05). Patients with adverse prognostic features at initial presentation, that is, poor Harris Hip score, x-ray changes, edema, and/or effusion on magnetic resonance imaging had significantly better clinical outcome and hip survival in group B than in group A.     

咪达唑仑和异丙酚对急性心肌梗死大鼠血清VEGF浓度及G-CSF药物动员骨髓干细胞效果的影响

目的 评价咪达唑仑和异丙酚对急性心肌梗死大鼠血清血管内皮生长因子(VEGF)浓度及粒细胞集落刺激因子(G-CSF)药物动员骨髓干细胞效果的影响.方法 雄性Wistar大鼠36只,体重250 ～ 280 g,采用结扎左冠状动脉前降支的方法制备急性心肌梗死模型,采用腹腔连续注射G-CSF 5d进行药物动员,于药物动员后第7天,按照随机数字表法,将大鼠随机分为G-CSF组(G组)、咪达唑仑组(M组)及异丙酚组(P组),每组12只.G组以0.5 ml/h的速率股静脉输注生理盐水6h;M组股静脉输注咪达唑仑0.05 mg·kg-·h-1 6 h;P组股静脉输注异丙酚5mg·kg-1 ·h-16 h.于给药完毕后经股静脉取血,采用流式细胞仅测定CD34+单核细胞( CD34+ MNC)和内皮祖细胞(EPCs)数目,采用ELISA法测定血清VEGF浓度.于心肌梗死后4周每组随机取6只大鼠测定左心室舒张末压(LVEDP)、最大收缩速率(+dp/dtmax)和最大舒张速率(- dp/max).结果 与G组比较,M组CD34+MNC及EPCs细胞数目增加,血清VEGF浓度升高,LVEDP下降,-dp/dtmax的绝对值升高(P＜0.05),P组LVEDP下降,- dp/dtmax的绝对值升高(P＜0.05);与P组比较,M组CD34+MNC及EPCs细胞数目增加,血清VEGF浓度升高,LVEDP下降,- dp/dtmax的绝对值升高(P＜0.05).结论 咪达唑仑可促进VEGF的释放,加强G-CSF动员骨髓干细胞的作用,改善急性心肌梗死后大鼠的心脏功能;异丙酚不能促进VEGF的释放及无骨髓干细胞动员的作用.     

Cx43转基因自体成肌细胞移植对犬急性心肌梗死后心肌结构和心功能的影响

目的移植Cx43转基因自体成肌细胞(Cx43 SMC)到犬急性心肌梗死(AMI)区内,观察移植Cx43 SMC对心肌结构和心功能的影响。方法将24条健康杂种犬采用计算器随机法分为3组(n=8),即成肌细胞(SMC)移植组、Cx43 SMC移植组和对照组。取犬自体骨骼肌,经体外分离、培养和转化,将质粒载体pLenti6/V5-DEST-Cx43转入骨骼肌成肌细胞。利用结扎左前降支(LAD)的方法建立AMI动物模型,经LAD和梗死区多点注射的方法将实验组犬注射SMC或Cx43 SMC(细胞总数2×10~6个),对照组注射等量的细胞培养液。分别在AMI前1d、AMI后4周、细胞移植4周后,用超声心动图(UCG)测量左室射血分数(LVEF)、左室舒张末期内径(LVDD)和左室收缩末期内径(LVSD)；并进行心肌组织的光镜和电镜的病理学检查。结果AMI前UCG检查犬左室形态和收缩功能正常,结扎LAD 4周后,LVEF降低、LVDD和LVSD均增加(P＜0．05)。移植后4周,与对照组和移植前比较,SMC和Cx43 SMC移植组的LVEF升高、LVDD和LVSD减少(P＜0．05),其中Cx43 SMC移植组较SMC移植组为显著(P＜0．05)。组织病理学检查亦显示移入的Cx43 SMC在宿主心肌组织中排列有序,与宿主心肌细胞的排列方向一致,其间有闰盘形成。结论Cx43 SMC梗死区心肌移植后可减轻心室重构的进程,增加心肌的收缩力,改善心功能。     

Autologous bone marrow-derived mesenchymal stromal cell therapy with early tacrolimus withdrawal: The randomized prospective,single-center,open-label TRITON study

After renal transplantation, there is a need for immunosuppressive regimens which effectively prevent allograft rejection, while preserving renal function and minimizing side effects. From this perspective, mesenchymal stromal cell (MSC) therapy is of interest. In this randomized prospective, single-center, open-label trial, we compared MSCs infused 6 and 7 weeks after renal transplantation and early tacrolimus withdrawal with a control tacrolimus group. Primary end point was quantitative evaluation of interstitial fibrosis in protocol biopsies at 4 and 24 weeks posttransplant. Secondary end points included acute rejection, graft loss, death, renal function, adverse events, and immunological responses. Seventy patients were randomly assigned of which 57 patients were included in the final analysis (29 MSC; 28 controls). Quantitative progression of fibrosis failed to show benefit in the MSC group and GFR remained stable in both groups. One acute rejection was documented (MSC group), while subclinical rejection in week 24 protocol biopsies occurred in seven patients (four MSC; three controls). In the MSC group, regulatory T cell numbers were significantly higher compared to controls (p = .014, week 24). In conclusion, early tacrolimus withdrawal with MSC therapy was safe and feasible without increased rejection and with preserved renal function. MSC therapy is a potentially useful approach after renal transplantation.