Construction of cardiac atrioventricular electrical conduction pathway by rabbit bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.41.007

Abstract

Abstract:

BACKGROUND: Autologous stem cell transplantation to the heart is a research direction of heart failure treatment, but there are relatively few of studies about autologous bone marrow mesenchymal stem cell transplantation targeting the cardiac conduction system.

OBJECTIVE: To evaluate the potential of rabbit bone marrow mesenchymal stem cells for treatment of heart block.

METHODS: Rabbit bone marrow mesenchymal stem cells were induced by 5-azacytidine to differentiate into cardiomyocyte-like cells. After thoracotomy, the left atrium-left ventricular anterior wall was sutured in 14 New Zealand white rabbits (8 in the experimental group and 6 in the control group). One month after the surgery, in the experimental group, autologous bone marrow mesenchymal stem cells induced by 5-azacytidine for 4 weeks were labeled with 4',6-diamidino-2-phenylindole and then injected into the suture area when opening the thoracic again. In the control group, cells cultured in medium were used. One month after cell transplantation, the third thoracotomy was done to insert electrodes into the left atrium and left ventricular anterior wall, for cardiac electrophysiological detection. Whether atrioventricular pathway formed in the suture area was observed.

RESULTS AND CONCLUSION: After cells were transplanted into the sutured area, two rabbits in the experimental group were discovered to form the atrioventricular pathway in the sutured area through cardiac electrophysiological examination. After transplantation, transplanted cells were visible on the heart frozen sections under fluorescence microscope in the left ventricle and sutured area, but there was no cell in the control group. In the experimental group, bone marrow mesenchymal stem cells expressed Cx43 and formed gap junction intercellular communication with cardiomyocytes, which was presented as formation of the atrioventricular pathway on cardiac electrophysiology examination. These findings indicate that bone marrow mesenchymal stem cells can be used to treat cardiac conduction system block diseases.

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

全文链接：

Key words: bone marrow, mesenchymal stem cell transplantation, cardiac electrophysiology

CLC Number:

R394.2

Zhou Hao-yue, Lu Jiong-bin, Qiu Han-ying. Construction of cardiac atrioventricular electrical conduction pathway by rabbit bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(41): 6597-6602.

Figures/Tables 2

2.1 实验动物数量分析参加实验兔数量总共为14只，其中实验组8只，对照组6只，其中实验组1只兔子在移植细胞后，心脏电生理检测术前麻醉时死亡，对照组1只兔子在行开胸手术，左房左室缝合时意外死亡。其余兔子存活至整个实验结束，最终实验组有7只，对照组有5只兔子进入最后统计分析。 2.2 房室缝合区组织苏木精-伊红染色和苦味酸-天狼星红染色实验组心肌标本苏木精-伊红染色结果提示，普通光镜下不能分辨正常心肌细胞与骨髓间充质干细胞分化形成的心肌样细胞，在左心房左心室缝合形成的瘢痕连接处，观察到有少量肌组织样结构形成。苦味酸-天狼星红染色后，正常心肌织呈淡黄色表现，而纤维结缔组织呈胭脂红样染色[23]。房室缝合处，形成纤维结缔组织，粘连较为紧密，可以成为细胞移植的平台，见图3A。在实验组中，可见散在、团状或断续条索淡黄色组织夹杂于纤维组织中，对照组无上述表现，但是值得注意的是，单纯的天狼星红染色不能确定有无心肌组织在瘢痕组织中形成，见图3B。 2.3 观察经5-氮胞苷诱导后的骨髓间充质干细胞在移植后的存活情况经荧光显微镜观察实验组细胞移植后存活情况，发现移植于正常心房与心室肌处的细胞存活较多，细胞分化好，苏木精-伊红染色下与正常心肌细胞难以区别，见图4A；而移植左房左室缝合部位的细胞能够在房室缝合处的瘢痕组织存活，但散在分布，呈细胞团或细胞索，细胞存活数量相对较稀少，明显少于接种于正常心肌处，见图4B。 2.4 移植前后体表常规心电图检测结果实验组细胞移植前后常规心电图各项指标观察差异无显著性意义(表1)，术前所有兔电生理检测未发现旁道(P > 0.05)。两组心率、PR间期、ORS间期在体表心电图上差异亦无显著性意义 (P > 0.05)，常规体表肢导联也未发现detla波(表2)。 2.5 房室旁道心脏电生理检测结果见图5。经分级递增起搏法和连续递增超搏法检测发现，在实验组中，有2只兔子采用分级递增起搏法刺激心房时，在起搏阀值为 0.6 mV，S1-S1间期为190 ms时，S1-R间期为98 ms，ORS间期为52 ms，S1-R间期随着S1-S1间期缩短而逐渐延长，见图5B。但是当S1-S1间期缩短至135-140 ms时，S1-R间期出现明显缩短伴随QRS波变化，S1-R间期从98 ms缩短为41 ms，而QRS波出现Q波明显缩小，时程延长至60 ms (这些兔在未实行刺激时，PR间期约为87 ms，QRS约为53 ms)，见图5C。但未能观察到类似人类心电图的典型detla波出现；在S1-S2刺激模式下，上述实验组中S1-R间期缩短的兔出现S2期前刺激后短阵的阵发性室上性心动过速表现，且R-P’间期短于PR间期，见图5D；而对照组兔皆未出现上述实验结果，未发现有S1-R间期缩短及QRS波群变化，亦未观察到detla波出现。所有兔以同样的程序刺激左心室，未能观察到隐匿性旁道的存在。上述实验组中出现电生理刺激改变的兔子同样未能检测到旁道逆传现象。"

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