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目的探讨超声微泡造影剂介导对猪骨髓干细胞(BMSC)心肌内移植归巢的影响。方法选取8只2月龄中国家猪随机分为对照组(A组,4只)和实验组(B组,4只)。分别抽取骨髓,体外分离、纯化,培养BMSC,超顺磁性氧化铁颗粒(SPIO)标记;介入法建立急性心肌梗死模型。B组超声微泡作用下移植BMSC(1MHz,2W/cm2,90s辐射);A组不采用超声微泡干预BMSC移植过程。两组动物于术后24h活体行磁共振检查,对比BMSC移植,随后处死动物,取心肌梗死区、梗死周边区、正常区做病理切片,行HE染色及普鲁士蓝染色,心肌消化后涂片普鲁士蓝染色后计数BMSC。结果①SPIO成功标记干细胞,因此,活体行磁共振示踪BMSC成为可能。②磁共振:A组及B组在心肌梗死区和梗死周边区均见SPIO标记的BMSC形成的低密度区,低信号区B组较A组多。③普鲁士蓝染色:两组心肌消化涂片染色,BMSC在B组较A组有明显地增加(P<0.05)。结论超声联合微泡剂介导的猪自体BMSC心肌内移植有所增强,该方法为提高干细胞移植归巢效率提供一种新的思路。
Objective To investigate the effect of ultrasound microbubble contrast medium on intramyocardial transplantation of porcine bone marrow stem cells (BMSCs). Methods Eight Chinese 2-month-old domestic pigs were randomly divided into control group (group A, n = 4) and experimental group (group B, n = 4). Bone marrow was collected and isolated, purified, and cultured. BMSC and SPIO markers were obtained. Acute myocardial infarction model was established by interventional method. The BMSC (1MHz, 2W / cm2, 90s radiation) was transplanted into the B group under the action of ultrasound microbubbles. The AMS group was not subjected to BMSC transplantation by ultrasonic microbubbles. Animals in both groups were examined by magnetic resonance imaging (MRI) 24 h postoperatively, compared with BMSC transplantation. Animals were then sacrificed, myocardial infarction area, peripheral infarction area and normal area were examined by HE staining and Prussian blue staining. BMSCs were counted after blue staining. Results ①SPIO successfully labeled stem cells, therefore, living magnetic resonance imaging BMSC possible. ② Magnetic resonance imaging: In group A and group B, SPIO-labeled low density area was found in myocardial infarction area and peripheral infarction area, while low signal area was more in group B than in group A. ③ Prussian blue staining: two groups of myocardial digestion smear staining, BMSC in group B than in group A significantly increased (P <0.05). Conclusion Ultrasound combined with microbubble-mediated intraperitoneal transplantation of porcine autologous BMSC has been enhanced. This method provides a new idea for improving homing efficiency of stem cell transplantation.