Ultrasound-mediated microbubble promotes bone marrow mesenchymal stem cell homing in the treatment of ischemic stroke

doi:10.3969/j.issn.2095-4344.2017.25.012

Abstract

Abstract:

BACKGROUND: In animal experiments, ultrasound-mediated microbubbles can promote the homing of transplanted stem cells to the ischemic area, enhance angiogenesis and small arterial formation, improve local blood flow in the ischemic myocardium and restore myocardial contractility.
OBJECTIVE: To investigate the effect of ultrasound-mediated microbubbles on intravenously transplanted bone marrow mesenchymal stem cell (BMSC) homing and the therapeutic efficiency on ischemic stroke.
METHODS: A middle cerebral artery occlusion (MCAO) model was induced by plug wire preparation. At 72 hours after MCAO, model rats were randomized into four groups: PBS group (n=15), BMSCs group (n=18), ultrasound+BMSCs group (n=18), ultrasound+microbubble+BMSCs group (n=18). Corresponding treatment was done in each group: 2 mL of PBS was injected via tail vein in the PBS group; about 3×10⁶ BMSCs diluted by 2 mL of PBS were injected via tail vein slowly in the BMSCs group; after skull ultrasound radiation (1 MHz, 2 W/cm²) for 120 seconds, BMSCs were injected via tail vein slowly in the ultrasound+BMSCs group; the same process as the ultrasound+BMSCs group was done following intravenous injection of 0.1 mL/kg microbubbles in the ultrasound+microbubble+BMSCs group.
RESULTS AND CONCLUSION: (1) Forty-eight hours after BMSCs transplantation, the BMSCs homing rate in the brain was significantly higher in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (2) Twenty-eight days after MCAO, nerve damage was significantly milder in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (3) Seven days after transplantation, the water content in the brain tissue was significantly lower in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (4) Seven days after transplantation, the cerebral infarction volume was significantly reduced in the ultrasound+microbubble+BMSCs group compared with the other two groups (P < 0.05). To conclude, ultrasound-mediated microbubbles can enhance the homing effect of intravenously transplanted BMSCs, reduce cerebral edema and cerebral infarction volume, improve the neurological function, and increase the therapeutic effect of BMSCs transplantation on ischemic stroke.

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

Key words: Brain Ischemia, Bone Marrow, Mesenchymal Stem Cell Transplantation, Microbubbles, Sonication, Tissue Engineering

CLC Number:

R394.2

Qian Jian, Fan Guo-feng, Xu Peng, Li Qi-ming, Sha Du-juan, Wang Jun, Zhang Jun. Ultrasound-mediated microbubble promotes bone marrow mesenchymal stem cell homing in the treatment of ischemic stroke[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(25): 4007-4012.

Figures/Tables 2

2.1 实验动物数量分析共实施造模SD大鼠76只。术中因完全切断颈总动脉失败2例，术后72 h以内因严重神经功能缺损死亡3只，继发颅内出血死亡2只，术后72 h共有69只大鼠存活，存活率为90.8%。存活72 h以上的大鼠进入分组实验，中途无脱落。 2.2 超声微泡干预增加骨髓间充质干细胞的归巢率荧光显微镜观察各组大鼠脑缺血区域冰冻切片显示(图1)，缺血区脑组织内出现散在绿色荧光颗粒为GFP标记的骨髓间充质干细胞。超声+微泡+骨髓间充质干细胞组积分吸光度(224 363.87±19 723.97)明显高于骨髓间充质干细胞组(199 016.21±16 918.42)、超声+骨髓间充质干细胞组 (205 330.94±13 957.45)，差异有显著性意义(P < 0.05)，骨髓间充质干细胞组和超声+骨髓间充质干细胞组比较差异无显著性意义。提示超声+微泡+骨髓间充质干细胞组归巢细胞明显增多。 2.3 超声微泡干预减轻大脑中动脉栓塞模型大鼠神经损害严重程度在造模后14 d，各组mNSS分值开始出现差异。骨髓间充质干细胞组、超声+骨髓间充质干细胞组在造模后14，21，28 d的神经损害程度均轻于PBS组，但仅28 d差异有显著性意义(P < 0.01)；超声+微泡+骨髓间充质干细胞组造模后14，21，28 d的神经损害程度均轻于PBS组，差异均有显著性意义(P < 0.01)；超声+微泡+骨髓间充质干细胞组造模后28 d的神经损害程度较骨髓间充质干细胞组、超声+骨髓间充质干细胞组更轻，差异均有显著性意义(P < 0.05)；骨髓间充质干细胞组、超声+骨髓间充质干细胞组比较差异均无显著性意义，见表1。 2.4 超声微泡干预降低大脑中动脉栓塞模型大鼠脑组织含水率各组大鼠脑组织含水率在造模后升高。在造模后 6 d即干细胞移植后3 d为著，并可见移植后7 d含水率降低。在移植后1，3 d，各组含水率差异无显著性意义(P > 0.05)；在移植后7 d，各组含水率均较PBS组低，与骨髓间充质干细胞组、超声+骨髓间充质干细胞组比较，超声+微泡+骨髓间充质干细胞组含水率显著降低，其差异有显著性意义(P < 0.05)；骨髓间充质干细胞组、超声+骨髓间充质干细胞组比较差异均无显著性意义，见表2。 2.5 超声微泡干预缩小大脑中动脉栓塞模型大鼠脑梗死体积细胞移植后7 d(造模后10 d)梗死累及范围主要为额顶叶的背外侧皮质、尾壳核、下丘脑部分，严重时累及到海马区(图2)。测量后发现超声+微泡+骨髓间充质干细胞组脑梗死体积为(15.40±0.88)%，与骨髓间充质干细胞组(17.00±0.63)%，超声+骨髓间充质干细胞组(17.31± 0.95)%，PBS组(19.51±1.10)%比较，梗死体积明显缩小，差异有显著性意义(均P < 0.05)，见表3。"

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