超声联合全氟戊烷相变纳米粒治疗冠状动脉血栓栓塞的溶栓效率及安全性研究
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武汉大学人民医院

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国家自然科学基金青年基金项目(No. 81501495);湖北省卫生计生委联合基金面上项目(No.WJ2018H0078)


Thrombolytic efficiency and biosafety of ultrasound combined with dodecafluoropentane phase-change nanoparticles in the treatment of coronary embolism: an in-vitro experimental study
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renmin hospital of wuhan university

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National NatureScienceFoundationofChina (No. 81501495);

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    摘要:

    目的 探索超声联合相变纳米粒靶向助溶人工血管内血栓的溶栓效率,评价其对于急性心肌梗死(AMI)冠状动脉血栓栓塞治疗的有效性及生物安全性。方法 取家兔颈总动脉血制备成统一规格约0.2ml条形血栓块;采用声振乳化法制备全氟戊烷(DDFP)脂质相变型纳米粒;实验分为三组:PBS对照组(A组),声诺维微泡组(B组),DDFP相变纳米粒组(C组);将各组血栓条块置入体外溶栓模拟循环装置的人工血管中,同时联合低强度聚焦超声仪(LIFU)靶向辐照。优化LIFU各项参数(时间、功率等),综合LIFU和纳米粒的安全性探索最佳溶栓条件。采用Malvern激光测量仪等检测相变纳米粒粒径等一般性能;称量溶栓前后血栓重量评价溶栓效率并观察比较血栓HE染色内部结构;CCK-8评价不同浓度纳米粒和不同强度LIFU对脐静脉内皮细胞(HUVECs)存活率的影响。结果 相变纳米粒粒径大小为(542.6± 28.4)nm;荧光显微镜示纳米粒液体相时结构稳定,大小均一;当LIFU的参数调整为辐照时间:20min、功率为6W时,纳米粒的相变转化率、溶栓效率及HUVECs细胞的存活率均较高;溶栓后C组血栓HE染色显示红细胞及血小板梁数量显著减少,质地松软,结构疏松。结论 LIFU联合DDFP相变纳米粒可有效助溶冠状动脉内血栓,这种治疗方式不仅高效,且具有较好的生物安全性,为AMI提供了一种新的治疗方式。

    Abstract:

    Objective To explore the efficiency of ultrasound combined with dodecafluoropentane (DDFP) phase-change nanoparticles in the thrombolytic therapy, and to evaluate the effectiveness and the biological safety of the treatment in acute myocardial infarction (AMI). Methods Carotid artery blood of the rabbits were prepared into pieces of 0.2ml thrombus. The DDFP nanoparticles were prepared by acoustic emulsion method. There were three groups in the experiment: poly butylene succinate buffer (PBS) as blank control (Group A), SonoVue microbubbles as Group B and DDFP phase-change nanoparticles as Group C. Thrombus in all three groups were placed into an artificial blood vessel model in an extracorporeal thrombolysis simulator combined with low intensity focusing ultrasonic instrument (LIFU) irradiation. Different parameters (time, power, etc.) of LIFU were optimized regarding to the biosafety with LIFU and nanoparticles. The size of the nanoparticles was detected by Malvern laser granulator. The weights and pathological presentations were compared before and after thrombolysis. Cell counting kit-8 (CCK-8) were used to evaluate the survival rates of human umbilical vein endothelial cells (HUVECs) with different concentrations of nanoparticles and different parameters of LIFU. Results The average size of DDFP nanoparticles was(542.6± 28.4)nm. Fluorescence microscopy showed that the structure of nanoparticles was stable and uniformed in size. When the parameters of LIFU were adjusted to 20min in irradiation time and 6W in power, the phase transformation rates, thrombolysis efficiency and the survival rates of HUVECs were higher than those of other conditions. HE-staining slides showed that the number of red blood cells and platelets decreased significantly after thrombolysis in group C, also the texture and structure were softer and looser. Conclusion LIFU combined with DDFP phase-change nanoparticles can effectively dissolve coronary thrombosis. This novel treatment is not only efficient, but also better for biosafety, which has the potential to provide a new treatment for acute myocardial infarction.

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姜楠,胡波.超声联合全氟戊烷相变纳米粒治疗冠状动脉血栓栓塞的溶栓效率及安全性研究[J].临床超声医学杂志,2019,21(5):321-330

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  • 收稿日期:2019-01-16
  • 最后修改日期:2019-01-16
  • 录用日期:2019-03-07
  • 在线发布日期: 2019-05-22
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