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1. 恩施土家族苗族自治州中心医院泌尿外科
2. 恩施土家族苗族自治州中心医院放射科
纸质出版日期:2020,
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[1]程武松,陈洪波,游玉峰.载10-羟基喜树碱液态氟碳靶向纳米粒的制备及体外双模态成像[J].临床超声医学杂志,2020,22(09):641-646.
CHENG Wusong, CHEN Hongbo, YOU Yufeng. Preparation and in vitro dual-mode imaging of 10-HCPT-loaded liquid fluorocarbon nanoparticles[J]. Journal of Clinical Ultrasound in Medicine, 2020,22(9):641-646.
[1]程武松,陈洪波,游玉峰.载10-羟基喜树碱液态氟碳靶向纳米粒的制备及体外双模态成像[J].临床超声医学杂志,2020,22(09):641-646. DOI: 10.16245/j.cnki.issn1008-6978.2020.09.001.
CHENG Wusong, CHEN Hongbo, YOU Yufeng. Preparation and in vitro dual-mode imaging of 10-HCPT-loaded liquid fluorocarbon nanoparticles[J]. Journal of Clinical Ultrasound in Medicine, 2020,22(9):641-646. DOI: 10.16245/j.cnki.issn1008-6978.2020.09.001.
目的制备载10-羟基喜树碱(10-HCPT)液态氟碳靶向纳米粒
观察其对人卵巢癌SKOV-3细胞的寻靶能力及体外超声/CT双模态成像效果。方法以羟基端乳酸/羟基乙酸共聚物(PLGA-COOH)、全氟溴辛烷(PFOB)及10-HCPT为原料
采用双乳化法制备PLGA@10-HCPT-PFOB纳米粒
观察其外部形态和内部结构
测量粒径大小和表面电位
以及10-HCPT的包封率和载药量;通过碳二亚胺法连接cRGD肽制备cRGD-PLGA@10-HCPT-PFOB靶向纳米粒
使用共聚焦显微镜和流式细胞仪检测PLGA@10-HCPT-PFOB纳米粒与cRGD肽的连接情况。体外实验检测其对人卵巢癌SKOV-3细胞的靶向性能;观察靶向纳米粒的体外超声/CT成像能力。结果本实验制得的靶向载药纳米粒外观呈黄色混悬液
光镜下纳米粒形态规则、大小均一
平均粒径(322.03±5.34)nm
平均表面电位(-1.55±0.10)mV;扫描电镜示靶向纳米粒呈球形
表面光滑;透射电镜示其为核壳结构
PLGA包裹PFOB和10-HCPT
10-HCPT包封率和载药量分别为(81.34±2.28)%和(13.24±1.24)%;共聚焦显微镜观察示FITC标记的cRGD肽呈绿色荧光
与DiI标记染色的红色纳米粒共同融合呈橙黄色荧光
流式细胞仪检测PLGA@10-HCPT-PFOB纳米粒FITC荧光强度为0.78%
cRGD-PLGA@10-HCPT-PFOB纳米粒FITC荧光强度为80.76%;共聚焦显微镜及流式细胞仪显示大量纳米粒靶向到SKOV-3细胞表面
与细胞膜上的αvβ3受体结合
部分被SKOV-3细胞吞噬;随着靶向纳米粒的浓度不断增加
其体外超声/CT成像明显增强。结论本实验成功制备载10-HCPT液态氟碳靶向纳米粒
其10-HCPT包封率和载药量均较高
对人卵巢癌SKOV-3细胞有较强的靶向性
通过聚集显影可增强体外超声/CT成像。
Objective To prepare 10-hydroxycamptothecin(10-HCPT) loaded liquid fluorocarbon targetingnanoparticles
and to investigate their targeting ability to ovarian carcinoma cells(SKOV-3) and the effect of ultrasound/computed tomography(CT) dual-mode imaging in vitro.Methods By using hydroxy terminated lactic acid/glycolic acidcopolymer(PLGA-COOH)
perflurooctyl bromide(PFOB)and 10-HCPT
PLGA@10-HCPT-PFOB nanoparticles were preparedby double emulsion solvent evaporation method.The surface morphology and internal structure were observed
the particle size
surface potential
encapsulation efficiency and drug loading of 10-HCPT were measured as well. The cRGD peptide wasconjugated to the nanoparticles by carbodiimide technique. The binding of cRGD-PLGA@10-HCPT-PFOB was detected bylaser scanning confocal microscope and flow cytometer.The targeting properties of the nanoparticles to SKOV-3 were detected.The ability of ultrasound/CT imaging of targeted nanoparticles was evaluated in vitro.Results The targeted nanoparticles wereproduced as a yellow suspension.The size distribution of the nanoparticles was uniform observed by optical microscope
with amean diameter of(322.03±5.34)nm and surface potential was(-1.55±0.10)mV.The surface of the targeted nanoparticles wassmooth observed by the scanning electron microscope. The core-shell structure was observed by the transmission electron microscope
and the PFOB and 10-HCPT were trapped in PLGA.The drug encapsulation and drug loading capacity of 10-HCPTwere(81.34 ± 2.28)% and(13.24 ± 1.24)%
respectively. Laser scanning confocal microscope showed that the FITC-labeledcRGD peptides were green and it fused to orange with the red nanoparticles labeled DiI.The flow cytometer showed that theintensity of FITC in PLGA@10-HCPT-PFOB and cRGD-PLGA@10-HCPT-PFOB nanoparticles were 0.78% and 80.76%
respectively. The laser scanning confocal microscope and flow cytometry showed that a large number of nanoparticles weretargeted to SKOV-3 cells
binding to αvβ3 receptor
some of which were uptaken by SKOV-3 cells. With the increasingconcentration of targeted nanoparticles
the ultrasound and CT imaging were enhanced significantly in vitro.Conclusion The10-HCPT liquid fluorocarbon targeting nanoparticles are successfully prepared
and the encapsulation and loading capacityof 10-HCPT are both high. The nanoparticles have strongly specific affinity on the SKOV-3 cells and owned enhancedultrasound/CT imaging in vitro.
10-羟基喜树碱靶向纳米粒成像体外
10-hydroxycamptothecinTargetingNanoparticlesImagingIn vitro
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