以二氧化矽為模板合成具標靶性微米氣泡於超音波之應用

Preparation of multi-targeted ultrasound contrast agent using silica particles as template

陳琬琪(Wan-Chi Chen)

指導教授 ： 林政鞍 ； 張恒雄

中原大學/工學院/生物醫學工程研究所/碩士(2015年)

https://doi.org/10.6840/cycu201500940

摘要

肝癌是常見的癌症，目前最普遍的診斷方式是超音波檢測，但脂肪肝會影響超音波傳導，所以超音波只能檢查到較表淺處有無腫瘤。而本研究即是以模板法調控對比劑粒徑。改善傳統超音波對比劑粒徑分佈不均，易造成血管阻塞之現象。先製備粒徑800 nm二氧化矽奈米模板，並在其表面包覆高分子，形成核殼層結構，接著以適當溶劑氫氟酸蝕刻模板，經由透析和冷凍乾燥後，即可獲得粒徑分佈較均一的氣泡結構。本研究發現在醫用超音波連續刺激下，該微氣泡在活體外造影成像時間可達15分鐘。並可由細胞毒性測試MTT證實該微氣泡具生物相容性。同時在合成微氣泡的過程中，以掃描式電子顯微鏡與穿透式電子顯微鏡觀察粒子的表面形態變化和驗證其中空結構。此外，亦於該微氣泡表面修飾有VEGFR2抗體，使其成為具有靶向性的微氣泡功能，能輸送到腫瘤血管進行標靶腫瘤，能使超音波影像增強對比度。而本研究為了確定Hep G2及HuH-7人類肝癌細胞具有VEGFR2 receptor，進行了VEGFR2蛋白的驗證，將靶向性微氣泡與Hep G2及HuH-7人類肝癌細胞共培養後，進行螢光顯微鏡拍攝驗證Anti-VEGFR2-MB在癌細胞上會有螢光表現。最後，將該靶向性微氣泡進行動物實驗，驗證具靶向性的微氣泡，在動物體內超音波影像能增強影像效果。本研究期望此超音波對比劑能讓肝腫瘤在超音波影像上能夠更為清晰，進而讓超音波影像的診斷更為準確。

關鍵字

對比劑 ； 微米氣泡 ； 靶向性 ； 超音波 ； 肝癌細胞 ； VEGFR2

並列摘要

Liver cancer is a common cancer, the most common diagnostic modality is ultrasound testing, but will affect the fatty liver ultrasound conduction, so ultrasound examination to more superficial only at whether the tumors. The present study is based on a template method that is the regulation of the contrast agent particle size. Traditional ultrasound contrast agent to improve the particle size distribution is uneven, easily lead to the phenomenon of vascular obstruction. Particle size of 800 nm is first prepared silica nano template, and a polymer coating on its surface, forming a core shell structure, in an appropriate solvent followed by hydrofluoric acid etching template, by dialysis and freeze-drying, the particle size can be obtained distribution is more homogeneous bubble structure. The study found that continuous stimulation in medical ultrasound, the microbubbles up to in vitro contrast imaging time 15 minutes. And by the MTT cytotoxicity test confirmed that the microbubbles with biocompatibility. At the same time during the synthesis of microbubbles in a scanning electron microscope and a transmission electron microscope to observe the surface morphology of particles of the hollow structure and validated. In addition, also in the microbubble surface modification have VEGFR2 antibody, making it the microbubbles have targeted function, can be delivered to the tumor target tumor blood vessels, enabling ultrasound image contrast enhancement. And this study to determine the Hep G2 and HuH-7 human hematoma cells VEGFR2 receptor, VEGFR2 protein was verified, after the targeted microbubbles and Hep G2 and HuH-7 human hematoma cells cultured, were shot fluorescence microscope Verify Anti-VEGFR2-MB will be reflected in the cancer cell fluorescence. Finally, the targeted microbubbles animal experiments, verify with targeted microbubbles, ultrasound imaging in vivo can enhance the image effect. This study was expected to make this ultrasound contrast agent in ultrasound imaging of liver tumors can be more clear, thereby allowing more accurate diagnostic ultrasound imaging.

並列關鍵字

contrast agents ； microbubble ； targeted ； ultrasound ； Liver cancer cell ； VEGFR2

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