人血清白蛋白用于动脉斑块内骨桥蛋白分子成像的实验研究

Application of HSA-based nanoparticles for detecting atherosclerotic plaques

目的通过构建基于人血清白蛋白(HSA)的光学纳米探针(ICG-HSA-OPN),靶向合成型平滑肌细胞,实现对动脉易损斑块的精准识别。方法 HSA耦联骨桥蛋白(OPN)活性短肽后,于水溶液中通过静电作用包载吲哚菁绿(ICG)自组装为纳米颗粒。用透射电镜和紫外光谱对探针进行表征,CCK-8实验检测探针对MOVAS细胞系的毒性,激光共聚焦显微镜观察细胞摄取能力。结果纳米颗粒呈球形,粒径均一,大小约为20 nm;紫外光谱显示ICG已成功包载入HSA纳米探针,CCK-8结果表明不同浓度纳米探针(ICG 0～100μg/ml)的细胞存活率与对照组的差异无统计学意义(P≥0.05);激光共聚焦显微镜下可见靶向OPN纳米探针组细胞摄取能力明显强于非靶向组。结论本研究成功构建了能够特异性靶向易损斑块内合成型平滑肌细胞的纳米探针,且生物相容好。

Objective To detect vulnerable plaques by constructing synthetic smooth muscle cells targeted HSA-based nanoparticles(NPs). Methods ICG-HSA-OPN NPs were prepared by targeting OPN peptide,encapsulating fluorescent dye (ICG) and self-assembling into nanoparticles in PBS solution. Characterization was measured by the transmission electron microscopy (TEM) and ultraviolet(UV) absorption spectrum. Biocompatibility was evaluated in MOVAS cell line by CCK-8 essay. Specificity and cell uptake capacity were observed by laser scanning confocal microscope in vitro. Results The TEM image and UV spectrum manifested the discoid-shaped NPs were well-preparedandabout 20nm in size. CCK-8 assay showed there was no significant difference between OPN targeted group and control group in cell viability(P ≥ 0.05). Good binding specificity were demonstrated according to cellular uptake analysis. Conclusion We have successfully constructed HSA-based nanoparticles for fluorescence imaging of synthetic smooth muscle cells in vulnerable plaques which possessing excellent biocompatibility.