氮掺杂硅量子点在血管内皮细胞成像中的应用

口腔生物医学 ›› 2022, Vol. 13 ›› Issue (2): 79-85.

氮掺杂硅量子点在血管内皮细胞成像中的应用

谷德奥¹,主璧君¹,禹怡君¹,苗雷英¹,²,刘超¹

1. 南京大学医学院附属口腔医院
2. 南京大学医学院附属口腔医院?南京市口腔医院牙体牙髓病科

收稿日期:2022-03-23 修回日期:2022-04-28 出版日期:2022-06-25 发布日期:2022-07-07
通讯作者: 刘超 E-mail:dxliuchao@163.com
基金资助:
国家自然科学基金;南京市卫生厅医学科技发展基金资助一般项目

Application of N-doped silicon quantum dot in vascular endothelial cells imaging

Received:2022-03-23 Revised:2022-04-28 Online:2022-06-25 Published:2022-07-07

摘要/Abstract

摘要： 目的：探究氮掺杂硅量子点（N-SiQD）作为人脐静脉内皮细胞（HUVEC）活细胞荧光标记材料使用的可行性及效能。方法：采用透射电子显微镜、荧光分光光度计对N-SiQD进行表征。体外培养HUVEC，与不同浓度N-SiQD共培养，通过CCK-8法及Calcein-AM/PI染色评估N-SiQD的生物安全性，同时观察其对HUVEC血管形成功能的影响，并对血管形成基因表达进行分析，通过共聚焦显微镜检测N-SiQD对HUVEC的体外荧光标记效应。结果：N-SiQD为平均直径4.62 nm的纳米球粒，分散度良好，在488 nm激发光照下呈绿色荧光。在共孵育24 h后，与对照组比较，浓度≤800μg/mL组的N-SiQD作用下细胞活性无影响，差异无统计学意义（P＞0.05），浓度≥1 000μg/mL时N-SiQD影响细胞增殖能力，差异有统计学意义（P＜0.05）；400 μg/mL N-SiQD对细胞血管形成功能有促进作用，VEGFA基因表达升高，VEGFB基因表达下降，差异具有统计学意义（P＜0.05）；400 μg/mL的N-SiQD可以很好地分布于细胞浆，发出稳定绿色荧光；在488 nm激发波长下，N-SiQD在30 min内可维持初始荧光强度的40%左右。结论：新合成N-SiQD具有良好的生物相容性，活细胞荧光标记效果良好，适宜的浓度下促进HUVEC的血管形成功能，可用于HUVEC活细胞标记。

关键词: 氮掺杂硅量子点, 脐静脉内皮细胞, 活细胞标记, 血管新生

Abstract: Objective:To explore the feasibility and efficacy of N-doped silicon quantum dots (N-SiQD) as fluorescent labeling materials for living cells of human umbilical vein endothelial cell (HUVEC). Methods: N-SiQD was characterized by transmission electron microscope and fluorescence spectrophotometer. HUVEC was cultured in vitro, and then co-cultured with different concentrations of N-SiQD. The CCK-8 and Calcein-AM/PI staining were used to evaluate the biosecurity of N-SiQD. Meanwhile, the effects of N-SiQD on tube formation of HUVEC were also observed, and the expression of angiogenesis genes was analyzed. Finally, the fluorescence labeling properties of N-SiQD on HUVEC in vitro was detected by confocal microscopy. Results: The N-SiQD as spherical nanoparticles with superior water-dispersibility was about 4.62 nm, and it showed green fluorescence under 488 nm exciting light. CCK-8 results showed that after 24 h co-incubation, compared with the control group, the activity of N-SiQD groups with a concentration of less than 800 μg/mL showed no statistical significance, while N-SiQD groups with a concentration of more than 1 000 μg/mL showed significant influence on cell proliferation (P<0.05). 400 μg/mL N-SiQD promoted the tube formation of HUVEC, and VEGFA gene expression was increased while VEGFB gene expression decreased, with statistical significance (P<0.05). Confocal microscopy showed that 400 μg/mL N-SiQD distributed well in the cytoplasm and emitted stable green fluorescence. At the excitation wavelength of 488 nm, N-SiQD could maintain about 40% of the initial fluorescence intensity within 30 min. Conclusions:The newly synthesized N-SiQD has good biocompatibility and fluorescence labeling function on living cells, and it may promote the behavioral function of HUVEC at appropriate concentration. Therefore, N-SiQD can be used to label living cells of HUVEC.

Key words: N-doped silicon quantum dot, umbilical vein endothelial cell, living cell labeling, angiogenesis

谷德奥主璧君禹怡君苗雷英刘超. 氮掺杂硅量子点在血管内皮细胞成像中的应用[J]. 口腔生物医学, 2022, 13(2): 79-85.

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