Yang Min, Zhao Yanzhong, Wang Guohui, Tan Juan, Zhu Shaihong
Medical Experimental Center, Third Xiangya Hospital, Central South University, Changsha 410013, China.
Medical Experimental Center, Third Xiangya Hospital; Research Center for Medical Material and Instruments, Central South University, Changsha 410013, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2016 Sep 28;41(9):937-45. doi: 10.11817/j.issn.1672-7347.2016.09.008.
To explore the interaction between arginine functionalized hydroxyapatite (HAP/Arg) nanoparticles and endothelial cells, and to investigate mechanisms for endocytosis kinetics and endocytosis.
Human umbilical vein endothelial cells (HUVECs) were selected as the research model.Cellular uptake of HAP/Arg nanoparticles were observed by laser scanning confocal microscopy.Average fluorescence intensity of cells after ingestion with different concentrations of HAP/Arg nanoparticles were determined by flow cytometer and atomic force microscopy.
The HAP/Arg nanoparticles with doped terbium existed in cytoplasm, and most of them distributed around the nucleus area after cellular uptake by HUVECs. Cellular uptake process of HAP/Arg nanoparticles in HUVECs was in a time and concentration dependent manner. 4 h and 50 mg/L was the best condition for uptake. HAP/Arg nanoparticles were easier to be up-taken into the cells than HAP nanoparticles without arginine functionalized.
HAP/Arg nanoparticles are internalized by HUVECs cells through an active transport and energy-dependent endocytosis process, and it is up-taken by cells mainly through caveolin-mediated endocytosis, but the clathrin-dependent endocytic pathway is also involved..
探讨精氨酸功能化羟基磷灰石(HAP/Arg)纳米颗粒与内皮细胞之间的相互作用,并研究其胞吞动力学和胞吞作用机制。
选择人脐静脉内皮细胞(HUVECs)作为研究模型。通过激光扫描共聚焦显微镜观察HAP/Arg纳米颗粒的细胞摄取情况。用流式细胞仪和原子力显微镜测定不同浓度HAP/Arg纳米颗粒摄取后细胞的平均荧光强度。
掺杂铽的HAP/Arg纳米颗粒存在于细胞质中,HUVECs摄取后大部分分布在细胞核区域周围。HUVECs中HAP/Arg纳米颗粒的细胞摄取过程呈时间和浓度依赖性。4小时和50毫克/升是摄取的最佳条件。与未进行精氨酸功能化的HAP纳米颗粒相比,HAP/Arg纳米颗粒更容易被细胞摄取。
HAP/Arg纳米颗粒通过主动转运和能量依赖的胞吞过程被HUVECs细胞内化,其主要通过小窝蛋白介导的胞吞作用被细胞摄取,但网格蛋白依赖性内吞途径也参与其中。