EC is involved in the immune and inflammatory response, coagulation, growth regulation, production of extracellular matrix components, and is a modulator of blood flow and blood vessel tone. EC injury, activation, or dysfunction is a hallmark of many pathologic

states including atherosclerosis, loss of semi-permeable membrane function, and thrombosis ACP-196 concentration [25]. A wide variety of stimuli can induce programmed cell death (apoptosis) of endothelial cells through extrinsic (death receptor) and/or intrinsic (mitochondria) apoptotic pathway, which is ultimately executed by the intracellular proteases called caspases. There also exist caspase-independent pathways 4SC-202 cell line of apoptosis and anti-apoptotic

proteins, which can protect cells from apoptosis. These pathways and proteins compose the complicated network of the cell apoptosis [26–29]. When injecting MNPs into blood vessels, ECs is the first tissue barrier encountered by the MNPs. The focus of this study is thus on the cytotoxicity evaluation of DMSA-coated Fe2O3 nanoparticles (DMSA-Fe2O3) on human aortic endothelial cell (HAEC), which is able to proliferate for many generations maintaining its endothelial characteristic and is widely used for in vitro study [30]. Methods Materials Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from GIBCO Company (Grand Island, New York, USA). Endothelial cell growth supplement (ECGS) was supplied by M&C Gene Technology (Beijing, China). MEM non-essential amino acid solution (100×), l-glutamine, thiazolyl blue tetrazolium bromide, haematoxylin, penicillin, and streptomycin were obtained from Sigma-Aldrich (St Louis, MO, USA). NVP-LDE225 cell line Prostacyclin I-2 (PGI-2), endothelin-1 (ET-1), and nitric oxide (NO) assay kits were obtained from Nanjing Jiancheng Bioengineering Institute Acyl CoA dehydrogenase (Nanjing, China). Primers were

synthesized by Sangon Biotechnology Co., Ltd. (Shanghai, China), and RNAiso Plus reagent, PrimeScript™ RT reagent Kit, and SYBR Premix Ex Taq™ were from TaKaRa Biotechnology Co., Ltd. (Dalian, China). Matrigel basement membrane matrix was from Becton Dickinson (Bedford, MA, USA). Preparation of DMSA-Fe2O3 nanoparticles The DMSA-Fe2O3 was prepared by co-authors Dr. Fei Xiong, Dr. Yu Zhang, and Dr. Ning Gu. The characterization data, such as transmission electronic microscopy (TEM) images, crystal structure, surface charge, and magnetic measurements and Fourier transform infrared spectroscopy measurements were determined as the previous report in Dr. Gu’s Lab [31].