Diethyl citrate and sodium citrate reduce the cytotoxic effects of nanosized hydroxyapatite crystals on mouse vascular smooth muscle cells.

摘要: Objective This study aimed to investigate the damage mechanism of nanosized hydroxyapatite (nano-HAp) on mouse aortic smooth muscle cells (MOVASs) and injury-inhibiting effects diethyl citrate (Et2Cit) sodium (Na3Cit) develop new drugs that can simultaneously induce anticoagulation inhibit vascular calcification. Methods The change in cell viability was evaluated using a proliferation assay kit, amount lactate dehydrogenase (LDH) released measured an LDH kit. Intracellular reactive oxygen species (ROS) mitochondrial were detected by DCFH-DA staining JC-1 staining. Cell apoptosis necrosis Annexin V calcium concentration lysosomal integrity Fluo-4/AM acridine orange, respectively. Results Nano-HAp decreased damaged membrane, resulting release large LDH. entered mitochondria, then induced producing ROS. In addition, nano-HAp increased intracellular Ca2+ concentration, leading rupture necrosis. On addition anticoagulant Et2Cit or Na3Cit, membrane potential increased, whereas released, ROS, rate decreased. Et2 Cit Na3Cit could also chelate with Ca+ elevations nano-HAp, prevent rupture, reduce High concentrations exhibited strong inhibitory effects. capacity stronger than at similar concentrations. Conclusion Both significantly reduced cytotoxicity MOVASs inhibited crystals. chelating function resulted both binding HAp. may play role as anticoagulants reducing injury wall caused nano-HAp.