The Study Of In Vitro Cytotoxicity Of Bur-like HAP Microsphere On The MC3T3-E1Cells

Hydroxyapatite materials have been widely used in the biomedical field, but the researchabout biocompatibility of HAP materials is not perfect. In this paper, Bur-like HAPMicrosphere (BHA) have been synthesized by a facile hydrothermal process, and the physicaland chemical properties of Bur-like HAP Microsphere (BHA) were characterized by SEM andSEM. In order to investigate the cytotoxicity of BHA, MC3T3-E1were incubated withdifferent concentration of BHA suspension for different time. Then, the MC3T3-E1cells weredetected with a series of methods such as3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test(MTT)、flow cytometry (FCM)、colocalization、AO/EB staining、single cell gel electrophoresis etc. The effects of BHA on the cell viability、 cellapoptosis/necrosis、DNA damage、level of reactive oxygen species (ROS)、mitochondrialmembrane potential and lysosome permeability were obtained:1. According to the results of SEM and Confocal and uptake experiments，it can beconfirmed that the particles existed inside the cell were debris of BHA. And these dibris wereendocytosed via clathrin and pinocytosis mediated endocytosis. These dibris mainly locate inmitochondria and lysosome. BHA can produce cytotoxicity by influencing the mitochondriaand lysosome membrane permeability of cells2. BHA shows cytotoxicity on the MC3T3-E1. After incubation with BHA particles，theviability of MC3T3-E1was significantly inhibited. BHA can also block the cell cycle in Sphase, promote cell apoptosis and necrosis, and damage DNA.3. The cytotoxicity of BHA is affected by its debirs. After treated with ultrosonic for30min，the content of debris was enhanced，and then the inhibition effect on cells and thelevel of ROS were also enhanced.