Study on Biological Safety of Functionalized Nano-Hydroxyapatite
Zhao Yanzhong1, 2* Yang Min1 Wang Guohui1 Tan Juan1 Zhu Shaihong1, 2
1Medical Experimental Center of theThird Xiangya Hospital, Central South University, Changsha 410013, China 2Research Center for Medical Material and Instruments, Central South University, Changsha 410013, China
Abstract To investigate the feasibility of nano-hydroxyapatite with arginine-functionalized and Tb3+doped HAP/Arg as a gene drug carrier, and to study the biological safety of HAP/Arg. Normal human vascular endothelial cells (HAEC) and human tumor cells (Hela) were selected as cell models in vitro, under the exposure of 0, 25, 50 to 200 μg/mL of HAP/Arg nanoparticles, the cell survival rate was measured by the enzyme labeled immunoassay instrument, and the structural integrity of the cell membrane of which was examined by LDH method. Healthy mice were divided into low dose group (10 μg/mL/20 g weight), middle dose group (100 μg/mL/20 g weight), high dose group (500 μg/mL/20 g weight ) and control group, to count number of deaths, to detect serum biochemical indexes and to observe pathologic features of main organs by inverted fluorescence microscope. According to the results of acute toxicity experiment, healthy mice were divided into low dose group (0.09 mg/(kg·d)), middle dose group (4.5 mg/(kg·d)), high dose group (225 mg/(kg·d) ) and control group, according to the preclinical safety evaluation standard to judge its reproductive toxicity. The results showed that HAP/Arg with concentration of 0, 25, 50 and 200 μg/mL did not affect normal growth and cell membrane structure under 4, 24, 48 and 72 h of HAEC cells and Hela cells. In experiments on animal acute toxicity and general reproductive toxicity, there were no obvious toxic reaction and death in the mice of each dose group and control group, the inflammation and injury were not found in the main organs. Statistical analysis shows that each dosage groups of HAP/Arg has no significant difference compared with control group (P>0.05). Above results are expected to be useful for developing a novel efficient, safe and convenient gene delivery system for the next research step.
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