Genotoxicity of organic bentonite particles in vitro / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the genotoxicity induced by organic bentonite particles in vitro.</p><p><b>METHODS</b>Human B lymphoblast cells (HMy2.CIR) were exposed to organic bentonite particles at the doses of 0, 1.88, 3.75, 7.50 and 15.00 µg/ml for 24, 48 and 72 h, calcium sulfate (30 µg/ml) and SiO2 (30 and 240 µg/ml) served as negative and positive controls, respectively. The genotoxicity of organic bentonite particles and soluble fraction was detected using comet assay and Cytokinesis-block micronucleus (CBMN) assay.</p><p><b>RESULTS</b>The results of comet assay indicated that % tail DNA increased with the exposure doses and time in organic bentonite group, % tail DNA at the dose of 15.00 µg/ml for 24 h, 48 h and 72 h in organic bentonite group were 3.20 ± 0.19, 4.63 ± 0.88 and 9.49 ± 1.31 respectively which were significantly higher than those in calcium sulfate group (1.40 ± 0.11, 1.37 ± 0.22 and 0.90 ± 0.16) and those in 30 µg/ml SiO2 group (1.83 ± 0.21, 1.41 ± 0.27 and 2.48 ± 0.25) (P < 0.01). The results of CBMN assay showed that micronucleus frequencies (MNF) in organic bentonite group (except for 1.88 µg/ml for 24 h) were significantly higher than those in 30 µg/ml calcium sulfate group (MNF for 24, 48 and 72 h were 1.33‰ ± 0.58‰, 1.33‰ ± 1.15‰ and 1.33‰ ± 0.58‰) and those in 30 µg/ml SiO2 group (2.00‰ ± 0.00‰, 1.68‰ ± 0.58‰ and 2.33‰ ± 0.58‰) (P < 0.01). The results of two assays demonstrated that the soluble fraction of organic bentonite did not induce the genotoxicity.</p><p><b>CONCLUSION</b>The organic bentonite dusts can induce the genotoxicity in vitro, which may be from the particle fraction.</p>

Cyto-genotoxicity induced by cigarette smoke condensates in human peripheral blood lymphocytes in vitro / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the cyto-genotoxicity of cigarette smoke condensates (CSCs) in human peripheral blood lymphocytes with different assays in vitro.</p><p><b>METHODS</b>Human lymphocytes were exposed to particle matter of cigarette smoke combined with or without S9 mixtures at doses of 25, 50, 75, 100 and 125 microg/ml for 3 h. The cytotoxicity induced by CSCs was detected by CCK-8 assay. The DNA damage, DNA repair (repair time: 30, 60, 90, 120 and 240 min, respectively) and the somatic cell mutations induced by 75 microg/ml CSCs were measured by comet assay, hprt gene and TCR gene mutation tests, respectively.</p><p><b>RESULTS</b>CCK-8 assay indicated that the cell viability decreased with CSCs doses. At the doses of 100, 125 microg/ml, the cell viability of CSCs +S9 group was significantly higher than that of CSCs -S9 group (P < 0.05, P < 0.01). In comet assay, DNA damage significantly increased in a dose-dependent manner, as compared with controls (P < 0.01). Moreover, there was significant difference between -S9 group and +S9 group (P < 0.05, P < 0.01). The Mf-TCR at each dose group was significantly higher than that of controls (P < 0.05, P < 0.01). The Mf-hprt at high-dose groups were significantly higher than that of controls (P < 0.01), and significant difference of Mf-TCR and Mf-hprt at high doses of CSCs between -S9 group and +S9 group (P < 0.05, P < 0.01). The DNA damage induced by CSCs +S9 or CSCs -S9 could be repaired, but DNA repair speed was different between -S9 group and +S9 group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>CSCs may induce cyto-genotoxicity in human peripheral blood lymphocytes in vitro, but S9 mix could reduce the toxicity of CSCs and impact DNA repair speed.</p>

Comparative study of cytotoxicity induced by two kinds of bentonite particles in vitro / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study comparatively the cytotoxicity induced by acid bentonite and organic bentonite.</p><p><b>METHODS</b>The cytotoxicity of two kinds of bentonite was detected using CCK8 assay, neutral red uptake (NRU) assay, lactate dehydrogenase (LDH) leakage assay, apoptosis assay and hemolysis assay. In hemolysis assay human erythrocytes served as target cells and were exposed to the two kinds of bentonite at the doses of 0, 0.3125, 0.6250, 1.2500 and 2.5000 mg/ml for ten min. In other four assays, human B lymphoblast cells (HMy2.CIR) served as target cells and were exposed to the two kinds of bentonite at the doses of 0, 10, 20, 30, 60, 120 and 180 microg/ml for four h.</p><p><b>RESULTS</b>In hemolysis assay, the hemolysis rates induced by two kinds of bentonite at all doses were significantly higher than that of control (P<0.05); in CCK-8 assay, the cellular activities in acid bentonite group at the doses > or =30 microg/ml and in organic bentonite group at the doses > or =20 microg/ml were significantly lower than that of control (P<0.01); the similar results appeared in NRU assay and LDH assay, and the dose-effect relationship was observed in above 4 assays. In apoptosis assay, the early apoptosis cell rates in acid bentonite group at the dose of 180 microg/ml and in organic bentonite group at the doses of 120,180 microg/ml were significantly higher than that of control (P<0.05). Moreover, the results of five in vitro assays indicated the cytotoxicity induced by organic bentonite was higher than that induced by acid bentonite.</p><p><b>CONCLUSION</b>Two kinds of bentonite could induce cytotoxicity, such as apoptosis and damage of cell membrane. The cytotoxicity of organic bentonite is higher than that of acid bentonite due to the different industrial treatment and characteristics of two kinds of bentonite particles.</p>