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Interestingly, the Fe2O3 and silicon dioxide NPs have a similar significant decrease in cell viability and the lowest reduction in cellular viability was observed with Al2O3 NPs. The Co3O4 NPs were found to be more toxic to human lymphocytes followed by Fe2O3, silicon dioxide, and Al2O3 NPs in a descending order. The cytotoxicity was observed in human lymphocytes after 24 h of interaction of NPs by MTT and LDH assays. Both MTT and LDH assays have been frequently used for determining the in vitro cytotoxicity of NPs in cell culture experiments. Results of MTT assay showed a significant reduction in cell viability compared with control in a dose-dependent manner. At 50, 75, and 100 mg/ml, the Co3O4 NPs showed asignificant reduction in percentage cell viability (p <0.001).

The Co3O4 NPs showed a significant increase (p < 0.001) in LDH level when compared with Fe2O3 NPs and silicon dioxide NPs after 24 h of exposure to the human lymphocytes at 50, 75, and 100 mg/ml. The cytotoxicity was further investigated by estimating the percentage of LDH released after incubation with all NPs. The Al2O3 NPs showed a significant increase in LDH level (p < 0.01) when exposed to 100 mg/ml,whereas an insignificant increase in LDH level (p > 0.05) was observed when treated at 50 and 75 mg/ml in human lymphocytes. Cellular viability in human lymphocytes by MTT assay when treated with Co3O4, Fe2O3, silicon dioxide(SiO2) NPs, and Al2O3 NPs. The data obtained are expressed in terms of mean SD (n = 3). Each were test three times.

The Fe2O3 NPs and silicon dioxide(SiO2) NPs showed a significant increase in ROS level at 75 and 100 mg/ml (p <0.001), but insignificant increase in ROS level was observed at 50 mg/ml (p > 0.05). However the Co3O4 NPs were found to significantly increase (p < 0.001) ROS level in the human lymphocytes at 50, 75, and 100 mg/ml. The Al2O3 NPs showed a significant increase in ROS level at 100 mg/ml (p < 0.01). These are the oxidative stress in human lymphocytes.



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