Toxicological Sequelae of Pesticide Combinations Exposure in Buffalo Mesenchymal Stem Cells under In Vitro

The presence of one or more pesticides in a variety of mediums is responsible for their indirect toxicological events leading to cell senescence. In the present investigation, the endeavor was made to see the effect of pesticides Car-Benda-Zim (CBZ) and IMIdacloprid (IMI) alone and in combination with bone marrow-derived Mesenchymal Stem Cells (bMSCs) of buffalo origin. Isolated and cultured bMSCs were exposed to CBZ and IMI alone and in combinations at lower doses. Cells were observed for alterations in cell morphology, oxidative stress, mitochondrial damage and cellular senescence. bMSCs characterized for stem cell surface markers and found to be positive for AP, CD73 and OCT4. bMSCs exposed to IC25, IC12.5 and IC6.25 CBZ and IMI alone and combinations of IC12.5 and IC6.25 of CBZ and IMI. Results revealed significant reduction (p≤0.05) in cell viability noticed on microscopic examination along with loss of normal cell morphology and increased in Reactive Oxygen Species (ROS) positive cells, cells with loss of ΔΨm and number of senescent cells in CBZ and IMI treated groups. Lower dose combination groups showed elevated effects when compared with higher dose alone treated groups and control groups. Present findings suggest that CBZ and IMI induced cytotoxicity in bMSCs mediated via ROS production, altered ΔΨm leading to the cell damage and predisposing senescence process. Moreover, the co-existence of CBZ and IMI in a medium has a considerably more toxic effect than their individual effect.