6533b851fe1ef96bd12a946b

RESEARCH PRODUCT

Cell imaging connected with protein mapping to investigate glyphosate induced toxicity on human keratinocytes

subject

description

During the skin ageing which is accelerated by environmental factors, the generated reactive oxygen species make unstable the "proliferation/differentiation/apoptosis" coupling of epidermal cells. A biochemical study, previously realized on human keratinocyte cultures treated by a pesticide, glyphosate, allowed the validation of an in vitro model of skin ageing. In this work, we propose to study the loss of HaCaT cell integrity appearing after this chemical induced-oxidative stress. An original approach, combining a micro- to nanoscale cell characterization and analysis of fluctuating protein expression, is proposed: a) the exploration of cell growth and morphology through fluorescence and AFM imaging, b) a functional investigation using force spectroscopy for detection of oxidative stress biomarkers in membranes and c) the establishment of proteomic comparative profiles between safe and glyphosate-treated cells. Our results revealed up to now that a glyphosate treatment induced cell impairment in an acute dose-dependency. Indeed, we observed a breakage of plasma membrane and a release of cytosol, microtubules disorganization and fragmentation, and finally aggregation of chromatin. These observations characterized apoptotic key events. Our first force spectroscopy results revealed that a glyphosate coated tip interacted specifically with keratinocyte membrane structures. This nanobiotechnological tool will allow cell mapping establishment and screening original anti-oxidant molecules. The proteomic profiling would highlight cell oxidative stress cascade and identify corresponding major biomarkers in our skin ageing model. Original molecules with cytoprotective potentials will be tested and compared to well-known antioxidants, in order to correct pesticide induced cutaneous disorders. These data all together will contribute to better encircle the mechanisms of chemical and environmental hazards acting on the human skin.