Study of the effects of xenobiotics on attached mammalian cell line by label-free biosensor

Cytotoxicity measurements predominantly apply label-based techniques (for example enzymatic and MTT assays) and animal feeding tests. In this study, we attempted to develop a new and quick cytotoxicity determination method... [ view full abstract ]

Cytotoxicity measurements predominantly apply label-based techniques (for example enzymatic and MTT assays) and animal feeding tests. In this study, we attempted to develop a new and quick cytotoxicity determination method with Epic® BenchTop label-free optical biosensor, which eliminates the drawbacks of the conventional methods (Fig. 1.).[1] Digital laser holographic transmission microscopy was applied as a label-free, non-invasive, non-destructive and non-phototoxic method for qualitative and quantitative measurement of cell morphological changes.[2]

Environmental and toxicological characteristics of formulated pesticides may substantially differ from those of their active ingredients or other components alone. In the present work, Roundup herbicide, its formulant (polyethoxylated tallow amine, POEA) and its active ingredient (glyphosate) were studied on a mammalian cell lines.

Conventional method (MTT test) and holographic microscopy have proved the cytotoxicity effects of the agents on the neuroectodermal cell line.[3] Therefore, the cytotoxicity effect of this herbicide was intended to detect on the preosteoblastic cells by Epic Bt biosensor. During the Epic biosensor measurement, the reagents were added to the previously adhered on the biosensor surface cells. To characterize kinetic of cell detachment from the sensor surface a widely used for this purpose trypsin/EDTA solution was applied. Similar kinetics of biosensor signals were obtained for the treatment with xenobiotics and their magnitude and slope correlate well with the increased detachment of cells from the sensor surface. These results compare with those obtained with Holographic microscopy (Holomonitor). Holographic microscopy revealed that glyphosate influences cell division, motility, elongated morphology and causes slower cell death. Glyphosate has to be applied at higher concentration to result in cytotoxic effects than Roundup or POEA. In conclusion, the formulant POEA was determined more toxic than herbicide preparation Roundup or its active ingredient glyphosate.

[1] N. Orgovan et. al., Sci. Rep., 2014, 4: 4034.

[2] B. Peter et. al., J. Biomed. Opt., 2015, 20, 067002-067002.

[3] I. Székács et. al.,  Int. J. Biol. Veterin. Agric. Food Engin.,2014, 8, 212-218.