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Biological Application of a Fluorescent Zinc Sensing Probe for the Analysis of Zinc Bioavailability Using Caco-2 Cells as an In-Vitro Cellular Model

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Abstract

Zinc is essential for growth and development of all living organisms, especially human being. Deficiency of micronutrients like zinc and iron has been linked to the manifestation of hidden hunger. Therefore, it is imperative that development of some rapid screening method for bioavailable zinc in various crops and food commodities would be an essential addition in battle against zinc deficiency related hidden hunger. One such method could be the usage of fluorescence based zinc ion sensing probe which would be robust and convenient for estimating bioavailable zinc. To address this issue, NBD-TPEA, a highly sensitive zinc ion sensing probe, have been used in this study towards the development of a novel fluorescence based approach for the analysis of zinc bioavailability in Caco-2 cells as an in-vitro cellular model. The use of this probe showed dose dependent sensitivity towards increasing concentrations of zinc ion uptake by Caco-2 cells. It also showed specificity for zinc ion uptake as compared to other metal ions in-vitro. These observations correlated extremely well with zinc uptake analysis by cell imaging and conventional analytical technique like, ICP-MS. The developed assay was then tested in mushroom and some selected biofortified derivatives of wheat for determining the levels of their bioavailable zinc using Caco-2 cells. The data as obtained with these food samples in our developed bioassay correlated well with the other sophisticated analytical techniques thus validating our cell based assay. Hence, the developed assay could serve as a simple but sensitive tool for determining bioavailable zinc in various food samples.

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Acknowledgements

The authors would also like to thank Department of Biotechnology, Ministry of Science and Technology, Government of India for providing financial support to PR and fellowship to SN (File No. BT/AGR/biofort/PHI/NIN/2011). The authors would like to thank the technical assistance of Institute Instrumentation Centre at Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India for various experiments.

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SN and PR designed the study. SN completed the experimental process, literature search and generation of figures. VK and HSD provided the wheat samples. SN and PR wrote the manuscript while VK, HSD and DS reviewed and edited the manuscript. All authors read and approved the final manuscript.

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Correspondence to Partha Roy.

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Nathani, S., Kumar, V., Dhaliwal, H.S. et al. Biological Application of a Fluorescent Zinc Sensing Probe for the Analysis of Zinc Bioavailability Using Caco-2 Cells as an In-Vitro Cellular Model. J Fluoresc 30, 1553–1565 (2020). https://doi.org/10.1007/s10895-020-02608-w

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