Mechanochemical preparation of photothermal agent based on casein-capped CuS nanoparticles proposed by Taguchi design of experiments Martin Stahorský 1, 2 , Martin Kello 3 , Nina Daneu 4 , Erika Dutková 1 , Zdenka Lukáčová Bujňáková 1 and MatejBaláž 1 1 Department of Mechanochemistry, Institute of Geotechnics, Slovak Academy of Sciences, Slovakia, 2 Faculty of Materials, Metallurgy and Recycling, Technical University of Košice, Slovakia, 3 Department of Pharmacology, Pavol Jozef Šafárik University, Slovakia, 4 Advanced Materials Department, Jožef Stefan Institute, Slovenia In this paper, the biocompatible nanosuspension of CuS nanoparticles capped with casein was prepared using a two-stage mechanochemical approach. CuS nanoparticles were firstly synthesized by a planetary ball milling using elemental precursors [1]. Subsequently, the Design of Experiments (DoE), namely the Taguchi method was applied to determine the optimal conditions for preparation CuS-casein nanosuspension in the process of wet stirred media milling. Optimized factors encompassed the amount of nanoparticles, casein solution concentration and milling time. Each of these factors was changed on three levels. The output data were the average hydrodynamic diameter (D 50 ) of the designed nanosuspensions. For a wet stirred media milling of CuS nanoparticles in a solution of casein, a sample mass of 7 g CuS NPs, casein solution 1% wt.% and milling time of 75 min were found to be optimum. Furthermore, the influence of the centrifugation speed after milling on the average hydrodynamic diameter (D 50 ) and on the copper concentration of the resulting nanosuspension was studied in order to achieve values suitable for biomedical application. The obtained nanosuspension and its stability were characterized by laser diffraction and zeta potential measurements. The nanocrystalline character of CuS and the presence of casein were confirmed by UV–Vis, PL and FT-IR spectroscopy, transmission electron microscopy and X-ray diffraction. To test the suitability of the prepared CuS-casein nanosuspension for photothermal ablation application, cancer cell lines (MCF-7, A549, HeLa, HCT116 and BLM) and non-cancer cell line (COS-7) containing our CuS-casein nanocrystals were irradiated with near-infrared radiation for 5 and 10 min. Photothermal destruction was induced upon an irradiation in a dose- dependent, as well as nanoparticles concentration-dependent manner. The synthesized CuS-casein nanocrystals prepared by us are among suitable candidates for photothermal therapy of cancer. The contribution was supported by the Slovak Research and Development Agency (contract No. APVV-18-0357), the Slovak Grant Agency of the Ministry of Education, Science, Research and Sports of the Slovak Republic (VEGA No. 2/0112/22), and the Slovak Academy of Sciences from the Program of Grants for Doctoral Students (DoktoGrant APP0330). References 1. Baláž M., Zorkovská A., Blazquez J. S., Daneu N., Baláž P. Mechanochemistry of copper sulphides: phase interchanges during milling // J. Mater. Sci.-2017.- 52 , N 20.-P. 11947-11961.
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