Effect of morphology and support of copper nanoparticles on basic ovarian granulosa cell functions

The aim of this survey is to explore the possible effects of unsupported and supported copper nanoparticles (CuNPs) of different morphologies on basic ovarian cell functions. For this purpose, we have compared the activity of unsupported spherical, triangular, and hexagonal CuNPs, as well as of spherical CuNPs supported on titania, zeolite Y and activated charcoal (0, 1, 10, or 100 ng/mL) on cultured porcine ovarian granulosa cells. Cell viability, proliferation (accumulation of proliferating cell nuclear antigen, PCNA), apoptosis (accumulation of Bcl-2-associated X protein, bax) and release of steroid hormones progesterone, testosterone, and 17β-estradiol have been analyzed by the Trypan blue test, quantitative immunocytochemistry, and ELISA, respectively. Cell viability decreased after treatment with hexagonal CuNPs, whilst all the other CuNPs increased it. Unsupported spherical and hexagonal CuNPs, and spherical CuNPs/titania reduced PCNA accumulation; in contrast, an increase was noted for unsupported triangular CuNPs and CuNPs/zeolite Y. Bax accumulation was not affected by hexagonal CuNPs, whereas CuNPs/zeolite Y promoted it and all the other CuNPs depleted it. The release of all steroid hormones was inhibited by CuNPs/titanium dioxide and stimulated by CuNPs/charcoal, whilst CuNPs/zeolite Y promoted the testosterone and 17β-estradiol output, but not that of progesterone. These results demonstrate the direct, mainly stimulatory, impact of CuNPs on basic ovarian cell functions. The character of the CuNPs' action depends on their shape and support. Therefore, CuNPs with appropriate chemical modification could be potentially useful for the control of reproductive processes and treatment of reproductive disorders.This study was supported by the Slovak Research and Development Agency [project no. APVV-15-0296], the Slovak Grant Agency of the Ministry of Education, Science and Sport, the Slovak Academy of Science [project no. VEGA 1/0392/17], Spanish Ministerio de Ciencia, Innovación y Universidades [project no. CTQ2017-88171-P], the Generalitat Valenciana [project no. AICO/2017/007], and the Instituto de Síntesis Orgánica. I.M.-G. thanks the Vicerrectorado de Investigación y Transferencia del Conocimiento of the Universidad de Alicante for a pre-doctoral grant [no. UAFPU2016-034]