Nickel oxide nanoparticles exposure as a risk factor for male infertility: “In vitro” effects on porcine pre-pubertal Sertoli cells

Abstract

Lately, nickel oxide nanoparticles (NiO NPs) have been employed in different industrial and biomedical fields. Several studies have reported that NiO NPs may affect the development of reproductive organs inducing oxidative stress and, resulting in male infertility. We investigated the in vitro effects of NiO NPs on porcine pre-pubertal Sertoli cells (SCs) which undergone acute (24 h) and chronic (from 1 up to 3 weeks) exposure at two subtoxic doses of NiO NPs of 1 μg/ml and 5 μg/ml. After NiO NPs exposure we performed the following analysis: (a) SCs morphological analysis (Light Microscopy); (b) ROS production and oxidative DNA damage, gene expression of antioxidant enzymes (c) SCs functionality (AMH, inhibin B Real-time PCR analysis and ELISA test); (d) apoptosis (WB analysis); (e) pro-inflammatory cytokines (Real-time PCR analysis), and (f) MAPK kinase signaling pathway (WB analysis). We found that the SCs exposed to both subtoxic doses of NiO NPs didn’t sustain substantial morphological changes. NiO NPs exposure, at each concentration, reported a marked increase of intracellular ROS at the third week of treatment and DNA damage at all exposure times. We demonstrated, un up-regulation of SOD and HO-1 gene expression, at both concentrations tested. The both subtoxic doses of NiO NPs detected a down-regulation of AMH and inhibin B gene expression and secreted proteins. Only the 5 μg/ml dose induced the activation of caspase-3 at the third week. At the two subtoxic doses of NiO NPs a clear pro-inflammatory response was resulted in an up-regulation of TNF-α and IL-6 in terms of mRNA. Finally, an increased phosphorylation ratio of p-ERK1/2, p-38 and p-AKT was observed up to the third week, at both concentrations. Our results show the negative impact of subtoxic doses NiO NPs chronic exposure on porcine SCs functionality and viability.

Oxidative Stress, Cytotoxic and Inflammatory Effects of Azoles Combinatorial Mixtures in Sertoli TM4 Cells

“…As pivotal elements for an adequate development of testis and the spermatogenesis process, SCs are a commonly used cellular model for testing cytotoxicity and its relationship with male reproductive system dysfunctions, since they have been ascertained to be the cellular target of many environmental and chemical toxicants [ 54 ]. As a consequence, many conditions of male subfertility or infertility might be due to SCs impairment [ 55 , 56 , 57 , 58 , 59 , 60 , 61 ], potentially subsisting due to exposure to a plethora of xenobiotics and chemical mixtures in the daily living environment [ 62 , 63 ]. Nowadays, emerging toxicant-mediated mechanisms contributing to male reproductive dysfunctions often reveal OS conditions and ROS over-production [ 64 , 65 ].…”

“…Additionally, folpet-induced OS mediates antiproliferative effects and apoptotic cell death. Additionally, porcine pre-pubertal SCs exposed to subtoxic doses of nickel oxide nanoparticles (NiO NPs) showed a marked increase in intracellular ROS, DNA damage, activation of caspase-3, and a clear pro-inflammatory stress that resulted in an up-regulation of TNF-α and IL-6 [ 56 ]. Endocrine disruptors such as bisphenol A (BPA) induce OS, which affects sperm functions [ 65 ].…”