“…Therefore, in order to collect information on primary granulosa cells we isolated and cells from the pig, which represents a reliable animal model due to its well-known value in translational medicine [ 28 ]. We studied the effects produced by Triclosan on the main functional activity of granulosa cells, i.e., growth, steroidogenesis and redox status markers [ 27 ], after selecting the concentrations on the bases of previous reports [ 14 , 24 , 35 ]. The Triclosan tested concentrations were chosen on the basis of the assays in human matrices.…”
Section: Discussionmentioning
confidence: 99%
“…Cell number/well was estimated from the resulting linear regression equation and was used to correct experimental data. The assay detection limit was 10 3 cell/well and the variation coefficient was less than 5% [ 27 ].…”
Section: Methodsmentioning
confidence: 99%
“…A total of 2 × 10 5 cells/100 µL CM were placed in 96-well plates and treated with Triclosan as detailed above. Luminescence was measured using a Victor Reader after kit reagent addition [ 27 ].…”
Section: Methodsmentioning
confidence: 99%
“…In particular, the present research was performed to deepen the study of the potential effects on ovarian function. Thus, granulosa cells were isolated from swine ovarian follicles, accordingly to a validated method [ 25 , 26 , 27 ]. It is well known that the pig represents an excellent model for translational medicine due to physiological similarities with the human [ 28 ].…”
Triclosan is a chlorinated biphenolic with a broad spectrum of antiseptic activities used in cosmetics and hygiene products. Continuous exposure can lead to the absorption and bioaccumulation of this substance with harmful health effects. In fact, previous studies have shown that Triclosan acts as an endocrine-disrupting chemical on reproductive organs, with consequent negative effects on reproductive physiology. Therefore, to assess potential adverse impacts on fertility, we tested Triclosan on swine granulosa cells, a model of endocrine reproductive cells. We examined its effects on the main features of granulosa cell functions such as cell growth (BrdU incorporation and ATP production) and steroidogenesis (17-β estradiol and progesterone secretion). Moreover, since oxidant–antioxidant balance plays a pivotal role in follicular function, redox status markers (superoxide, hydrogen peroxide and nitric oxide production, enzymatic and non-enzymatic scavenging activity) were studied. Our results show that Triclosan significantly inhibits cell growth (p < 0.001), steroidogenesis (p < 0.001), superoxide and nitric oxide production (p < 0.001), while it increases (p < 0.05) enzymatic defense systems. Collectively, these data suggest a disruption of the main granulosa cell functions, i.e., proliferation and hormone production, as well as and an imbalance in redox status. On these bases, we can speculate that Triclosan would impair granulosa cell functions, thus exerting negative effects on reproductive function. Further studies are needed to explore lower Triclosan concentrations and to unravel its mechanisms of action at gene level.
“…Therefore, in order to collect information on primary granulosa cells we isolated and cells from the pig, which represents a reliable animal model due to its well-known value in translational medicine [ 28 ]. We studied the effects produced by Triclosan on the main functional activity of granulosa cells, i.e., growth, steroidogenesis and redox status markers [ 27 ], after selecting the concentrations on the bases of previous reports [ 14 , 24 , 35 ]. The Triclosan tested concentrations were chosen on the basis of the assays in human matrices.…”
Section: Discussionmentioning
confidence: 99%
“…Cell number/well was estimated from the resulting linear regression equation and was used to correct experimental data. The assay detection limit was 10 3 cell/well and the variation coefficient was less than 5% [ 27 ].…”
Section: Methodsmentioning
confidence: 99%
“…A total of 2 × 10 5 cells/100 µL CM were placed in 96-well plates and treated with Triclosan as detailed above. Luminescence was measured using a Victor Reader after kit reagent addition [ 27 ].…”
Section: Methodsmentioning
confidence: 99%
“…In particular, the present research was performed to deepen the study of the potential effects on ovarian function. Thus, granulosa cells were isolated from swine ovarian follicles, accordingly to a validated method [ 25 , 26 , 27 ]. It is well known that the pig represents an excellent model for translational medicine due to physiological similarities with the human [ 28 ].…”
Triclosan is a chlorinated biphenolic with a broad spectrum of antiseptic activities used in cosmetics and hygiene products. Continuous exposure can lead to the absorption and bioaccumulation of this substance with harmful health effects. In fact, previous studies have shown that Triclosan acts as an endocrine-disrupting chemical on reproductive organs, with consequent negative effects on reproductive physiology. Therefore, to assess potential adverse impacts on fertility, we tested Triclosan on swine granulosa cells, a model of endocrine reproductive cells. We examined its effects on the main features of granulosa cell functions such as cell growth (BrdU incorporation and ATP production) and steroidogenesis (17-β estradiol and progesterone secretion). Moreover, since oxidant–antioxidant balance plays a pivotal role in follicular function, redox status markers (superoxide, hydrogen peroxide and nitric oxide production, enzymatic and non-enzymatic scavenging activity) were studied. Our results show that Triclosan significantly inhibits cell growth (p < 0.001), steroidogenesis (p < 0.001), superoxide and nitric oxide production (p < 0.001), while it increases (p < 0.05) enzymatic defense systems. Collectively, these data suggest a disruption of the main granulosa cell functions, i.e., proliferation and hormone production, as well as and an imbalance in redox status. On these bases, we can speculate that Triclosan would impair granulosa cell functions, thus exerting negative effects on reproductive function. Further studies are needed to explore lower Triclosan concentrations and to unravel its mechanisms of action at gene level.
“…An amount of 4 × 10 4 cells/100 µL of CM was seeded in 96-well plates and treated with Irisin as detailed above. The assay was performed according to the instructions, and luminescence was finally recorded by the Victor Nivo luminometer [ 14 ].…”
Irisin is a hormone able to reproduce some of the positive effects of physical activity and diet. Recently, we demonstrated the presence of Irisin at the ovarian level as a potential physiological regulator of follicular function. Adipose tissue is crucial for reproductive function through its metabolic activity and the production of adipokines. At present, the exact nature of adipocyte precursors is still under debate, but an important role has been assigned to the population of adipose tissue mesenchymal stromal cells (ASCs) of perivascular origin. It should be noted that, when appropriately stimulated, ASCs can differentiate into preadipocytes and, subsequently, adipocytes. Therefore, this present study was undertaken to explore the potential effect of Irisin on ASCs, known for their high differentiative potential. Since Irisin expression in ASCs was confirmed by PCR, we tested its potential effects on the main functional activities of these cells, including proliferation (BrdU uptake); metabolic activity (ATP production); redox status, evaluated as the generation of free molecules such as superoxide anion and nitric oxide; and scavenger activities, assessed as both enzymatic (superoxide dismutase) and non-enzymatic antioxidant power. Moreover, we tested the effect of Irisin on ASCs adipogenic differentiation. BrdU uptake was significantly (p < 0.001) inhibited by Irisin, while ATP production was significantly (p < 0.05) increased. Both superoxide anion and nitric oxide generation were significantly increased (p < 0.001) by Irisin, while scavenger activity was significantly reduced (p < 0.05). Irisin was found to significantly (p < 0.05) inhibit ASCs adipogenic differentiation. Taken together, the present results suggest a potential local role of Irisin in the regulation of adipose tissue function.
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