Perfluorooctanoic Acid (PFOA) Induces Redox Status Disruption in Swine Granulosa Cells

Abstract

Perfluorooctanoic acid (PFOA) is employed in the production and processing of several plastic materials, mainly during the production of waterproof fabrics or nonstick cookware. PFOA is identified as a substance of very high concern, as it is classified as a persistent, bioaccumulative, and toxic (PBT) substance because of its persistence in the environment and its potential accumulation in organisms. Thus, safe levels of exposure cannot be established, and PFOA emissions should be minimized. PFOA has recently been linked to several health concerns in humans. In particular, a disruptive effect on redox status homeostasis has been documented, with a potential impairment of normal reproductive function that requires adequate oxidative balance. Therefore, the aim of the present study was to evaluate the effects of PFOA (2, 20, and 200 ng/mL) on ovarian granulosa cells, a model of reproductive cells. The obtained results reveal that PFOA stimulated cell viability (p < 0.05). Regarding the effects on free radical production, O2−, NO, and H2O2 were significantly inhibited (p < 0.05), while the nonenzymatic antioxidant power was not significantly modified. Collectively, the present results deserve attention since free radical molecules play a crucial role in ovarian follicle development leading to a successful ovulation.

Keywords: PFAS; cell viability; free radicals; ovary; oxidative stress.

Figure 1

Effect of 48 h treatment with or without (C) perfluorooctanoic acid (PFOA) (2, 20 and 200 ng/mL) on cell metabolic activity quantified using ATP production in swine granulosa cell culture media. Data expressed as counts per second (CPS) represent the mean ± SEM of six replicates/treatment repeated in five different experiments. Different letters indicate a significant difference (p < 0.05).

Figure 2

Effect of 48 h treatment with or without (C) perfluorooctanoic acid (PFOA) (2, 20 and 200 ng/mL) on hydrogen peroxide (H₂O₂) production in swine granulosa cell lysates. Data expressed as µM represent the mean ± SEM of six replicates/treatment repeated in five different experiments. Different letters indicate a significant difference (p < 0.05).

Figure 3

Effect of 48 h treatment with or without (C) perfluorooctanoic acid (PFOA) (2, 20, and 200 ng/mL) on superoxide anion (O₂⁻) generation in swine granulosa cell culture media. Data expressed as milliabsorbance units (milliAbs), represent the mean ± SEM of six replicates/treatment repeated in five different experiments. Different letters indicate a significant difference (p < 0.05).

Figure 4

Effect of 48 h treatment with or without (C) perfluorooctanoic acid (PFOA) (2, 20 and 200 ng/mL) on NO production quantified using Griess reagent in swine granulosa cell culture media. Data expressed as µM represent the mean ± SEM of six replicates/treatment repeated in five different experiments. Different letters indicate a significant difference (p < 0.05).

Figure 5

Effect of 48 h treatment with or without (C) perfluorooctanoic acid (PFOA) (2, 20 and 200 ng/mL) on nonenzymatic scavenger activity quantified using FRAP method in swine granulosa cell culture media. Data, expressed as µM, represent the mean ± SEM of six replicates/treatment repeated in five different experiments.