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.2024 Aug 13;10(16):e36185.
doi: 10.1016/j.heliyon.2024.e36185. eCollection 2024 Aug 30.

In vitro evaluation of oral contraceptives on long-tailed macaque (Macaca fascicularis) primary ovarian cells

Affiliations

In vitro evaluation of oral contraceptives on long-tailed macaque (Macaca fascicularis) primary ovarian cells

Mikail Muhammed et al. Heliyon..

Abstract

Hormonal contraception has been advocated as an alternative population control method for the long-tailed macaque population, which has increased exponentially due to anthropogenic changes and incidental food subsidies from human food waste. Risks of increased zoonosis and conflict are imminent if the population growth of long-tailed macaques is unchecked. However, there's a gap in the literature about the effect of hormonal contraceptives on long-tailed macaque reproductive tissues cell line. The present study aims to investigate the effect of oral contraceptives (Nordette, Noriday, and Ella) on long-tailed macaque ovarian cells. We determine the cell viability and cytotoxicity as well as the morphological changes of the drugs on long-tailed macaque ovarian cells using the MTT assay, Acridine orange/propidium iodide double staining method, morphological examination, and the 4, 6-diamidino-2-phenylindole (DAPI) staining method. For the MTT assay, The drugs were dissolved in culture media before use to have a concentration ranging from 0.5 μg/mL, 2.5 μg/mL, 0.125 μg/mL, 0.0625 μg/mL, and 0.0315 μg/mL to have three replicates for each treatment. In contrast, the concentration of 0.0315 μg/mL was used for the morphological and histopathological analysis. The result of the study indicates that human oral contraceptives (Nordette, Noriday, and Ella) inhibit the growth of long-tailed macaque ovarian cells and induce apoptosis in a concentration- and time-dependent manner (at a concentration of 0.0315 μg/mL and an IC50 lower than 10 μg/mL), With a statistically significant value of ****P < 0.001 for each drug compared to the negative control. The result of the present study contributes toward addressing the gap in the literature on the effect of oral contraceptives in long-tailed macaque ovarian cells. Hence, we conclude that human oral contraceptives (Nordette, Noriday, and Ella) are safe and effective in long-tailed macaque ovarian cells as such could be used to develop non-invasive oral contraceptives for controlling the population of long-tailed macaques as an alternative population control method.

Keywords: Apoptosis; Cytotoxicity; Good health and well-being; Long-tailed macaque contraception; Oral contraceptives; Ovarian cells.

© 2024 The Authors. Published by Elsevier Ltd.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Illustration of the procedures for Apoptotic evaluation using Acridine orange/Propidium Iodide (AO/PI), DAPI staining method (Fluorescence microscopy) and morphological evaluation (inverted microscopy). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Fig. 2
Fig. 2
(24h): In vitro cytotoxicity study of long-tailed macaque ovarian cells after 24 h of treatment with Nordette, Noriday, and Ella. Note A value of *P < 0.05; **P < 0.01; ****P < 0.001 is set as statistically significant for each drug compared with the negative control, while ns is not significant when compared within various concentrations of each drug. Fig. 2(48h): In vitro cytotoxicity study of long-tailed macaque ovarian cells after 48 h of treatment with Nordette, Noriday, and Ella. Note A value of *P < 0.05; **P < 0.01; ****P < 0.001 is set as statistically significant for each drug compared with the negative control, while ns is not significant when compared within various concentrations of each drug. Fig. 2(72h): In vitro cytotoxicity study of long-tailed macaque ovarian cells after 72 h of treatment with Nordette, Noriday, and Ella. Note A value of *P < 0.05; **P < 0.01; ****P < 0.001 is set as statistically significant for each drug compared with the negative control, while ns is not significant when compared within various concentrations of each drug.
Fig. 3
Fig. 3
Inverted light microscopy images of long-tailed macaque ovarian cells treated for 48 h with 0.03125 μg/mL (IC50) concentration of human oral contraceptives (A) Control cell(untreated); (B) cells treated with Nordette(Combined oral contraceptives); (C) Cells treated with Noriday (Progestin-only pills); (D) Cells treated with Ella (Progesterone antagonist). Note V.C. arrow denotes viable cells, A.C. arrow denotes apoptotic cells, and N.C. arrow denotes necrotic cells.
Fig. 4
Fig. 4
The fluorescence photomicrographs showing the apoptotic effect of human oral contraceptives on the nuclei morphology of long-tailed macaque ovarian cells, using DAPI Staining. Ovarian cells were treated for 48hrs with 0.03125 μg/mL (IC50) concentration of human oral contraceptives. (A) Control cells (untreated cells); (B) cells treated with Nordette (combined oral contraceptives), (C) cells treated with Noriday (Progestin-only pills), (D) cells treated with Ella (Progesterone antagonist). Note CC arrow denotes chromatin condensation; the N.F. arrow denotes Nuclei fragmentation.
Fig. 5
Fig. 5
The fluorescent photomicrographs showing the human oral contraceptives induced apoptosis on the long-tailed macaque ovarian cells, assessed by Acridine Orange/Propidium iodide (AO/PI) double staining methods. Ovarian cells were treated for 48hrs with 0.03125 μg/mL (IC50) concentration of human oral contraceptives. (A) Control cells (untreated cells); (B) cells treated with Nordette (combined oral contraceptives), (C) cells treated with Noriday (Progestin-only pills), (D) cells treated with Ella (Progesterone antagonist). Note V.C. arrow denotes viable cells, the N.C. arrow denotes Necrotic cells, and the A.C. arrow denotes apoptotic cells. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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References

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