Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells

Copper nanoparticles (CuNPs) are known to affect many ovarian cell functions. CuNPs, prepared using a chemical reduction method, were fully characterized by different means (TEM, DLS, XRD, Z potential, XPS, and AES). The resulting colloidal suspension contained needle-like CuNPs aggregates made of a...

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Main Authors: Alexander V. Sirotkin, Paula Romero-Navarro, Barbora Loncová, Zuzana Fabová, Michaela Bartušová, Abdel Halim Harrath, Francisco Alonso
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Nanomaterials
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Online Access:https://www.mdpi.com/2079-4991/14/23/1965
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author Alexander V. Sirotkin
Paula Romero-Navarro
Barbora Loncová
Zuzana Fabová
Michaela Bartušová
Abdel Halim Harrath
Francisco Alonso
author_facet Alexander V. Sirotkin
Paula Romero-Navarro
Barbora Loncová
Zuzana Fabová
Michaela Bartušová
Abdel Halim Harrath
Francisco Alonso
author_sort Alexander V. Sirotkin
collection DOAJ
description Copper nanoparticles (CuNPs) are known to affect many ovarian cell functions. CuNPs, prepared using a chemical reduction method, were fully characterized by different means (TEM, DLS, XRD, Z potential, XPS, and AES). The resulting colloidal suspension contained needle-like CuNPs aggregates made of a core of metallic copper and an oxidized surface of Cu<sub>2</sub>O and CuO. The separate and coupled effects of CuNPs and the growth factor betacellulin (BTC) were analyzed on the control of some basic functions of ovarian cells. With this purpose, porcine ovarian granulosa cells, together with CuNPs, BTC, and both (CuNPs + BTC), were cultured. Viability and BrDU tests, quantitative immunocytochemistry, TUNEL, and ELISA were used to evaluate markers of the S-phase (PCNA) and G-phase (cyclin B1) of the cell cycle, cell proliferation (BrDU incorporation), cytoplasmic/mitochondrial apoptosis (bax) and extrinsic (nuclear DNA fragmentation) markers, and the release of estradiol and progesterone. CuNPs were accumulated within the cells and were found to reduce all the markers of proliferation, but promoted all the markers of apoptosis and the release of steroid hormones. When added alone, BTC raised the expression of all cell viability and proliferation markers, depleted the expression of all apoptosis markers, and stimulated the release of both estradiol and progesterone. Furthermore, BTC prevented and even reversed the effect of CuNPs on all the measured parameters, whereas CuNPs mitigated BTC’s effect on all the analyzed cell functions. These results support a direct toxic effect of CuNPs and a stimulatory effect of BTC on ovarian cell functions, as well as the capability of BTC to protect against the adverse effects of CuNPs.
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spelling doaj-art-c577853794284f76a6cd6f88e2e2b00b2025-08-20T02:50:40ZengMDPI AGNanomaterials2079-49912024-12-011423196510.3390/nano14231965Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian CellsAlexander V. Sirotkin0Paula Romero-Navarro1Barbora Loncová2Zuzana Fabová3Michaela Bartušová4Abdel Halim Harrath5Francisco Alonso6Department of Zoology and Anthropology, Constantine the Philosopher University, Tr. A Hlinku 1, 949 74 Nitra, SlovakiaInstituto de Síntesis Orgánica (ISO) and Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Alicante, Apdo. 99, 03080 Alicante, SpainDepartment of Zoology and Anthropology, Constantine the Philosopher University, Tr. A Hlinku 1, 949 74 Nitra, SlovakiaDepartment of Zoology and Anthropology, Constantine the Philosopher University, Tr. A Hlinku 1, 949 74 Nitra, SlovakiaDepartment of Zoology and Anthropology, Constantine the Philosopher University, Tr. A Hlinku 1, 949 74 Nitra, SlovakiaDepartment of Zoology, College of Science, King Saud University, Riyadh 11451, Saudi ArabiaInstituto de Síntesis Orgánica (ISO) and Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Alicante, Apdo. 99, 03080 Alicante, SpainCopper nanoparticles (CuNPs) are known to affect many ovarian cell functions. CuNPs, prepared using a chemical reduction method, were fully characterized by different means (TEM, DLS, XRD, Z potential, XPS, and AES). The resulting colloidal suspension contained needle-like CuNPs aggregates made of a core of metallic copper and an oxidized surface of Cu<sub>2</sub>O and CuO. The separate and coupled effects of CuNPs and the growth factor betacellulin (BTC) were analyzed on the control of some basic functions of ovarian cells. With this purpose, porcine ovarian granulosa cells, together with CuNPs, BTC, and both (CuNPs + BTC), were cultured. Viability and BrDU tests, quantitative immunocytochemistry, TUNEL, and ELISA were used to evaluate markers of the S-phase (PCNA) and G-phase (cyclin B1) of the cell cycle, cell proliferation (BrDU incorporation), cytoplasmic/mitochondrial apoptosis (bax) and extrinsic (nuclear DNA fragmentation) markers, and the release of estradiol and progesterone. CuNPs were accumulated within the cells and were found to reduce all the markers of proliferation, but promoted all the markers of apoptosis and the release of steroid hormones. When added alone, BTC raised the expression of all cell viability and proliferation markers, depleted the expression of all apoptosis markers, and stimulated the release of both estradiol and progesterone. Furthermore, BTC prevented and even reversed the effect of CuNPs on all the measured parameters, whereas CuNPs mitigated BTC’s effect on all the analyzed cell functions. These results support a direct toxic effect of CuNPs and a stimulatory effect of BTC on ovarian cell functions, as well as the capability of BTC to protect against the adverse effects of CuNPs.https://www.mdpi.com/2079-4991/14/23/1965apoptosisbetacellulincoppernanoparticlesovaryproliferation
spellingShingle Alexander V. Sirotkin
Paula Romero-Navarro
Barbora Loncová
Zuzana Fabová
Michaela Bartušová
Abdel Halim Harrath
Francisco Alonso
Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells
Nanomaterials
apoptosis
betacellulin
copper
nanoparticles
ovary
proliferation
title Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells
title_full Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells
title_fullStr Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells
title_full_unstemmed Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells
title_short Counteractive Effects of Copper Nanoparticles and Betacellulin on Ovarian Cells
title_sort counteractive effects of copper nanoparticles and betacellulin on ovarian cells
topic apoptosis
betacellulin
copper
nanoparticles
ovary
proliferation
url https://www.mdpi.com/2079-4991/14/23/1965
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