{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T19:06:06Z","timestamp":1792263966089,"version":"4.4.0"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,2,23]],"date-time":"2024-02-23T00:00:00Z","timestamp":1708646400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Bioeng. Biotechnol."],"abstract":"<jats:p><jats:bold>Introduction:<\/jats:bold> The development of an effective extender is important for semen preservation and the artificial insemination (AI) industry. This study demonstrates the beneficial effect of zinc oxide nanoparticles (ZnO-NPs) as an additive to semen extenders to improve semen quality, fertility, and antibacterial activity during liquid preservation in a boar model.<\/jats:p><jats:p><jats:bold>Methods:<\/jats:bold> Initially, to find out the safe concentration of ZnO-NPs in sperm cells, a wide range of ZnO-NP concentrations (0, 5, 10, 50, 100, 500, and 1,000\u00a0\u03bcM) were co-incubated with sperm at 37\u00b0C for a cytotoxic study. These NP concentrations were compared to their salt control zinc acetate (ZA) at the same concentrations and to a control group. The effect of the different concentrations of ZnO-NPs on sperm motility, membrane integrity, mitochondrial membrane potential (MMP), and apoptosis was assessed. Accordingly, the non-toxic dose was selected and supplemented in MODENA extender to determine its beneficial effect on the boar semen parameters mentioned and the lipid peroxidation (LPO) levels during liquid preservation at 16\u00b0C for 6\u00a0days. The non-cytotoxic dosage was subsequently chosen for AI, fertility investigations, and the evaluation of the antibacterial efficacy of ZnO-NPs during preservation hours. An antibacterial study of ZnO-NPs and its salt control at doses of 10\u00a0\u03bcM and 50\u00a0\u03bcM was carried out by the colony forming unit (CFU) method.<\/jats:p><jats:p><jats:bold>Results and discussion:<\/jats:bold> The cytotoxic study revealed that 5, 10, and 50\u00a0\u03bcM of ZnO-NPs are safe. Consequently, semen preserved in the MODENA extender, incorporating the non-toxic dose, exhibited 10 and 50\u00a0\u03bcM ZnO-NPs as the optimal concentrations for beneficial outcomes during liquid preservation at 16\u00b0C. ZnO-NPs of 10\u00a0\u03bcM concentration resulted in a significantly (<jats:italic>p<\/jats:italic> &amp;lt; 0.05) improved conception rate of 86.95% compared to the control of 73.13%. ZnO-NPs of 10 and 50\u00a0\u03bcM concentrations exhibit potent antimicrobial action by reducing the number of colonies formed with days of preservation in comparison to the negative control. The investigation concluded that the incorporation of 10\u00a0\u03bcM ZnO-NPs led to enhancements in sperm motility, membrane integrity, and MMP, attributed to a reduction in the malondialdehyde (MDA) levels. This improvement was accompanied by a concurrent increase in fertility rates, including farrowing rate and litter size, during the liquid preservation process. Furthermore, ZnO-NPs exhibited an antimicrobial effect, resulting in decreased bacterial growth while preserving boar semen at 16\u00b0C for 6\u00a0days. These findings suggest that ZnO-NPs could serve as a viable alternative to antibiotics, potentially mitigating antibiotic resistance concerns within the food chain.<\/jats:p>","DOI":"10.3389\/fbioe.2024.1326143","type":"journal-article","created":{"date-parts":[[2024,2,23]],"date-time":"2024-02-23T05:01:57Z","timestamp":1708664517000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":13,"title":["Synthesis of green zinc\u2010oxide nanoparticles and its dose\u2010dependent beneficial effect on spermatozoa during preservation: sperm functional integrity, fertility and antimicrobial activity"],"prefix":"10.3389","volume":"12","author":[{"given":"Meiaishan Eliezer","family":"Lyngdoh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jyoti","family":"Chettri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivian F.","family":"Kharchandy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rishav","family":"Sheel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arnab Roy","family":"Choudhury","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biplab","family":"Sarkar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arunava","family":"Pattanayak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sourabh","family":"Deori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sayed Nabil","family":"Abedin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Kadirvel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,2,23]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1508","DOI":"10.3390\/ani10091508","article-title":"The effect of adding different levels of curcumin and its nanoparticles to extender on post-thaw quality of cryopreserved rabbit sperm","volume":"10","author":"Abdelnour","year":"2020","journal-title":"Anim. (Basel)"},{"key":"B2","doi-asserted-by":"publisher","first-page":"127296","DOI":"10.1016\/j.jtemb.2023.127296","article-title":"Zinc oxide and selenium nanoparticles can improve semen quality and heat shock protein expression in cryopreserved goat (Capra hircus) spermatozoa","volume":"80","author":"Abedin","year":"","journal-title":"J. Trace Elem. Med. Biol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"4726","DOI":"10.1007\/s12011-022-03551-6","article-title":"In vitro and in vivo studies on the efficacy of zinc-oxide and selenium nanoparticle in cryopreserved goat (Capra hircus) spermatozoa","volume":"201","author":"Abedin","year":"","journal-title":"Biol. Trace Elem. Res."},{"key":"B4","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2015\/153573","article-title":"Ameliorative effect of zinc oxide nanoparticles on antioxidants and sperm characteristics in streptozotocin-induced diabetic rat testes","volume":"2015","author":"Afifi","year":"2015","journal-title":"Biomed. Res. Int."},{"key":"B5","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1095\/biolreprod41.1.183","article-title":"Generation of reactive oxygen species, lipid peroxidation, and human sperm function","volume":"41","author":"Aitken","year":"1989","journal-title":"Biol. Reprod."},{"key":"B6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.4061\/2011\/686137","article-title":"Impacts of oxidative stress and antioxidants on semen functions","volume":"2011","author":"Bansal","year":"2010","journal-title":"Vet. Med. Int."},{"key":"B7","first-page":"767","article-title":"Effect of zinc oxide nanoparticles on viability of human spermatozoa","volume":"11","author":"Barkhordari","year":"2013","journal-title":"Iran. J. Reprod. Med."},{"key":"B8","doi-asserted-by":"publisher","first-page":"e13709","DOI":"10.1111\/and.13709","article-title":"Nano\u2010purification of raw semen minimises oxidative stress with improvement in post\u2010thaw quality of buffalo spermatozoa","volume":"52","author":"Bisla","year":"2020","journal-title":"Andrologia"},{"key":"B9","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1016\/j.smallrumres.2009.11.015","article-title":"The effect of antioxidants on post-thawed Angora goat (Capra hircusancryrensis) sperm parameters, lipid peroxidation and antioxidant activities","volume":"89","author":"Bucak","year":"2010","journal-title":"Small Rumin. Res."},{"key":"B10","doi-asserted-by":"publisher","first-page":"302","DOI":"10.1016\/S0076-6879(78)52032-6","article-title":"Microsomal lipid peroxidation","volume":"52","author":"Buege","year":"1978","journal-title":"Meth. Enzymol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1315","DOI":"10.1080\/21691401.2020.1843470","article-title":"Antimicrobial, antioxidant and anticancer activities of gold nanoparticles green synthesized using Mangifera indica seed aqueous extract","volume":"48","author":"Donga","year":"2020","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"B12","first-page":"221","article-title":"Characteristics of sperm motility in boar semen diluted in different extenders and stored for seven days at 18 degrees C","volume":"7","author":"Estienne","year":"2007","journal-title":"Reprod. Biol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"832","DOI":"10.1002\/mrd.22831","article-title":"Novel agents for sperm purification, sorting, and imaging","volume":"84","author":"Feugang","year":"2017","journal-title":"Mol. Reprod. Dev."},{"key":"B14","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1016\/j.theriogenology.2004.09.023","article-title":"Sperm factors related to in vitro and in vivo porcine fertility","volume":"63","author":"Gadea","year":"2005","journal-title":"Theriogenology"},{"key":"B15","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1095\/biolreprod56.3.707","article-title":"Regulation of protein tyrosine phosphorylation during bovine sperm capacitation by a cyclic adenosine 3\u20195\u2019-monophosphate-dependent pathway","volume":"56","author":"Galantino-Homer","year":"1997","journal-title":"Biol. Reprod."},{"key":"B16","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1095\/biolreprod34.1.127","article-title":"Assessment of spermatozoal function using dual fluorescent staining and flow cytometric analyses","volume":"34","author":"Garner","year":"1986","journal-title":"Biol. Reprod."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1401","DOI":"10.1095\/biolreprod57.6.1401","article-title":"Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa","volume":"57","author":"Garner","year":"1997","journal-title":"Biol. Reprod."},{"key":"B18","first-page":"515","article-title":"Efficacy of different extenders on sperm characteristics and fertility in crossbred pigs of north-eastern India","volume":"86","author":"Govindasamy","year":"2016","journal-title":"Vet. Arh."},{"key":"B19","doi-asserted-by":"publisher","first-page":"592","DOI":"10.1016\/j.theriogenology.2014.05.028","article-title":"Treatment with zinc, d-aspartate, and coenzyme Q10 protects bull sperm against damage and improves their ability to support embryo development","volume":"82","author":"Gualtieri","year":"2014","journal-title":"Theriogenology"},{"key":"B20","doi-asserted-by":"publisher","first-page":"71","DOI":"10.9775\/kvfd.2018.20349","article-title":"Nano \u00e7inko oksitin moghani ko\u00e7 semeninin \u00e7\u00f6zd\u00fcrme sonras\u0131 de\u011fi\u015fkenler ve oksidatif durumuna etkisi","volume":"25","author":"Heidari","year":"2018","journal-title":"Kafkas Univ. Vet. Fak. Derg."},{"key":"B21","doi-asserted-by":"publisher","first-page":"656","DOI":"10.1016\/j.bbrc.2017.10.112","article-title":"Supplementing zinc oxide nanoparticles to cryopreservation medium minimizes the freeze-thaw-induced damage to spermatozoa","volume":"494","author":"Isaac","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B22","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1007\/s12011-020-02153-4","article-title":"In vivo and in vitro evaluation of bull semen processed with zinc (Zn) nanoparticles","volume":"199","author":"Jahanbin","year":"2021","journal-title":"Biol. Trace Elem. Res."},{"key":"B23","doi-asserted-by":"publisher","first-page":"249","DOI":"10.3389\/fmats.2020.00249","article-title":"Biogenic synthesis and characterization of antimicrobial and antiparasitic zinc oxide (ZnO) nanoparticles using aqueous extracts of the himalayan columbine (aquilegia pubiflora)","volume":"7","author":"Jan","year":"2020","journal-title":"Front. Mater."},{"key":"B24","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2018\/1062562","article-title":"The advancing of zinc oxide nanoparticles for biomedical applications","volume":"2018","author":"Jiang","year":"2018","journal-title":"Bioinorg. Chem. Appl."},{"key":"B25","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1093\/ps\/82.4.640","article-title":"Alternatives to antibiotics: bacteriocins, antimicrobial peptides and bacteriophages","volume":"82","author":"Joerger","year":"2003","journal-title":"Poult. Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1016\/s0378-4320(00)00157-3","article-title":"Storage of boar semen","volume":"62","author":"Johnson","year":"2000","journal-title":"Anim. Reprod. Sci."},{"key":"B27","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1016\/j.theriogenology.2018.12.017","article-title":"Impact of selenium nano-particles in semen extender on bull sperm quality after cryopreservation","volume":"126","author":"Khalil","year":"2019","journal-title":"Theriogenology"},{"key":"B28","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1016\/j.theriogenology.2015.07.009","article-title":"Artificial insemination in pigs today","volume":"85","author":"Knox","year":"2016","journal-title":"Theriogenology"},{"key":"B29","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1038\/nm1467","article-title":"Artificially engineered magnetic nanoparticles for ultra-sensitive molecular imaging","volume":"13","author":"Lee","year":"2007","journal-title":"Nat. Med."},{"key":"B30","doi-asserted-by":"publisher","first-page":"1319","DOI":"10.3390\/antibiotics10111319","article-title":"Antimicrobially active semen extenders allow the reduction of antibiotic use in pig insemination","volume":"10","author":"Luther","year":"2021","journal-title":"Antibiot. (Basel)"},{"key":"B31","doi-asserted-by":"publisher","first-page":"363","DOI":"10.1016\/bs.coac.2020.12.011","article-title":"Biosynthesized\/green-synthesized nanomaterials as potential vehicles for delivery of antibiotics\/drugs","volume":"94","author":"Mehnath","year":"2021","journal-title":"Compr. Anal. Chem."},{"key":"B32","doi-asserted-by":"publisher","first-page":"1825","DOI":"10.1016\/j.drudis.2017.08.006","article-title":"Zinc oxide nanoparticles: a promising nanomaterial for biomedical applications","volume":"22","author":"Mishra","year":"2017","journal-title":"Drug Discov. Today."},{"key":"B33","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1016\/j.anireprosci.2010.11.005","article-title":"Removal of bacteria from boar ejaculates by Single Layer Centrifugation can reduce the use of antibiotics in semen extenders","volume":"123","author":"Morrell","year":"2011","journal-title":"Anim. Reprod. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"112","DOI":"10.22271\/chemi.2020.v8.i2c.9666","article-title":"Recent advances in preservation of boar semen in liquid state: an overview","volume":"8","author":"Murasing","year":"2020","journal-title":"Int. J. Chem. Stud."},{"key":"B35","doi-asserted-by":"publisher","first-page":"4913","DOI":"10.1038\/srep04913","article-title":"Compounds from multilayer plastic bags cause reproductive failures in artificial insemination","volume":"4","author":"Nerin","year":"2014","journal-title":"Sci. Rep."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1171","DOI":"10.1095\/biolreprod38.5.1171","article-title":"Capacitation of bovine sperm by Heparin1","volume":"38","author":"Parrish","year":"1988","journal-title":"Biol. Reprod."},{"key":"B37","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1080\/19396368.2020.1754962","article-title":"Toxicity and antimicrobial effect of silver nanoparticles in swine sperms","volume":"66","author":"P\u00e9rez-Duran","year":"2020","journal-title":"Syst. Biol. Reprod. Med."},{"key":"B38","doi-asserted-by":"publisher","first-page":"292","DOI":"10.3390\/nano10020292","article-title":"Metal-based nanoparticles as antimicrobial agents: an overview","volume":"10","author":"S\u00e1nchez-L\u00f3pez","year":"2020","journal-title":"Nanomater. (Basel)"},{"key":"B39","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.freeradbiomed.2020.07.024","article-title":"Nanotechnology in reproduction: vitamin E nanoemulsions for reducing oxidative stress in sperm cells","volume":"160","author":"S\u00e1nchez-Rubio","year":"2020","journal-title":"Free Radic. Biol. Med."},{"key":"B40","doi-asserted-by":"publisher","first-page":"21","DOI":"10.15764\/NANO.2014.01003","article-title":"Cell membrane protective efficacy of ZnO nanoparticles","volume":"1","author":"Sanjay","year":"2014","journal-title":"Trans. Nanotechnol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"e20210021","DOI":"10.1590\/1984-3143-AR2021-0021","article-title":"Influence of antibiotics on bacterial load and sperm parameters during short-term preservation of collared peccary semen","volume":"18","author":"Santos","year":"2021","journal-title":"Anim. Reprod."},{"key":"B42","doi-asserted-by":"publisher","first-page":"78","DOI":"10.3390\/ani10010078","article-title":"Comparison between the effects of adding vitamins, trace elements, and nanoparticles to SHOTOR extender on the cryopreservation of dromedary camel epididymal spermatozoa","volume":"10","author":"Shahin","year":"2020","journal-title":"Anim. (Basel)"},{"key":"B43","doi-asserted-by":"publisher","first-page":"65","DOI":"10.22038\/nmj.2021.08.07","article-title":"Role of Rubia tinctorum in the synthesis of zinc oxide nanoparticles and apoptosis induction in breast cancer cell line","volume":"8","author":"Shamasi","year":"2021","journal-title":"Nanomed. J."},{"key":"B44","doi-asserted-by":"publisher","first-page":"4042","DOI":"10.3390\/ijms20164042","article-title":"Role of autophagy in zinc oxide nanoparticles-induced apoptosis of mouse LEYDIG cells","volume":"20","author":"Shen","year":"2019","journal-title":"Int. J. Mol. Sci."},{"key":"B45","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1093\/molehr\/3.3.203","article-title":"Biochemistry of the induction and prevention of lipoperoxidative damage in human spermatozoa","volume":"3","author":"Storey","year":"1997","journal-title":"Mol. Hum. Reprod."},{"key":"B46","doi-asserted-by":"publisher","first-page":"530","DOI":"10.1002\/j.1939-4640.1996.tb01830.x","article-title":"Lipid peroxidation and human sperm motility: protective role of vitamin E","volume":"17","author":"Suleiman","year":"1996","journal-title":"J. Androl."},{"key":"B47","doi-asserted-by":"publisher","first-page":"651","DOI":"10.3762\/bjnano.6.66","article-title":"Influence of gold, silver and gold\u2013silver alloy nanoparticles on germ cell function and embryo development","volume":"6","author":"Taylor","year":"2015","journal-title":"Beil. J. Nanotechnol."},{"key":"B48","doi-asserted-by":"publisher","first-page":"3583","DOI":"10.1016\/j.biomaterials.2008.05.015","article-title":"Size, charge and concentration dependent uptake of iron oxide particles by non-phagocytic cells","volume":"29","author":"Thorek","year":"2008","journal-title":"Biomaterials"},{"key":"B49","doi-asserted-by":"publisher","first-page":"1568","DOI":"10.3390\/nano10081568","article-title":"Iron oxide nanoparticles as an alternative to antibiotics additive on extended boar semen","volume":"10","author":"Tsakmakidis","year":"2020","journal-title":"Nanomater. (Basel)"},{"key":"B50","doi-asserted-by":"crossref","first-page":"20","DOI":"10.5455\/OVJ.2014.v4.i1.p20","article-title":"TRIXcell+, a new long-term boar semen extender containing whey protein with higher preservation capacity and litter size","volume":"4","author":"van den Berg","year":"2014","journal-title":"Open Vet. J."},{"key":"B51","doi-asserted-by":"publisher","first-page":"1987","DOI":"10.1021\/nn800631r","article-title":"Splendid one-dimensional nanostructures of zinc oxide: a new nanomaterial family for nanotechnology","volume":"2","author":"Wang","year":"2008","journal-title":"ACS Nano"}],"container-title":["Frontiers in Bioengineering and Biotechnology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fbioe.2024.1326143\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,23]],"date-time":"2024-02-23T05:02:02Z","timestamp":1708664522000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fbioe.2024.1326143\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,23]]},"references-count":51,"alternative-id":["10.3389\/fbioe.2024.1326143"],"URL":"https:\/\/doi.org\/10.3389\/fbioe.2024.1326143","relation":{},"ISSN":["2296-4185"],"issn-type":[{"value":"2296-4185","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,23]]},"article-number":"1326143"}}