{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T04:07:22Z","timestamp":1791518842781,"version":"4.3.4"},"reference-count":48,"publisher":"SAGE Publications","issue":"4","license":[{"start":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T00:00:00Z","timestamp":1722470400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Biopreservation and Biobanking"],"published-print":{"date-parts":[[2024,8]]},"abstract":"<jats:p>\n                    Ozone has been used as a therapy tool in medical science for conditions such as ulcers, peritonitis, wounds, and mostly joint problems. Ozone therapy strengthens the resistance to infections by kick-starting antioxidant, anti-inflammatory, and immune modulation systems. Ozone creates a defensive response against oxidative stress in membranes and protects metabolism against reactive oxygen species (ROS). Sperm membranes are one of ROS's main targets; therefore, the cells' cryopreservation process requires more defensive elements for better results. This study aimed to investigate the protective effect of nano-ozone solution (NOS) on ram sperm cryopreservation and the influence of the process on various sperm parameters for post-thaw (0 hour) and postincubation (6 hours) time points. Samples were collected from six Merino rams in the breeding season by electroejaculation five times at 3-day intervals. The study was conducted by cryopreservation of the samples using a tris citric acid\u2013egg yolk-based extender. The samples were subjected to freezing in control and NOS (0.5, 1, and 2\u2009\u03bcg\/mL nano-ozone supplemented). Post-thaw motility, hypo-osmotic swelling test, acrosome (fluorescein isothiocyanate-conjugated Pisum sativum agglutinin [PSA-FITC]), and DNA integrities (terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL]) were evaluated with a phase-contrast microscope. Mitochondrial membrane potential (MMP) assessments were conducted by JC1-PI dual staining with a flow cytometer. Malondialdehyde and glutathione (GSH) levels were measured by a spectrophotometer. Sperm kinematics were investigated by a computer-assisted sperm analyzer (CASA) at the post-thaw time point. Compared with the control, relatively low doses of NOS (0.5 and 1\u2009\u03bcg\/mL) yielded better results in many parameters (motility, membrane and acrosomal integrities, MMP, various sperm kinematics, and GSH levels) (\n                    <jats:italic toggle=\"yes\">p<\/jats:italic>\n                    \u2009&lt;\u20090.05). The addition of low ozone doses to cryopreservation extenders improved the results compared with the control group at post-thaw and postincubation time points. Despite the valuable potential of nano-ozone supplementation in ram sperm cryopreservation, this subject requires further investigations with fertility trials soon.\n                  <\/jats:p>","DOI":"10.1089\/bio.2023.0073","type":"journal-article","created":{"date-parts":[[2024,1,24]],"date-time":"2024-01-24T11:33:11Z","timestamp":1706095991000},"page":"312-320","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":5,"title":["Evaluation of Cryopreserved Ram Sperm with Nano-Ozone Solution and Post-Thaw Life Span by Flow Cytometric Analysis"],"prefix":"10.1177","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4033-9749","authenticated-orcid":false,"given":"Mehmed Berk","family":"Toker","sequence":"first","affiliation":[{"name":"Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Bursa Uludag University, Bursa, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmet \u00dcmit","family":"Sabanc\u0131","sequence":"additional","affiliation":[{"name":"Bursa \u00c7ekirge State Hospital, Orthopedics and Traumatology Clinic, Bursa, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00fclcan","family":"Avc\u0131","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyon, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2975-2594","authenticated-orcid":false,"given":"Ahmet","family":"Aktar","sequence":"additional","affiliation":[{"name":"Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Bursa Uludag University, Bursa, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bar\u0131\u015f","family":"Denk","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyon, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7017-5485","authenticated-orcid":false,"given":"\u00d6zge","family":"Bari","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Bursa Uludag University, Bursa, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G\u00f6zde Rabia","family":"\u00d6zalp","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Bursa Uludag University, Bursa, T\u00fcrkiye."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2024,8,19]]},"reference":[{"key":"e_1_3_5_2_2","first-page":"1","article-title":"Desmosterol incorporation into ram sperm membrane before cryopreservation improves in vitro and in vivo fertility","volume":"9","author":"Carro M de las M","year":"2021","unstructured":"Carro M de las M, , Ram\u00edrez-Vasquez RRA, , Pe\u00f1alva DA, et al. Desmosterol incorporation into ram sperm membrane before cryopreservation improves in vitro and in vivo fertility. Front Cell Dev Biol, 2021; 9(June):1\u201314; doi: 10.3389\/fcell.2021.660165","journal-title":"Front Cell Dev Biol"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.3390\/ani12101311"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.3390\/ani12121492"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.12681\/jhvms.18014"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2020.183357"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2018.04.001"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2020.08.007"},{"issue":"3","key":"e_1_3_5_9_2","first-page":"39","article-title":"Effects of antioxidants on semen freezing","volume":"1","author":"Bucak MN","year":"2015","unstructured":"Bucak MN, , G\u00fcng\u00f6r \u015e, Sari\u00f6zkan S, et al. 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