Cryobiology最新文献

{"title":"Vitrification of porcine intact femoral condyle through different protocol lengths and loading strategies","authors":"Mohammadhamed Shahsavari ,&nbsp;Korinne M. Tirones ,&nbsp;Marggi A. Patel ,&nbsp;Janet A.W. Elliott ,&nbsp;Nadr M. Jomha","doi":"10.1016/j.cryobiol.2025.105215","DOIUrl":"10.1016/j.cryobiol.2025.105215","url":null,"abstract":"<div><div>Cryopreservation of articular cartilage (AC) allografts is a potential candidate to solve the shortage of fresh osteochondral allografts to treat large AC defects. This technique requires optimization to address issues related to the actual process of cryopreservation and the translation of the technology to tissue banks. In the present study, we compared different vitrification protocols consisting of multiple cryoprotective agent (CPA) loading timings and loading strategies to achieve successful vitrification-rewarming protocols for porcine intact femoral condyles. Porcine medial femoral condyles (n = 6) were used to test 5 vitrification protocols differing in lengths of time and the method of CPA loading. The effects of these protocols on post-warming normalized chondrocyte viability, metabolic activity, and tissue cracking frequency were documented. We demonstrated that all 5 CPA loading-vitrification protocols resulted in high normalized chondrocyte viability (&gt;88 % of Fresh control) post vitrification in porcine intact femoral condyles. Moreover, the Room Temperature Gel (RG) protocol showed high viability and metabolic activity of chondrocytes post-warming, and also decreased cracking frequency in AC after rewarming of vitrified intact femoral condyles. To facilitate translation of this technology to tissue banks, we showed that by shortening (total time of 15 min in Protocol RG) or combining our established Protocol 8 (430 min) with a hydrogel, including use of cryobags in liquid nitrogen vapour as the conventional storage method in tissue banks, we could introduce two reproducible protocols for clinical applications involving vitrified osteochondral tissue.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105215"},"PeriodicalIF":2.3,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143476328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"BGP-15 improves quality of goat sperm by mitigating oxidative stress during cryopreservation","authors":"Fuqin Liu ,&nbsp;Jianjun Dai ,&nbsp;Jun Gao ,&nbsp;Mengqian He ,&nbsp;Jiehuan Xu ,&nbsp;Caifeng Wu ,&nbsp;Shushan Zhang ,&nbsp;Xing Zhu ,&nbsp;Lingwei Sun","doi":"10.1016/j.cryobiol.2025.105232","DOIUrl":"10.1016/j.cryobiol.2025.105232","url":null,"abstract":"<div><div>The objective of this study was to evaluate the effect of the addition of BGP-15 ((Z)-N'-(2-hydroxy-3-(piperidin-1-yl) propoxy) nicotinimidamide) on goat sperm during cryopreservation. The semen from six rams was diluted with a soybean lecithin-based extender containing different doses of BGP-15 (0, 50, 100, 150, and 200 μM). After the freeze-thawing process, sperm characteristics, plasma membrane integrity and functionality, acrosomal status, mitochondria activity, apoptosis status, and gene expression of post-thawed ram spermatozoa were assessed. Compared to fresh spermatozoa, the ultrastructure of frozen spermatozoa was damaged, and frozen spermatozoa had a reduced number of duplex microtubules with partially asymmetric distributions, localized disappearance of the central microtubule, and ambiguous submicrotubule structures. Moreover, the 100 μM BGP-15 treatment group had the highest percentage of viability, plasma membrane and acrosome integrities, and mitochondrial activity of frozen-thawed spermatozoa when compared to the control (P &lt; 0.05). In the Computer-Assisted Semen Analyzer, sperm motility parameters were significantly increased in the BGP-15 treatment group against the control group (P &lt; 0.05), except wobble. Similarly, BGP-15 treatment increased the levels of T-AOC, SOD, CAT, and GHS-Px and decreased the level of ROS which compared to controls (P &lt; 0.05). Only the 100 μM BGP-15 treatment group had a higher MDA level than the control group (P &lt; 0.05). In all BGP-15 treatment groups, the mRNA expressions of the <em>ROMO1</em> gene were significantly reduced compared to controls (P &lt; 0.05), and the <em>mRNA</em> expressions of the <em>SMCP, MnSOD</em>, <em>and CuZnSOD</em> genes were significantly increased (P &lt; 0.05). While the difference in the <em>mRNA</em> expression of the <em>SMOX</em> gene between the 50 μM BGP-15 treatment group and the control group was not significant (P &gt; 0.05). Moreover, the apoptosis rate of freeze-thawed spermatozoa significantly decreased in the 100 μM BGP-15 treatment group compared to the control (P &lt; 0.05), while the MMP of freeze-thawed spermatozoa significantly enhanced in the 100 μM BGP-15 treatment group (P &lt; 0.05). In conclusion, BGP-15 enhances cryo-protective effects on goat spermatozoa, and 100 μM BGP-15 addition to the extender during cryopreservation is beneficial to the goat breeding industry.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105232"},"PeriodicalIF":2.3,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143620760","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of the effect of human testicular cell conditioned media on the in vitro development of follicles from cryopreserved human ovarian cortical pieces. A potential approach for fertility preservation for cancer patients","authors":"Mohammad Ali Khalili ,&nbsp;Behrouz Aflatoonian ,&nbsp;Mohammad Reza Mirzaei ,&nbsp;Mahin Izadi ,&nbsp;Mojgan Noroozi Karimabad ,&nbsp;Fatemeh Asadi ,&nbsp;Mahboubeh Vatanparast","doi":"10.1016/j.cryobiol.2025.105218","DOIUrl":"10.1016/j.cryobiol.2025.105218","url":null,"abstract":"<div><div>If in vitro culture conditions could be improved, in vitro growth of cryopreserved ovarian tissue and isolated follicles could become an alternative to ovarian tissue transplantation. This study evaluated two cryopreservation methods, slow cooling and vitrification, and two in vitro culture methods, to determine their effect on the in vitro growth of human ovarian follicles. After warming, the OT pieces (1 × 10 × 3 mm) were cultured in either routine culture medium (DMEM), or DMEM supplemented 50:50 with media previously used for human testicular cell culture (hTCCM) to serve as a source of growth factors. Several parameters were evaluated during culture including follicle recruitment, growth, morphology, diameter, hormone production, and gene expressions (PTEN, BMP-15, PDGF, GDF-9, and GAPDH). The follicular morphology and hormonal secretion were comparable for both cryopreservation methods and both culture methods (P &gt; 0.05). Follicle recruitment was accelerated in the vitrified group, in the presence of hTCCM (P &gt; 0.05). After 7 days of culture, the follicle diameter was greater in the slow/hTCCM compared to the vitrification DMEM group. Similarly only one gene expression profile, BMP-15, in the slow/hTCCM differed significantly from another treatment group (Vit DMEM). At the end of culture (over 21 days), three immature, low-quality oocytes were the achievement. In conclusion, a very quick and easy vitrification protocol gave comparable results to the slow cooling method. We also established that the laboriously prepared hTCCM supplement did not benefit the outcomes. Further work is needed before in vitro maturation of cryopreserved ovarian cortical pieces can be used clinically.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105218"},"PeriodicalIF":2.3,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143609661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect of cryopreservation on enamel microcracks – A μCT analysis using a deep learning algorithm","authors":"Noëmi M.C. De Roo ,&nbsp;Pierre Kibleur ,&nbsp;Liesbeth Temmerman ,&nbsp;Ronald M.H. Verbeeck ,&nbsp;Matthieu N. Boone ,&nbsp;Guy A.M. De Pauw","doi":"10.1016/j.cryobiol.2025.105207","DOIUrl":"10.1016/j.cryobiol.2025.105207","url":null,"abstract":"<div><div>To date, the effect of cryopreservation on microcracks in the dental enamel remains unclear. These enamel microcracks are very thin, at the limit of visibility and their segmentation is beyond the capabilities of traditional image analysis. The objective of the present study was to investigate the effect of cryopreservation on enamel microcracks with a μCT analysis using a deep learning algorithm.</div><div>A manual annotation was performed to construct a high-quality training and testing dataset. A 4-phase semantic segmentation U-Net architecture neural network was trained in Dragonfly (ORS systems, Canada) and then applied on a sample of 5 teeth before and after cryopreservation, enabling for the first time a direct evaluation of the formation and evolution of enamel cracks caused by cryopreservation.</div><div>Qualitatively, the segmentation results were very satisfying and cracks as thin as 2–3 voxels wide could be segmented automatically. All teeth presented enamel microcracks, without propagation into the dentin. Similar crack patterns were observed in all teeth and may be related to the use of forceps during extraction. In the post-cryopreservation scans the damage extended, and new smaller cracks appeared on the occlusal surface of the tooth. Quantitatively, the average crack/enamel ratio was 0.066 ± 0.021 % before cryopreservation, and 0.087 ± 0.018 % after cryopreservation.</div><div>The present study presents the first scalable yet precise method to quantify clinically relevant tooth damage after cryopreservation. As such, this method also opens the way to future in-depth studies on dental enamel in many dental fields.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105207"},"PeriodicalIF":2.3,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143390452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of long-term storage on the quality of cryopreserved sperm of the Pangasius nasutus (Bleeker, 1863)","authors":"Nurizzati Idris ,&nbsp;Donald Torsabo ,&nbsp;Muhammad Yazed Abduh ,&nbsp;Ambok Bolong Abol-Munafi ,&nbsp;Noordiyana Mat Noordin ,&nbsp;Ivan Chong Chu Koh","doi":"10.1016/j.cryobiol.2025.105219","DOIUrl":"10.1016/j.cryobiol.2025.105219","url":null,"abstract":"<div><div>In the present study, we investigated the effects of storage for 3, 6, 9 and 12 months on cryopreserved sperm of <em>P. nasutus</em>, in 10 % MeOH as a cryoprotectant with 90 % 0.9 % NaCl as an extender. Sperm quality on motility, fertilization, hatching, and deformity rates were investigated using 9-months cryopreserved sperm. Determination on the effects of different sperm to egg ratios also was evaluated using 1-year cryopreserved sperm. Post-thaw motility (PTM) of cryopreserved sperm within storage period was significantly lower compared to the initial motility of fresh sperm (51.67 ± 2.4 %). However, there was no significant change of PTM during 12 months of storage (3 months, 33.3 ± 2.33 %; 6 months, 32.0 ± 2.52; 9 months, 32.67 ± 1.76; 12 months, 33.33 ± 1.67). In addition, the motility duration was unaffected by storage as there was no significant difference compared to fresh samples. Besides that, there were no significant differences in motility, fertilization and deformity rates for fresh and 9-months cryopreserved sperm except for hatching rate. Samples stored for 1 year resulted in high fertilization across all sperm to egg ratios, showing no difference compared to fresh sperm. Cryopreserved sperm exhibited a significant higher hatching rate (9.24 ± 4.68 %) at the sperm to egg ratio of 300,000:1 compared to 150,000:1 (3.44 ± 1.7 %). However, no significant difference in deformity rates for fresh and cryopreserved at sperm to egg ratio of 300,000:1. The newly developed cryopreservation protocol using methanol successfully preserved sperm quality, maintaining fertilization rates comparable to fresh sperm. This highlights its potential application in sustainable aquaculture and conservation strategies for <em>Pangasius nasutus</em>.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105219"},"PeriodicalIF":2.3,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143593992","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Potential application of acid-form sophorolipids in cell cryopreservation","authors":"Thi Nhu Trang Nguyen,&nbsp;Yuki Saito,&nbsp;Motoki Tatsumi,&nbsp;Masashi Yamamoto,&nbsp;Yoshihiko Hirata","doi":"10.1016/j.cryobiol.2025.105228","DOIUrl":"10.1016/j.cryobiol.2025.105228","url":null,"abstract":"<div><div>In cryopreservation, which is an important process for the long-term storage and transport of cells, cryopreservation solutions containing cryoprotectants are usually used to protect cells from damage. However, most cryoprotectants have non-negligible cytotoxicity and side effects that greatly limit their applications in clinical use. This has led to an increased need for more biocompatible cryoprotectants, and interest in bioinspired cryoprotectants is increasing. In this study, we have discovered the potential of using natural acid-form sophorolipids (aSL) as a cryoprotectant, which suppresses ice formation and reduces osmotic stress. The solution containing aSL showed the smallest size of ice crystals compared to the solutions containing other cryoprotectants, such as dimethyl sulfoxide (Me2SO), glycerol (GL), ethylene glycol (EG), and propylene glycol (PG). By adding aSL to a hypertonic culture medium, cell viability was significantly improved. This finding suggests an opportunity to develop low-toxicity and efficient reagents for cell cryopreservation in the future.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105228"},"PeriodicalIF":2.3,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143593993","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of vitrification on in vitro developmental competence of rat testicular tissue","authors":"Mina Sharbatoghli ,&nbsp;Samira Hajiaghalou ,&nbsp;Nafiseh Sadat Deheshkar Gooneh Farahani ,&nbsp;Samaneh Aghajanpour ,&nbsp;Abdolhossein Shahverdi ,&nbsp;Mojtaba Rezazadeh Valojerdi ,&nbsp;Bita Ebrahimi","doi":"10.1016/j.cryobiol.2025.105213","DOIUrl":"10.1016/j.cryobiol.2025.105213","url":null,"abstract":"<div><div>Cryopreservation of testicular tissue has been proposed as a potential technique for preserving fertility in pre-pubertal boys with various malignancies. The present study aimed to compare the effects of two vitrification techniques—solid surface vitrification (SSV) and needle-immersed vitrification (NIV)—on the integrity, development, cell viability, and apoptosis of rat testicular tissue. Testes from 4-week-old Wistar rats underwent a two-step vitrification process. Tissue pieces were allocated to either the SSV or NIV group. Equilibration involved a solution containing 7.5 % dimethyl sulfoxide (DMSO) and 7.5 % ethylene glycol (EG), followed by a vitrification solution with 0.07 mol/L sucrose, 15 % DMSO, and 15 % EG. The optimal protocol was determined after vitrification using either the NIV or SSV technique. Samples from the control and selected vitrification (SSV) groups were cultured for 3 weeks. Tissue integrity, cell viability, apoptosis, and gene expression were evaluated using hematoxylin and eosin staining, trypan blue staining, annexin V-PI staining, and real-time PCR. Morphological changes were more pronounced in the NIV group compared to the SSV group (P &lt; 0.05). Although the percentage of viable cells did not significantly differ between the NIV and SSV groups, it was slightly higher in the SSV group. Thus, SSV was identified as the optimum vitrification method. Real-time PCR analysis revealed altered gene expression: spermatogonial-related genes (<em>Lrp4</em>, <em>Egr3</em>, <em>Nanos</em>, <em>Gfra1</em>, <em>C-kit</em>, and <em>Sohlh1</em>) were significantly decreased in the SSV group, while somatic-cells-related genes (<em>Gdnf</em>, <em>Csf1</em>, and <em>Fgf2</em>) were higher. Overall, SSV appears suitable for rat testis tissue vitrification, although it induces some molecular changes. Optimization of the culture medium is essential to support successful spermatogenesis.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105213"},"PeriodicalIF":2.3,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143425143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Long term stability of preservative-free and lyophilized PRGF eye drops stored at different temperature conditions: in vitro comparative study","authors":"Eduardo Anitua ,&nbsp;María de la Fuente ,&nbsp;Mohammad Hamdan Alkhraisat","doi":"10.1016/j.cryobiol.2025.105214","DOIUrl":"10.1016/j.cryobiol.2025.105214","url":null,"abstract":"<div><div>Long-term stability of blood-derived eye drops is required to adapt the use of this treatment to prolonged clinical treatments and more green pharmacy. This study has been designed to assess the long-term storage life of freeze-dried plasma rich in growth factors (PRGF) eye drops, maintaining their biological content and activity. Thus, blood from five healthy donors was extracted and centrifuged for obtaining PRGF. The obtained PRGF eye-drops after platelet activation were freeze-stored or were lyophilized and then stored for 18 and 24 months at room temperature (RT) or at + 5 °C. Growth factor content and proliferative potential of PRGF eye drops on primary human keratocytes (HK) was evaluated at each storage time and condition. All growth factors maintained their levels at each time and storage condition. No differences were observed on the proliferative activity of keratocytes after treatment with freeze-dried PRGF eye-drops stored at RT or +5 °C for 18 or 24 months in comparison with fresh samples. No microbial contamination was observed in any of the PRGF eye-drops. Accepting the limitations of this study, it is observed that freeze-dried PRGF eye drops retain both key growth factors and biological activity when stored at room temperature or +5 °C for up to 18–24 months.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105214"},"PeriodicalIF":2.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143432687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “Cryopreservation of porcine skin-derived stem cells using melatonin or trehalose maintains their ability to self-renew and differentiate” [Cryobiology 107 (2022) 23–34]","authors":"Jia-Dong Sun ,&nbsp;Yu Sun ,&nbsp;Tian Qiao ,&nbsp;Shu-Er Zhang ,&nbsp;Paul W. Dyce ,&nbsp;Yuan-Wei Geng ,&nbsp;Ping Wang ,&nbsp;Wei Ge ,&nbsp;Wei Shen ,&nbsp;Shun-Feng Cheng","doi":"10.1016/j.cryobiol.2025.105201","DOIUrl":"10.1016/j.cryobiol.2025.105201","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105201"},"PeriodicalIF":2.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of resveratrol on post-thaw motility, kinematics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa","authors":"Tarique Hussain ,&nbsp;Muhammad Hammad Fayyaz ,&nbsp;Amjad Hameed ,&nbsp;Syed Murtaz Hassan Andrabi ,&nbsp;Rehana Kausar ,&nbsp;Yasin Mubashir ,&nbsp;Iqra Batool ,&nbsp;Muhammad Shahzad ,&nbsp;Ali Dogan Omur","doi":"10.1016/j.cryobiol.2025.105202","DOIUrl":"10.1016/j.cryobiol.2025.105202","url":null,"abstract":"<div><div>Resveratrol is a polyphenol compound showing strong antioxidant properties. It is believed that semen cryopreservation causes significant sperm losses which eventually affects sperm quality. Improving antioxidant status of semen may reduce this damage and enhance sperm fertilizing potential. This study investigated the resveratrol treated freeze-thawing sperm motion, kinetics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa. Thirty-two ejaculates from 4 fertile Kamori bucks were processed in tris-citric-glyercol-egg yolk (TCG-EY) based with varying concentrations of resveratrol (0, 10, 20, 40 and 50 μM). Over 75 % sperm motility were pooled and frozen in liquid nitrogen. The results unveiled that adding resveratrol at 40 and 50 μM concentration significantly enhanced (<em>P</em> &lt; 0.05) total motility (progressive motility, rapid velocity, and average path velocity, straight line velocity, curvilinear velocity, amplitude of lateral head displacement, beat cross frequency, straightness and linearity in contrast with control and other groups. Supplementing resveratrol at 40 and 50 μM concentration significantly improved (<em>P</em> &lt; 0.05) functional plasma membrane integrity, acrosome integrity, whereas, all resveratrol groups had same significant (<em>P</em> &lt; 0.05) effect on DNA integrity in response to control group. The 40 and 50 μM resveratrol significantly promoted (P &lt; 0.05) superoxide dismutase (SOD), catalase (CAT), peroxidases (POD), and total antioxidant capacity (TAC) levels while significantly reducing (<em>P</em> &lt; 0.05) the total oxidant status (TOS) and malondialdehyde (MDA) contents as compared to control and other groups, respectively. The principal component analysis (PCA) exhibited samples were present in different clusters except two groups which had partially overlapped. Using hierarchical clustering analysis, two clusters were constructed showing the relationship within the treated groups. The results of spearman correlation coefficient showed that yellow color showed highly positive correlation while turquoise color exhibited highly negative association between sperm variables. Overall, the results concluded that resveratrol at 50 μM performed slightly better results than 40 μM in terms of improving sperm quality parameters.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105202"},"PeriodicalIF":2.3,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143001673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}