Cryobiology最新文献

{"title":"Nanophytosomal delivery of resveratrol as an effective strategy for enhancing frozen–thawed bovine sperm quality","authors":"Ali Ali El-Raghi ,&nbsp;Amer K. Mohammed ,&nbsp;Ekramy M. Elmorsy ,&nbsp;Mahmoud A.E. Hassan ,&nbsp;Walaa M. Essawi ,&nbsp;Suzan Shawky Abuelkasem Mohamed ,&nbsp;Abeer M.E. Hassan ,&nbsp;Soha A. Hassan","doi":"10.1016/j.cryobiol.2025.105574","DOIUrl":"10.1016/j.cryobiol.2025.105574","url":null,"abstract":"<div><div>The aim of this study was to assess the effects of supplementing semen extender with free resveratrol (FR) or resveratrol-loaded nano-phytosomes (RPNPs) on the quality, antioxidant capacity, enzymatic activities, and apoptotic genes of cryopreserved bovine sperm. Semen samples were collected from five proven fertility Holstein cattle bulls aged 4–6 years using the artificial vagina method. Pooled semen was cryopreserved in a Tris-based extender supplemented with free resveratrol (FR) at a concentration of 90 μg/mL and RPNPs at concentrations of 30 μg/mL (RPNPs30), 60 μg/mL (RPNPs60), and 90 μg/mL (RPNPs90). Cryopreserved semen without any additives served as the control. Supplementing the freezing medium with free resveratrol (FR) or RPNPs at 60 or 90 μg/mL significantly enhanced sperm progressive motility, viability, membrane integrity, and kinematic parameters (p &lt; 0.05). Additionally, RPNPs60 and RPNPs90 treated samples exhibited superior antioxidant activities (TAC, GPX, and SOD), reduced lipid peroxidation, and improved enzymatic activities in seminal plasma compared to the control group (p &lt; 0.05). The supplementation of FR or RPNPs significantly improved the transcription of apoptotic genes such as <em>caspase 3, Bcl2,</em> and <em>Bax</em> in sperm. Electron microscopy observations revealed that fortifying the freezing media with 60 or 90 μg of RPNPs maintained the integrities of the acrosome and plasma membrane, and preserved the ultrastructure integrity of cryopreserved bovine spermatozoa. The docking analysis revealed that the binding affinity with key sperm function biomarkers, such as CAT, SOD, GPX, Caspase, Bax, and BCL2, exhibited binding energies of −8.5, −7.9, −7.5, −10.4, −9.0, and −9.8 kcal/mol, respectively. The addition of 60–90 μg RPNPs to the bovine freezing extender improved post-thawed sperm quality.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105574"},"PeriodicalIF":2.1,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145877962","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":"Optimizing ovarian tissue preparation methods for vitrification: A two-part study on slicing methods and tissue size effects","authors":"Y. Zhang,&nbsp;K. Beilby,&nbsp;S. Catt,&nbsp;M. Pangestu","doi":"10.1016/j.cryobiol.2025.105572","DOIUrl":"10.1016/j.cryobiol.2025.105572","url":null,"abstract":"<div><div>Ovarian tissue cryopreservation by vitrification is well-established, but the effects of procedural differences in the preparatory steps, such as dissection method and tissue size, remain underexplored. This study compared the thickness of cortical tissue pieces and the normality of primordial follicles prepared with either a Stadie Riggs tissue slicer or a conventional scalpel. Part two of the study assessed the viability of follicles vitrified within either 0.5–1 × 5 × 5 mm (thickness × length × width) or 0.5–1 × 10 × 5 mm pieces of cortical tissue. In part one, the proportion of morphologically normal follicles and section thickness did not differ by method (<span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>309</mn></mrow></math></span> and <span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>986</mn></mrow></math></span>), although thickness varied by operator (<span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>028</mn></mrow></math></span>) without an interaction between method and operator. Upon visual H&amp;E analysis, ovarian pieces prepared with the Stadie Riggs devices were more uniform in thickness and the sectioned edge less jagged than those prepared with a conventional scalpel. In part two, follicles were assessed on matched H&amp;E, Ki67 and <span><math><mi>γ</mi></math></span>H2AX stained sections before and after four days of <em>in vitro</em> culture. Fresh tissue processed on day 0 yielded higher proportions of ‘good’ outcomes for morphology, proliferation, and DNA-damage compared to vitrified tissues (<span><math><mrow><mi>p</mi><mo>&lt;</mo><mn>0</mn><mo>.</mo><mn>001</mn></mrow></math></span>, <span><math><mrow><mi>p</mi><mo>&lt;</mo><mn>0</mn><mo>.</mo><mn>001</mn></mrow></math></span> and <span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>001</mn></mrow></math></span>, respectively). By day 4, group differences were not significant. Direct comparisons between vitrified 0.5–1 × 5 <span><math><mo>×</mo></math></span> 5 mm and 0.5–1 × 10 <span><math><mo>×</mo></math></span> 5 mm vitrified pieces did not show significant differences at either time point (all <span><math><mrow><mi>p</mi><mo>&gt;</mo><mn>0</mn><mo>.</mo><mn>05</mn></mrow></math></span>); pooled analyses yielded odds ratios <span><math><mo>∼</mo></math></span>1 with no heterogeneity. In summary, both preparation methods provide comparable quantitative outcomes, with operator standardization appearing more influential than device choice. For vitrified ovarian cortex, both tissue sizes are interchangeable, and fresh tissue is superior at baseline. Considering the smoother cuts observed, we recommend Stadie Riggs preparation with either 0.5–1 × 5 <span><math><mo>×</mo></math></span> 5 or 0.5–1 × 10 <span><math><mo>×</mo></math></span> 5 mm fragments.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105572"},"PeriodicalIF":2.1,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145877982","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":"A brief overview of the development of oocyte and embryo cryopreservation strategies with a focus on the roles of sugars","authors":"Krzysztof Papis","doi":"10.1016/j.cryobiol.2025.105577","DOIUrl":"10.1016/j.cryobiol.2025.105577","url":null,"abstract":"<div><div>In this brief review, I have attempted to summarize subjectively the history of disaccharide use in gamete and embryo cryopreservation, which began at least 90 years ago and has gradually become essential in many aspects of these procedures, such as the formulation of cryogenic solutions to aid dehydration prior to cryopreservation and the safe handling of cells after thawing/warming during the rehydration process. This is also, to a large extent, the history of the author's scientific activity in the field of freezing and vitrification of animal and human embryos and oocytes over the past 40 years.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105577"},"PeriodicalIF":2.1,"publicationDate":"2025-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145878001","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":"Influence of storage conditions on viability of hematopoietic stem cells and leukocyte subpopulations in fresh and cryopreserved umbilical cord blood samples","authors":"Vladimira Rimac ,&nbsp;Sanja Mazić ,&nbsp;Ines Bojanić","doi":"10.1016/j.cryobiol.2025.105576","DOIUrl":"10.1016/j.cryobiol.2025.105576","url":null,"abstract":"<div><div>Umbilical cord blood (UCB) units are cryopreserved for long-term storage, but it is still not fully investigated how cryopreservation and freezing affect the viability of different cell types. This prospective study evaluated the stability of fresh and cryopreserved UCB samples in various storage conditions using the 7-AAD/annexin V method. UCBs were collected “ex-utero” and processed according to the institutional standard operating procedures. In the stability study, fresh UCB buffy coat (BC) samples were stored at +4 °C and room temperature (RT) for up to 24 h, while thawed samples for up to one and 2 h. After a defined time period, cells were labelled again and analysed. Early apoptosis was most prevalent in CD34⁺ cells and least in T lymphocytes in both fresh and thawed samples. The highest post-thaw recovery was observed for T lymphocytes. The total CD19⁺ (P = 0.001) and CD16+/56+ (P = 0.004) cell counts were statistically significantly reduced when fresh samples were stored at RT. Total NK cell counts were also reduced when samples were stored at +4 °C (P = 0.036). In cryopreserved samples, there was statistically significant differences in total cell counts for all cell populations when samples were stored at RT, and under these storage conditions, early apoptosis of B and NK cells also occurred. The results showed different post-thaw recoveries of leukocyte subpopulations in the samples of cryopreserved UCB units. Cell exposure to the cryoprotective solution post-thaw does not affect total cell count or further development of apoptosis when UCB samples are stored at +4 °C for up to 2 h.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105576"},"PeriodicalIF":2.1,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145862249","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 viability of dermatophytes: a comparative evaluation of cryopreservation media","authors":"Rita de Cassia Santos da Silva ,&nbsp;Juliana Gomes de Souza Oliveira ,&nbsp;Juan Diego Ribeiro de Almeida ,&nbsp;Naira Sulany Oliveira de Sousa ,&nbsp;Érica Simplício de Souza ,&nbsp;Ana Cláudia Alves Cortez ,&nbsp;Flávia da Silva Fernandes ,&nbsp;Maurício Ogusku ,&nbsp;Hagen Frickmann ,&nbsp;João Vicente Braga de Souza","doi":"10.1016/j.cryobiol.2025.105575","DOIUrl":"10.1016/j.cryobiol.2025.105575","url":null,"abstract":"<div><div>Reliable long-term preservation of dermatophytes is essential for diagnostics, taxonomy, and experimental reproducibility, but simple and comparative protocols remain scarce. We evaluated viability and colony morphology of 59 clinical isolates of <em>Epidermophyton floccosum</em>, <em>Microsporum canis</em>, <em>M. gypseum</em>, <em>Trichophyton mentagrophytes</em>, <em>T. tonsurans</em> and <em>T. rubrum</em> after about 15 years of storage under five conditions: Sauton broth either with 3 % or 10 % (v/v) glycerol at −80 °C; Sabouraud dextrose broth (SDB) with 0 % or 10 % (v/v) glycerol at −80 °C; and water storage at room-temperature (Castellani method, 25 °C). Overall recovery was highest with Sauton broth at low glycerol (79.7 %), followed by Castellani (42.4 %), Sauton broth at high glycerol (35.6 %), Sabouraud broth at low glycerol (27.1 %) and Sabouraud broth at high glycerol (5.1 %). Species-specific patterns were evident: all <em>T. tonsurans</em> and <em>T. mentagrophytes</em> isolates remained viable in Sauton broth at low glycerol; <em>T. rubrum</em> showed good recovery in both Sauton (73.3 %) broth at low glycerol and with the Castellani approach (86.7 %); <em>M. gypseum</em> performed best with Sauton broth at low glycerol (75 %) and Sabouraud broth at low glycerol (62.5 %); and <em>M. canis</em> survived only with the Castellani method (80 %). <em>M. canis</em> cultures showed an altered phenotype characterized by macroconidia loss after long-term maintenance in water. These findings support storage in Sauton broth at low glycerol at −80 °C as a broadly effective option for long-term dermatophyte preservation. This study highlights the need for screening collections before storage so that more difficult species, such as <em>M. canis</em>, can be identified and stored by other techniques.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105575"},"PeriodicalIF":2.1,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145836658","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":"Cryopreservation decreased lipopolysaccharide-induced immune response of endometrial stromal cells via inhibiting the expression of TRIF","authors":"Cuiting Yang ,&nbsp;Yan Zhang ,&nbsp;Chun Wang ,&nbsp;Lingxi Zhu ,&nbsp;Ming Zhang","doi":"10.1016/j.cryobiol.2025.105567","DOIUrl":"10.1016/j.cryobiol.2025.105567","url":null,"abstract":"<div><div>In the study, we investigated the effect of cryopreservation on the immune response of endometrial stromal cells (ESCs) to lipopolysaccharide (LPS). We exposed both frozen-thawed and untreated ESCs to LPS and measured cytokine levels as well as genes associated with the LPS-TLR4-cytokine signaling pathway. Results revealed that LPS induced significant upregulation of <em>IL-1β</em>, <em>IL-6</em>, <em>IL-8</em>, and <em>TNF-α</em> in vitro cultured untreated cells compared to frozen-thawed ones. Furthermore, mRNA levels related to the TRIF pathway, including <em>TRAM</em>, <em>TRAF-6</em>, and <em>IRF-7</em>, <em>NF-κB</em>, and <em>JNK</em> were significantly upregulated in untreated ESCs after LPS-stimulation but not in frozen-thawed cells. Moreover, transcription levels of TRIF signaling-related genes were notably lower in frozen-thawed cells. Additionally, TRIM69 overexpression rescued the cryopreservation-induced suppression of immune response. Overall, our findings suggest that cryopreservation attenuates the LPS-induced immune response in ESCs by inhibiting the TRIF pathway.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105567"},"PeriodicalIF":2.1,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827118","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":"Temperature-metabolism relationships above and below the body fluid freezing point in the freeze-tolerant white worm, Enchytraeus albidus","authors":"Mathias Engell Dahl Holmstrup ,&nbsp;Johannes Overgaard ,&nbsp;Stine Slotsbo","doi":"10.1016/j.cryobiol.2025.105571","DOIUrl":"10.1016/j.cryobiol.2025.105571","url":null,"abstract":"<div><div><em>Enchytraeus albidus</em> is a freeze-tolerant oligochaete with potential for long-term cryopreservation storage as live feed in aquaculture. In this study, we measured the metabolic rate (MR) of <em>E. albidus</em> across a range of temperatures (+20 to −14 °C) in both unfrozen and frozen states using intermittent flow-through CO₂ respirometry. Our results demonstrate an exponential decrease in MR with declining temperature, and a markedly steeper decline below the freezing point of body fluids. Frozen worms exhibited a Q₁₀ of 24.1, in contrast to 2.2 in unfrozen worms, indicating significant metabolic depression associated with freezing. We argue that this depression is strongly correlated with intracellular osmolality, which increases due to freeze-induced dehydration. Theoretical calculations based on MR data suggest that glycogen stores could support frozen aerobic metabolism for up to 3–4 months at −14 °C, aligning with overwinter survival observed in Arctic populations. These findings indicate that <em>E. albidus</em> primarily relies on aerobic metabolism when frozen and highlight its suitability for long-term storage as live feed under moderate freezing conditions. Our results provide valuable physiological knowledge for optimizing industrial cryopreservation strategies to balance survival and shelf-life in freeze-tolerant invertebrates.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105571"},"PeriodicalIF":2.1,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827147","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":"Characterizing the cryobiological response of a mouse cardiac endothelial cell line to interrupted slow cooling (graded freezing)","authors":"Elham Ashrafi ,&nbsp;Janet A.W. Elliott","doi":"10.1016/j.cryobiol.2025.105563","DOIUrl":"10.1016/j.cryobiol.2025.105563","url":null,"abstract":"<div><div>Mouse cardiac endothelial cells play a critical role in cardiovascular research and disease; hence, high quality cryopreservation of this cell type is necessary to provide on-demand access to the cells, quality control, and consistent outcomes between different experiments and batches. However, the cryopreservation of cardiac endothelial cells has not been studied as much as other endothelial cells. The objective of this study was to characterize the cryobiological response of an immortalized line of mouse cardiac endothelial cells in suspension using an interrupted slow cooling approach (graded freezing) and to validate cryopreservation protocols that provide high post-thaw viability and function. Three different cryopreservation protocols were validated using different cryoprotectant solutions: <em>i</em>) 10 % dimethyl sulfoxide + 90 % fetal bovine serum, <em>ii</em>) 5 % dimethyl sulfoxide + 6 % hydroxyethyl starch in complete growth medium, and <em>iii</em>) 10 % glycerol in complete growth medium; all three protocols resulted in high post-thaw membrane integrity assessed using dual fluorescent dye Syto13/GelRed. Two functional assays: <em>i</em>) tube forming on Matrigel representing angiogenesis and <em>ii</em>) nitric oxide synthesis using DAF-FM (4-amino-5-methylamino-2′,7′-difluorofluorescein) fluorescent dye revealed that cryopreserved cells retained comparable post-thaw function to non-cryopreserved cells. This study provides practical insight into successful cryopreservation of mouse cardiac endothelial cells.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105563"},"PeriodicalIF":2.1,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145818483","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":"Glucose and maltose supplementation in the freezing medium of Creole rooster spermatozoa improves in vivo fertility","authors":"Elías Segarra Zenteno ,&nbsp;Liliana Morocho ,&nbsp;Karen Vásquez ,&nbsp;Mauricio Duma ,&nbsp;Diego A. Galarza","doi":"10.1016/j.cryobiol.2025.105573","DOIUrl":"10.1016/j.cryobiol.2025.105573","url":null,"abstract":"<div><div>This study investigated the effects of glucose-maltose supplementation in freezing medium on the kinematic, viability, and fertility of Creole rooster spermatozoa. Fifty ejaculates from 10 Creole roosters were obtained in 5 weekly sessions to create 10 heterologous pools. Each pool was extended with Lake-Ravie plus 6 % dimethylacetamide and allocated to: GM1 (0.64 % glucose + 0.33 % maltose), GM2 (0.72 % glucose + 0.17 % maltose), or non-supplemented control. After thawing, the control presented higher motility and beat-cross frequency than GM1 and GM2 (P &lt; 0.05). Viability was greater in GM1 and control than in GM2 (P &lt; 0.05). Crucially, the fertility after artificial insemination of hens with frozen-thawed sperm from the GM1 group was higher (P &lt; 0.05) compared to the non-supplemented group. In conclusion, glucose-maltose supplementation to the freezing medium can improve the viability and fertility of frozen-thawed Creole rooster semen, supporting its use in genetic conservation and breeding programs.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105573"},"PeriodicalIF":2.1,"publicationDate":"2025-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786511","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":"Cryoprotectants and dynamics of exclusion zone water","authors":"Alexis Theodorou,&nbsp;Karel Pomeisl,&nbsp;Jan Pokorný,&nbsp;Irena Kratochvílová","doi":"10.1016/j.cryobiol.2025.105566","DOIUrl":"10.1016/j.cryobiol.2025.105566","url":null,"abstract":"<div><div>Hydrophilic membranes are integral components in a wide range of applications, particularly in biological systems, where their negatively charged surfaces resemble those of cell membranes and influence processes such as cryopreservation. This study investigates the relationship between the Exclusion Zone (EZ) near a Nafion membrane and its response to thermal changes. In this context, the EZ denotes an extended interfacial water region adjacent to hydrophilic surface, characterized by physicochemical properties differing from those of bulk water. The present work investigates how cryoprotectants such as polyethylene glycols (PEGs) and trehalose affect the EZ near Nafion surfaces across a range of temperatures, aiming to identify potential links between interfacial hydration and cryoprotective efficiency. Our observations showed that PEG and trehalose exhibit distinct effects on the EZ behavior, with trehalose showing milder suppression of the EZ size compared to PEG at the same concentration. Fluorescent tracking of dansyl-labeled cryoprotectants demonstrated their distribution near the EZ, revealing differential long-range interactions of interfacial water with PEG and trehalose. Herein, we provide the first evidence linking cryoprotectant activity to the EZ behavior, suggesting a role in interfacial water stabilization under cooling, with implications for cryopreservation and membrane-associated processes.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105566"},"PeriodicalIF":2.1,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773763","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}