Cryobiology最新文献_第3页

Influence of storage conditions on viability of hematopoietic stem cells and leukocyte subpopulations in fresh and cryopreserved umbilical cord blood samples 储存条件对新鲜和冷冻脐带血样本中造血干细胞和白细胞亚群活力的影响。

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-29 DOI: 10.1016/j.cryobiol.2025.105576

Vladimira Rimac , Sanja Mazić , Ines Bojanić

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.

脐带血（UCB）单位被冷冻保存以进行长期储存，但冷冻保存和冷冻如何影响不同细胞类型的活力仍未得到充分研究。本前瞻性研究使用7-AAD/膜联蛋白V法评估新鲜和冷冻UCB样品在不同储存条件下的稳定性。ucb是在“子宫外”收集的，并根据机构的标准操作程序进行处理。在稳定性研究中，新鲜的UCB黄外套（BC）样品在+4°C和室温（RT）下保存24小时，而解冻的样品则保存1和2小时。在规定的时间段后，再次标记细胞并进行分析。在新鲜和解冻样品中，早期凋亡在CD34+细胞中最普遍，在T淋巴细胞中最少。T淋巴细胞解冻后恢复最高。新鲜样品在室温下保存时，CD19+细胞总数（P = 0.001）和CD16+/56+细胞总数（P = 0.004）显著减少，NK细胞总数在+4℃下保存时也显著减少（P = 0.036）。在低温保存的样品中，当样品在室温下保存时，所有细胞群的总细胞计数有统计学意义的差异，并且在这种保存条件下，B细胞和NK细胞也发生了早期凋亡。结果显示，在冷冻保存的UCB单位样品中，白细胞亚群的解冻后恢复不同。当UCB样品在+4°C下保存长达2小时时，解冻后细胞暴露于冷冻保护液中不影响细胞总数或细胞凋亡的进一步发展。

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引用次数: 0

Long-term viability of dermatophytes: a comparative evaluation of cryopreservation media 皮肤植物的长期生存能力：低温保存介质的比较评价

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-25 DOI: 10.1016/j.cryobiol.2025.105575

Rita de Cassia Santos da Silva , Juliana Gomes de Souza Oliveira , Juan Diego Ribeiro de Almeida , Naira Sulany Oliveira de Sousa , Érica Simplício de Souza , Ana Cláudia Alves Cortez , Flávia da Silva Fernandes , Maurício Ogusku , Hagen Frickmann , João Vicente Braga de Souza

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 Epidermophyton floccosum, Microsporum canis, M. gypseum, Trichophyton mentagrophytes, T. tonsurans and T. rubrum 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 T. tonsurans and T. mentagrophytes isolates remained viable in Sauton broth at low glycerol; T. rubrum showed good recovery in both Sauton (73.3 %) broth at low glycerol and with the Castellani approach (86.7 %); M. gypseum performed best with Sauton broth at low glycerol (75 %) and Sabouraud broth at low glycerol (62.5 %); and M. canis survived only with the Castellani method (80 %). M. canis 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 M. canis, can be identified and stored by other techniques.

可靠的长期保存皮肤真菌对诊断、分类和实验可重复性至关重要，但简单和可比较的方案仍然很少。在五种条件下，对59株临床分离的絮状表皮菌、犬小孢子菌、gypseum、mentagrophytes、T. tonsurans和T. rubrum在- 80°C下添加3%或10% （v/v）甘油的Sauton汤中保存约15年后的活力和菌落形态进行了评估；在- 80℃下，加入0%或10% （v/v）甘油的Sabouraud葡萄糖肉汤（SDB）；在室温下储存水（Castellani法，25°C）。低甘油索顿肉汤的总回收率最高（79.7%），其次是Castellani肉汤（42.4%）、高甘油索顿肉汤（35.6%）、低甘油索顿肉汤（27.1%）和高甘油索顿肉汤（5.1%）。物种特异性模式明显：所有的T. tonsurans和T. mentagrophytes分离株在低甘油的Sauton肉汤中仍能存活；在低甘油的Sauton肉汤和Castellani肉汤中，红毛霉的回收率均较好（73.3%）；在低甘油（75%）的Sauton肉汤和低甘油（62.5%）的Sabouraud肉汤中，M. gypseum表现最好；犬支原体仅用Castellani法存活（80%）。犬支原体培养物在水中长期维持后表现出以大分生孢子丢失为特征的表型改变。这些发现支持在- 80°C低甘油的Sauton肉汤中作为长期保存皮肤真菌的广泛有效的选择。这项研究强调了在储存前对收集物进行筛选的必要性，以便通过其他技术识别和储存更困难的物种，如犬支原体。

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引用次数: 0

Cryopreservation decreased lipopolysaccharide-induced immune response of endometrial stromal cells via inhibiting the expression of TRIF 低温保存通过抑制TRIF的表达来降低脂多糖诱导的子宫内膜基质细胞免疫应答。

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-23 DOI: 10.1016/j.cryobiol.2025.105567

Cuiting Yang , Yan Zhang , Chun Wang , Lingxi Zhu , Ming Zhang

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 IL-1β, IL-6, IL-8, and TNF-α in vitro cultured untreated cells compared to frozen-thawed ones. Furthermore, mRNA levels related to the TRIF pathway, including TRAM, TRAF-6, and IRF-7, NF-κB, and JNK 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.

在这项研究中，我们研究了低温保存对子宫内膜基质细胞（ESCs）对脂多糖（LPS）免疫反应的影响。我们将冷冻解冻和未经处理的ESCs暴露于LPS中，并测量细胞因子水平以及与LPS- tlr4细胞因子信号通路相关的基因。结果显示，与冻融细胞相比，LPS诱导体外培养的未处理细胞IL-1β、IL-6、IL-8和TNF-α显著上调。此外，与TRIF通路相关的mRNA水平，包括TRAM、trf -6、IRF-7、NF-κB和JNK，在lps刺激后未处理的ESCs中显著上调，而在冻解冻细胞中则没有上调。此外，TRIF信号相关基因的转录水平在冻融细胞中明显降低。此外，TRIM69过表达挽救了低温保存诱导的免疫反应抑制。总的来说，我们的研究结果表明，低温保存通过抑制TRIF途径减弱了内皮细胞中lps诱导的免疫反应。

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引用次数: 0

Temperature-metabolism relationships above and below the body fluid freezing point in the freeze-tolerant white worm, Enchytraeus albidus 耐冻性白虫在体液冰点上下的温度代谢关系。

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-23 DOI: 10.1016/j.cryobiol.2025.105571

Mathias Engell Dahl Holmstrup , Johannes Overgaard , Stine Slotsbo

Enchytraeus albidus 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 E. albidus 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 E. albidus 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.

斑叶藻是一种耐寒寡毛藻，具有作为水产养殖活饲料长期冷冻保存的潜力。在这项研究中，我们使用间歇式CO2呼吸测量法测量了在非冷冻和冷冻状态下（+20至-14°C）的代谢率（MR）。我们的结果表明，随着温度的下降，MR呈指数下降，体液冰点以下的下降幅度明显更大。冷冻蠕虫的Q10为24.1，而未冷冻蠕虫的Q10为2.2，这表明与冷冻相关的代谢显著降低。我们认为这种抑制与细胞内渗透压密切相关，而细胞内渗透压由于冷冻引起的脱水而增加。基于磁共振数据的理论计算表明，糖原储存可以支持在-14°C下长达3-4个月的冷冻有氧代谢，与北极种群的越冬存活率一致。这些研究结果表明，在冷冻条件下，黄颡鱼主要依赖于有氧代谢，并强调其适合在中等冷冻条件下作为活饲料长期储存。我们的研究结果为优化工业冷冻保存策略以平衡耐寒无脊椎动物的生存和货架期提供了有价值的生理学知识。

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引用次数: 0

Characterizing the cryobiological response of a mouse cardiac endothelial cell line to interrupted slow cooling (graded freezing) 表征小鼠心脏内皮细胞系对中断缓慢冷却（分级冷冻）的低温生物学反应。

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-22 DOI: 10.1016/j.cryobiol.2025.105563

Elham Ashrafi , Janet A.W. Elliott

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: i) 10 % dimethyl sulfoxide + 90 % fetal bovine serum, ii) 5 % dimethyl sulfoxide + 6 % hydroxyethyl starch in complete growth medium, and iii) 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: i) tube forming on Matrigel representing angiogenesis and ii) 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.

小鼠心脏内皮细胞在心血管研究和疾病中发挥关键作用因此，这种细胞类型的高质量冷冻保存是必要的，以提供按需访问细胞，质量控制和不同实验和批次之间一致的结果。然而，心脏内皮细胞的低温保存还没有像其他内皮细胞那样得到广泛的研究。本研究的目的是表征永生化小鼠心脏内皮细胞系在暂停中使用中断缓慢冷却方法（分级冷冻）的低温生物学反应，并验证提供高解冻后活力和功能的低温保存方案。使用不同的冷冻保护剂溶液验证了三种不同的冷冻保存方案：i) 10%二甲亚砜+ 90%胎牛血清，ii) 5%二甲亚砜+ 6%羟乙基淀粉在完全生长培养基中，iii) 10%甘油在完全生长培养基中；使用双荧光染料Syto13/GelRed对这三种方案的解冻后膜完整性进行了评估。两项功能分析：i)在基质上形成管，代表血管生成；ii)使用DAF-FM（4-氨基-5-甲氨基-2',7'-二氟荧光素）荧光染料合成一氧化氮，结果显示，冷冻保存的细胞与非冷冻保存的细胞保留了相当的解冻后功能。本研究为成功低温保存小鼠心脏内皮细胞提供了实用的见解。

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引用次数: 0

Glucose and maltose supplementation in the freezing medium of Creole rooster spermatozoa improves in vivo fertility 在克里奥尔公鸡精子冷冻培养基中添加葡萄糖和麦芽糖可提高体内育性

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-19 DOI: 10.1016/j.cryobiol.2025.105573

Elías Segarra Zenteno , Liliana Morocho , Karen Vásquez , Mauricio Duma , Diego A. Galarza

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 < 0.05). Viability was greater in GM1 and control than in GM2 (P < 0.05). Crucially, the fertility after artificial insemination of hens with frozen-thawed sperm from the GM1 group was higher (P < 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.

本研究研究了在冷冻培养基中添加葡萄糖-麦芽糖对克里奥尔公鸡精子运动、活力和生育能力的影响。从10只克里奥尔公鸡中获得50次射精，每周5次，形成10个异源池。每个池添加Lake-Ravie + 6%二甲基乙酰胺，并分配为：GM1（0.64%葡萄糖+ 0.33%麦芽糖），GM2（0.72%葡萄糖+ 0.17%麦芽糖）或未添加对照。解冻后，对照比GM1和GM2表现出更高的运动性和热交叉频率（P < 0.05）。GM1和对照的存活率均高于GM2 （P < 0.05）。重要的是，与未添加GM1组相比，GM1冻融精子组母鸡人工授精后的受精率更高（P < 0.05）。综上所述，在冷冻培养基中添加葡萄糖-麦芽糖可以提高冷冻解冻的克里奥尔公鸡精液的活力和育性，支持其在遗传保护和育种计划中的应用。

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引用次数: 0

Cryoprotectants and dynamics of exclusion zone water 低温保护剂和禁区水动力学。

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-16 DOI: 10.1016/j.cryobiol.2025.105566

Alexis Theodorou, Karel Pomeisl, Jan Pokorný, Irena Kratochvílová

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.

亲水膜是广泛应用中不可或缺的组成部分，特别是在生物系统中，其带负电荷的表面类似于细胞膜，并影响冷冻保存等过程。本研究探讨了Nafion膜附近的隔离区（EZ）与其对热变化的响应之间的关系。在这种情况下，EZ表示靠近亲水性表面的扩展界面水区域，其物理化学性质与散装水不同。目前的工作研究了低温保护剂如聚乙二醇（peg）和海藻糖如何在一定温度范围内影响Nafion表面附近的EZ，旨在确定界面水化和低温保护效率之间的潜在联系。我们的观察表明，聚乙二醇和海藻糖对EZ行为有明显的影响，在相同浓度下，海藻糖对EZ大小的抑制比聚乙二醇更温和。丹酚标记的冷冻保护剂的荧光跟踪显示它们分布在EZ附近，揭示了界面水与聚乙二醇和海藻糖的不同远程相互作用。在此，我们提供了将冷冻保护剂活性与EZ行为联系起来的第一个证据，表明在冷却下界面水稳定中起作用，这对低温保存和膜相关过程具有重要意义。

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引用次数: 0

Extracellular vesicles as emerging bioactive modulators in sperm cryopreservation: Mechanisms and therapeutic potentials 细胞外囊泡作为精子冷冻保存中新兴的生物活性调节剂：机制和治疗潜力

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-13 DOI: 10.1016/j.cryobiol.2025.105568

Merve Deniz Tanrıkulu , Mesut Çevik , Mustafa Numan Bucak , Alper Koçyiğit

Sperm cryopreservation is a cornerstone of assisted reproduction and genetic conservation, yet the freeze-thaw process induces substantial structural and functional damage to spermatozoa. Recent progress in the field has positioned extracellular vesicles (EVs) as potential therapeutic tools, primarily because of their capacity to transport functional biomolecules that help counteract the cellular damage caused by cryopreservation. We then examine the types of cellular and molecular damages encountered during sperm cryopreservation and synthesize current evidence on the protective effects of EVs in this context. Particular attention is given to the mechanisms by which EVs may preserve sperm integrity, including membrane stabilization, antioxidant delivery, and modulation of signaling pathways. Despite encouraging findings, challenges remain, including the standardization of EVs isolation methods, optimal dosing, and delivery strategies. We also discuss the need for mechanism-of-action studies and multi-omics approaches to better understand EV-sperm interactions. Finally, future directions are outlined, including EVs engineering and clinical translation. Overall, EVs emerge as promising modulators in cryobiology, with potential to improve sperm preservation outcomes and fertility restoration strategies. This review offers an in-depth examination of EVs, covering the processes involved in their formation, various strategies employed for their isolation, and the techniques used to assess their properties. It also highlights their sources and functional significance in the male reproductive system.

精子冷冻保存是辅助生殖和遗传保护的基础，但冷冻解冻过程会对精子造成严重的结构和功能损伤。该领域的最新进展将细胞外囊泡（EVs）定位为潜在的治疗工具，主要是因为它们具有运输功能性生物分子的能力，有助于抵消低温保存引起的细胞损伤。然后，我们研究了精子冷冻保存过程中遇到的细胞和分子损伤类型，并综合了ev在这种情况下的保护作用的现有证据。特别关注ev可能保持精子完整性的机制，包括膜稳定、抗氧化传递和信号通路的调节。尽管有令人鼓舞的发现，但挑战依然存在，包括电动汽车隔离方法的标准化、最佳给药和给药策略。我们还讨论了作用机制研究和多组学方法的必要性，以更好地了解ev -精子相互作用。最后，展望了未来的发展方向，包括电动汽车工程和临床翻译。总的来说，ev在低温生物学中成为有前途的调节剂，具有改善精子保存结果和生育恢复策略的潜力。本文综述了ev的深入研究，包括其形成过程、分离ev的各种策略以及用于评估ev性质的技术。它还强调了它们在男性生殖系统中的来源和功能意义。

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引用次数: 0

Coaxial temperature controlled cryoprinting: A biomimetic technology inspired by the freezing survival mechanisms of the frog Rana sylvatica 同轴温度控制冷印：一种仿生技术，灵感来自蛙蛙的冷冻生存机制

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-13 DOI: 10.1016/j.cryobiol.2025.105569

Leo Lou , Linnea Warburton , Cristina Bilbao-Sainz , Nicholas Jean , Boris Rubinsky

Inspired by the freeze-survival mechanism of the frog Rana sylvatica, we developed a co-axial, self-crosslinking approach to be used when fabricating frozen scaffolds. Soft biomaterial can be fabricated into scaffolds using methods such as Temperature Controlled Cryoprinting (TCC), which uses a freezing plate to freeze the biomaterial as it is extruded. However, it remains a challenge to uniformly crosslink the scaffolds during thawing without them losing their shape. In this paper, we use coaxial printing to generate locally variable phase-transition compositions, mimicking the frog's glucose distribution during freezing, and electrical Joule heating to simulate metabolic heat production. Unlike previous methods, in our novel approach the crosslinker is located already within the frozen scaffold, and there is no need to submerge the scaffold in a bath. Both mathematical modeling and experimental validation confirmed the concept. A finite thawing-diffusion-crosslinking model was developed to analyze parameters such as temperature, thawing interface, concentration, and crosslinking interface. Experimental evaluations, including mechanical testing, heating test, swelling performance and electron microscope, demonstrated that coaxial self-crosslinking enables controlled thawing and crosslinking of frozen scaffolds. Results demonstrated that coaxial TCC self-crosslinking scaffolds are feasible, allowing for a more controlled thawing and crosslinking process in cryoprinted scaffolds.

受到蛙蛙（Rana sylvatica）冷冻存活机制的启发，我们开发了一种同轴、自交联的方法，用于制造冷冻支架。柔软的生物材料可以通过温度控制冷冻打印（TCC）等方法制成支架，TCC使用冷冻板在挤压生物材料时将其冷冻。然而，如何在解冻过程中使支架均匀交联而不失去其形状仍然是一个挑战。在本文中，我们使用同轴打印来生成局部可变的相变成分，模拟青蛙在冷冻过程中的葡萄糖分布，并使用电焦耳加热来模拟代谢热的产生。与以前的方法不同，在我们的新方法中，交联剂已经位于冷冻支架内，并且不需要将支架浸入浴缸中。数学模型和实验验证都证实了这一概念。建立了有限融化-扩散-交联模型，分析了温度、融化界面、浓度和交联界面等参数。包括力学测试、加热测试、膨胀性能和电子显微镜在内的实验评估表明，同轴自交联可以实现冷冻支架的可控解冻和交联。结果表明，同轴TCC自交联支架是可行的，允许在冻印支架中更可控的解冻和交联过程。

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引用次数: 0

Modulation of inflammation of lipid metabolism by sesamin, forskolin, α-linolenic acid, and their combination in a zebrafish model of cryo-injury and high cholesterol diet-induced myocardial infarction, followed by atherosclerosis 芝麻素、福斯克林、α-亚麻酸及其组合在低温损伤和高胆固醇饮食诱导的心肌梗死后动脉粥样硬化斑马鱼模型中的脂质代谢炎症调节作用

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-12 DOI: 10.1016/j.cryobiol.2025.105565

Abu Safana Biswas, Kamsagara Linganna Krishna, Ganavi Bethanagere Ramesha, Pooja Gandharvachari Achar

Cryo-injury has been a standard model used to simulate myocardial infarction (MI), which may induce reproducible myocardial necrosis under controlled conditions. In a Zebrafish (ZF) model of cryoinjury and a high-cholesterol diet (HCD), the study evaluates the preventive effects of sesamin, forskolin, and α-linolenic acid, either alone or in combination, against myocardial infarction with atherosclerosis. Various physiological, biochemical, and histological endpoints were used to evaluate the effectiveness of the treatment. A notable recovery of cardiac activity (as measured by ECG) and body weight regulation was observed when comparing the treated groups with the cryo-injured control. Decreased TNF-α and iNOS levels demonstrated effective molecular control of cryoinjury-induced inflammation. An improved lipid balance was also observed through lipid profiling of cardiac tissue and serum, which revealed significant increases in HDL levels and decreases in triglycerides, total cholesterol, LDL, and VLDL. The treatment with a combination of test compounds especially shows good efficacy towards myocardial infarction and the myocardial structure of ZF, followed by atherosclerosis. In our study, the combination effect of sesamin, forskolin, and α-linolenic acid has significant potential for regulating lipid levels, inflammatory markers, and myocardial structure integrity. It can be concluded that the combination treatment of the sesamin, forskolin, and α-linolenic acid may protect the cardiovascular system, and this combination can be used for further preclinical study with a rodent model for cardiovascular disease.

低温损伤已成为模拟心肌梗死（MI）的标准模型，在可控条件下可诱发可重复性心肌坏死。在冷冻损伤和高胆固醇饮食（HCD）的斑马鱼（ZF）模型中，研究评估了芝麻素、福斯克林和α-亚麻酸单独或联合使用对心肌梗死合并动脉粥样硬化的预防作用。使用各种生理、生化和组织学终点来评估治疗的有效性。与低温损伤对照组相比，治疗组心脏活动（ECG测量）和体重调节明显恢复。TNF-α和iNOS水平的降低显示了低温损伤诱导炎症的有效分子控制。通过心脏组织和血清的脂质分析也观察到脂质平衡的改善，这表明HDL水平显著增加，甘油三酯、总胆固醇、LDL和VLDL水平降低。试验性化合物联合治疗尤其对心肌梗死和ZF心肌结构有较好的疗效，其次是动脉粥样硬化。在我们的研究中，芝麻素、福斯克林和α-亚麻酸的联合作用在调节血脂水平、炎症标志物和心肌结构完整性方面具有显著的潜力。由此可见，芝麻素、福斯克林和α-亚麻酸联合用药对心血管系统有一定的保护作用，该联合用药可用于啮齿动物心血管疾病模型的临床前研究。

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