1)上海理工大学健康科学与工程学院,生物系统热科学研究所,上海 200093;2)上海交通大学附属第一人民医院临床泌尿外科中心/泌尿男科,上海 200080
1)Institute of Biothermal Technology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China;2)Department of Andrology, Clinical Center of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China
目的 针对青春期前及急需治疗的恶性肿瘤患者,卵巢组织的冷冻和移植是更合适的生育力保存方法。目前临床上通常采用高浓度的低温保护剂(cryoprotectants,CPAs)进行卵巢组织冷冻并采用水浴进行复温,这会导致卵巢组织受到不同程度的毒性损伤和反玻璃化损伤。因此,本文基于海藻酸钠水凝胶包封及磁热复温技术,提出了优化卵巢组织玻璃化保存的方法。方法 首先研究了海藻酸钠浓度、水凝胶包封与保护剂加载的顺序对包封卵巢组织玻璃化冷冻效果的影响,并验证了海藻酸钠水凝胶包封对CPAs浓度的降低作用。其次,搭建了水浴结合磁热复温的平台,在不同磁纳米粒子浓度及磁场强度条件下对卵巢组织进行复温,评估了复温后卵泡存活率、抗氧化能力及卵巢组织完整性。结果 研究发现,2%浓度海藻酸钠水凝胶包封卵巢组织冷冻后的卵泡存活率最高。先加载保护剂再进行包封的方式更适合卵巢组织冷冻保存,并能将所需冷冻保护剂浓度降低至原浓度的50%。8 g/L Fe3O4磁纳米粒子与300 Gs的交变磁场对卵巢组织的复温效果最佳。水浴结合磁热复温卵巢组织后卵泡存活率最高,抗氧化能力最好且组织形态得到了良好的保持。结论 海藻酸钠水凝胶包封卵巢组织能够在冷冻环节中降低保护剂浓度,磁热与水浴的结合使用能够更高效地实现卵巢组织的复温。本研究为卵巢组织玻璃化冷冻及复温的优化提供了新的方法。
Objective For prepubertal and urgently treated malignant tumor patients, ovarian tissue cryopreservation and transplantation represent more appropriate fertility preservation methods. Current clinical practices often involve freezing ovarian tissue with high concentrations of cryoprotectants (CPAs) and thawing with water baths. These processes lead to varying degrees of toxicity and devitrification damage to ovarian tissue. Therefore, this paper proposes optimized methods for vitrification of ovarian tissues based on sodium alginate hydrogel encapsulation and magnetic induction nanowarming technology.Methods Firstly, the study investigated the effects of sodium alginate concentration, the sequence of hydrogel encapsulation and CPAs loading on vitrification efficiency of encapsulated ovarian tissue. Additionally, the capability of sodium alginate hydrogel encapsulation to reduce the required concentration of CPAs was validated. Secondly, a platform combining water bath and magnetic induction nanowarming was established to rewarm ovarian tissue under various concentrations of magnetic nanoparticles and magnetic field strengths. The post-warming follicle survival rate, antioxidant capacity, and ovarian tissue integrity were evaluated to assess the efficacy of the method.Results The study found that ovarian tissue encapsulated with 2% sodium alginate hydrogel exhibited the highest follicle survival rate after vitrification. The method of loading CPAs prior to encapsulation proved more suitable for ovarian tissue cryopreservation, effectively reducing the required concentration of CPAs by 50%. A combination of 8 g/L Fe3O4 nanoparticles and an alternating magnetic field of 300 Gs showed optimal warming effectiveness for ovarian tissue. Combining water bath rewarming with magnetic induction nanowarming yielded the highest follicle survival rate, enhanced antioxidant capacity, and preserved tissue morphology.Conclusion Sodium alginate hydrogel encapsulation of ovarian tissue reduces the concentration of CPAs required during the freezing process. The combination of magnetic induction nanowarming with water bath provides an efficient method ovarian tissue rewarming. This study offers novel approaches to optimize ovarian tissues vitrification.
曹裕坤,叶娜,李铮,周新丽.水凝胶包封及磁热复温优化卵巢组织玻璃化保存[J].生物化学与生物物理进展,2025,52(2):464-477
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