静态压应力促进三维培养下的人牙周膜细胞自噬  

Static compressive stress accelerting autophagy of human ligament cells in three-dimensional culture

作　　者：吴诗阳 亢雪萍 邹蕊[1] 柴桢 吉玲玲[1] WU Shiyang;KANG Xueping;ZOU Rui;CHAI Zhen;JI Lingling(Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research,College of Stomatology,Laboratory Center of Stomatology,College of Stomatology,Xi'an Jiaotong University,China,710004)

机构地区：[1]西安交通大学口腔医院,陕西省颅颌面精准医学研究重点实验室,陕西省牙颌疾病临床研究中心,710004

出　　处：《实用口腔医学杂志》2025年第2期189-192,共4页Journal of Practical Stomatology

基　　金：陕西省自然科学基础研究计划(编号:2023-JC-YB-813)。

摘　　要：目的:建立一个体外三维培养和静态压应力加载的人牙周膜细胞(hPDLCs)模型,并检测静态压应力对细胞自噬的影响。方法:从新鲜的离体牙表面刮取牙周组织进行培养,获得hPDLCs;提取Ⅰ型鼠尾胶原,制备胶原-藻酸盐复合水凝胶,将hPDLCs包埋进水凝胶中,培养72 h后加力,使用实时定量聚合酶链式反应(qRT-PCR)检测自噬相关蛋白LC3,Atg-5,Beclin-1的基因表达,使用蛋白印迹(Western blot)检测Atg-5,Beclin-1及LC3的蛋白表达。结果:细胞在胶原-藻酸盐水凝胶中生长良好;在压应力加载15 min后,hPDLCs自噬水平显著提高,之后逐渐降低到基线水平。结论:胶原方木藻酸盐水凝胶具有良好的生物相容性与安全性,可作为细胞体外三维培养的优良载体;静态压应力可以促进细胞自噬,且这种促进作用是迅速而短暂的。Objective:To establish a three-dimensional culture and stress-loaded model of human periodontal ligament cells(hPDLCs)in vitro and to determine the effect of static pressure on autophagy.Methods:Periodontal tissue was scraped from fresh extracted tooth surfaces for culture and hPDLCs were obtained.Type I rat tail collagen was extracted to prepare collagen-sodium alginate composite hydrogel,in which third-generation hPDLCs were embedded.After 72 h of culture,gene expression of autophagy-related proteins LC3,Atg-5 and Beclin-1 was detected by quantitative real-time polymerase chain reaction(qRT-PCR),and the e xpression of Atg-5,Beclin-1 and LC3 was detected by Western blotting.Results:Cells grew well in collagen-alginate hydrogels.After 15 min of compressive stress loading,the level of autophagy of hPDLCs increased significantly and then gradually decreased to the baseline level.Conclusion:Collagen-alginate hydrogel has good biocompatibility and safety,and can be used as an excellent scaffold for three-dimensional cell culture in vitro.Stati c compressive stress can accelevate autophagy,and this acceleration is rapid and short-lived.

关 键 词：细胞自噬 胶原-藻酸盐复合水凝胶 人牙周膜细胞 静态压应力 

分 类 号：R781.4[医药卫生—口腔医学]