中国组织工程研究2026，Vol.30Issue(30)：7792-7800,9.DOI:10.12307/2026.963
EVs@ZIF-8载药体系在成骨细胞自噬抑制环境下促进MC3T3-E1早期成骨分化
The EVs@ZIF-8 drug-loaded system promotes early osteogenic differentiation of MC3T3-E1 cells under autophagy-inhibited conditions in osteoblasts
摘要
Abstract
BACKGROUND:Autophagy is an essential step in the mineralization of osteoblasts in vitro,and dysregulation of autophagy activity can affect the mineralization ability of osteoblasts.Exosomes can promote osteogenesis by regulating autophagy to eliminate harmful substances.However,how to stably transport and release exosomes remains challenging. OBJECTIVE:To investigate whether the successfully synthesized extracellular vesicles(EVs)@zeolitic imidazolate framework-8(ZIF-8)drug-loaded system has an osteogenic effect and whether autophagy may be involved in this process. METHODS:(1)Material synthesis:The plasma exosomes were extracted by an extraction and purification kit,and the successful extraction of exosomes was verified by western blot.ZIF-8 was synthesized via a hydration method,followed by the one-pot synthesis of the EVs@ZIF-8 drug-loaded system.The nano-drug-loaded system was characterized and tested using scanning electron microscopy and X-ray diffraction.(2)Materials promote osteogenesis:MC3T3-E1 cells were co-cultured with different concentrations of EVs@ZIF-8 and exosome groups,and cell proliferation and viability assays were conducted to verify the activity of exosomes and screen for the optimal concentration of EVs@ZIF-8.Subsequently,cells were co-cultured with blank,ZIF-8,and EVs@ZIF-8 groups.After osteogenic induction,alkaline phosphatase detection was used to verify the early osteogenic differentiation ability of MC3T3-E1 promoted by the drug-loaded system.The expression levels of osteoprotegerin,Runx2,and type Ⅰ collagen genes in the samples were measured.(3)Materials promote osteogenesis via autophagy:MC3T3-E1 cells were co-cultured with different concentrations of autophagy inhibitor 3-methyladenine,and the optimal concentration of 3-methyladenine was determined through cell proliferation and viability assays.Subsequently,cells were co-cultured with 3-methyladenine,3-methyladenine+ZIF-8,and 3-methyladenine+EVs@ZIF-8 groups.After osteogenic induction,alkaline phosphatase detection was performed to verify the early osteogenic differentiation ability of MC3T3-E1 promoted by the drug-loaded system. RESULTS AND DISCUSSION:(1)Under scanning electron microscopy,EVs@ZIF-8 exhibited a uniform spherical encapsulation structure with a particle size of(232.5±22.0)nm.(2)The results of cell proliferation and viability experiments indicated that 50 μg/mL EVs@ZIF-8 significantly promoted the proliferation of MC3T3-E1 cells,while 5.0 mmol/L 3-methyladenine exerted its function without excessively inhibiting cell proliferation.Therefore,the aforementioned concentrations were selected for subsequent cell experiments.(3)After osteogenic induction,the alkaline phosphatase activity and osteogenic gene expression in the EVs@ZIF-8 group were higher than those in the other two groups(P＜0.05).After the addition of 3-methyladenine,an autophagy inhibitor,the proliferation,alkaline phosphatase activity,and osteogenic gene expression in the EVs@ZIF-8 group were all still higher than those in the other two groups.These results indicated that cell proliferation was inhibited in the presence of autophagy inhibitors and EVs@ZIF-8 partially antagonized this inhibitory effect,suggesting that autophagy may be involved in the process by which the EVs@ZIF-8 drug delivery system promotes osteogenic differentiation of cells.关键词
EVs@ZIF-8/自噬/成骨分化/细胞外囊泡/成骨细胞Key words
EVs@ZIF-8/autophagy/osteogenic differentiation/extracellular vesicles/osteoblasts分类
医药卫生引用本文复制引用
郭珈伶,薛轶元,裴锡波..EVs@ZIF-8载药体系在成骨细胞自噬抑制环境下促进MC3T3-E1早期成骨分化[J].中国组织工程研究,2026,30(30):7792-7800,9.基金项目
国家自然科学基金(82471021),课题名称:基于锌-铁离子平衡策略构建巨噬细胞靶向递送系统治疗糖尿病种植体周围炎的机制研究,项目负责人:裴锡波 （82471021）
国家自然科学基金-青年基金项目(82301150),课题名称:自噬介导年轻BMSCs-EVs@ZIF-8促进老年性骨质疏松颌骨缺损修复的机制研究,项目负责人:薛轶元 National Natural Science Foundation of China,No.82471021(to PXB) （82301150）
National Natural Science Foundation of China-Youth Fund Project,No.82301150(to XYY) （to XYY）
