微阵列技术探究限域面积对成牙本质细胞极化和分化的影响
Effects of cell area on single odontoblast polarization and differentiation via microarray technology
作者：李虎恩, 于年祚, 李熙恒, 唐晓铎, 孙雅鹿, 司超, 张俊虎, 常蓓
Author：Li Huen, Yu Nianzuo, Li Xiheng, Tang Xiaoduo, Sun Yalu, Si Chao, Zhang Junhu, Chang Bei
收稿日期：2024-10-24 年卷（期）页码：2025,43(2):183-183-189
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：细胞极化,图案化,成牙本质细胞,细胞分化,
Key words：cell polarization,micropatterning,odontoblast,cell differentiation,
基金项目：国家自然科学基金青年基金(82101075);中国科学技术协会青年人才托举项目(YESS20210407);博士后科学基金面上项目(2022M711301);吉林大学“白求恩计划”(419161923025)
中文摘要
目的 利用光刻技术制备图案化的细胞差异黏附表面，探讨黏附面积对成牙本质细胞极化与成牙向分化的影响。 方法 利用光刻技术和聚乙二醇二丙烯酸酯（PEGDA）水凝胶制备了具有差异黏附特性的图案化微阵列，通过对细胞产生限域作用，促使人牙髓干细胞（hDPSCs）呈现天然成牙本质细胞样形态。筛选孤立培养的单个hDPSCs，探究不同面积（1 800、2 700、3 600 μm2）对成牙本质细胞极化和分化的影响。通过免疫荧光染色观察细胞形态，明确高尔基体和细胞核在hDPSCs中的定位及取向，分析不同面积对细胞极化的作用。通过碱性磷酸酶染色检测hDPSCs成牙向分化率和平均光密度值，探究不同面积对细胞分化的作用。 结果 成功分离了hDPSCs并对其鉴定，制备了仿成牙本质细胞形态的图案化微阵列，通过鬼笔环肽染色证实了细胞形状与微图案形状高度相符。免疫荧光染色结果显示限域面积为3 600 μm2时，孤立hDPSCs中各项极化和分化水平均明显高于1 800 μm2和2 700 μm2限域面积的孤立hDPSCs（P<0.05）。 结论 本实验通过微阵列技术证实了限域面积参与调控成牙本质细胞极化和分化进程，随着限域面积增大，孤立hDPSCs极化和分化水平升高。
英文摘要
ObjectiveThis study aimed to explore the impact of cell spreading area on odontoblast polarization and differentiation using micropatterned surfaces ge-nerated by photolithography.MethodsMicropatterned surfaces with differential adhesive properties were prepared using polyethylene glycol diacrylate (PEGDA)-ba-sed photolithography. Human dental pulp stem cells (hD-PSCs) were isolated into single cells and cultured on micropatterned surfaces with areas of 1 800, 2 700, and 3 600 μm2. Immunofluorescence staining was used to observe cell morphology and analyze the relocating of the golgi apparatus and nucleus. Alkaline phosphatase staining was preformed to examine odontogenic differentiation.ResultsThe hDPSCs were successfully isolated and cultured on micropatterned surfaces mimicking the morphology of polarized odontoblasts. Phalloidin staining confirmed that the isolated hDPSCs successfully recapitulated the morphology of predesigned micropatterns. Immunofluorescence staining showed that the polarization and differentiation levels of the hDPSCs with a 3600 μm2area were significantly higher than those with 1 800 and 2 700 μm2areas (P<0 .05).ConclusionThe polarization and differentiation of single hDPSCs increased with the cell areas on micropatterned surfaces.
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