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目的观察三维回转模拟失重对人牙周膜成纤维细胞(human periodontal ligament fibroblasts,hP-DLFs)形态和功能的影响。方法回转组hPDLF细胞运用三维回转器(TDC)分别培养48 h,72 h和96 h,对照组常规培养。利用HE染色、细胞计数法、MTT活性检测、流式细胞术、Hoechst 33258核染色及免疫荧光分析的方法,比较两组细胞间形态、增殖、周期、凋亡及细胞骨架等生物学性能的差异。结果与各自对照组相比,hPDLF细胞三维回转培养48 h后,细胞核面积较对照组缩小(P<0.01),微丝结构略显模糊,但细胞增殖和凋亡无明显变化;回转72 h后,细胞活力显著增强(P<0.01),但细胞形态与凋亡无明显变化;回转96 h后,hPDLF细胞增殖能力较对照组显著增强(P<0.01),细胞活力明显升高(P<0.01),S期所占周期比例明显增加(P<0.05),细胞形态与凋亡与对照组比较仍无明显差异。结论短期(48 h)暴露于三维回转培养条件使hPDLF细胞骨架结构和胞核形态发生变化,但随着刺激时间的延长(72 h后),细胞内部恢复正常,并表现出细胞活力及增殖能力增强的趋势。
Objective To observe the effects of three-dimensional swirling simulated weightlessness on the morphology and function of human periodontal ligament fibroblasts (hP-DLFs). Methods The hPDLF cells in the swivel group were cultured for 48 h, 72 h and 96 h with three-dimensional rotator (TDC), respectively. The control group was cultured routinely. The morphological, proliferation, cell cycle, apoptosis and cytoskeleton of the two groups were compared by HE staining, cell counting method, MTT assay, flow cytometry, Hoechst 33258 nuclear staining and immunofluorescence analysis . Results Compared with the control group, the hPDLF cells in three-dimensional rotary culture for 48 h had a smaller nucleus area than the control group (P <0.01), and the fibroblast structure was slightly vague, but the cell proliferation and apoptosis did not change significantly. After 72 h of revolving, (P <0.01), but there was no significant change in cell morphology and apoptosis. After 96 h of rotation, the proliferation of hPDLF cells was significantly enhanced compared with the control group (P <0.01), and the cell viability was significantly increased (P <0.01) ), The proportion of S phase was significantly increased (P <0.05), cell morphology and apoptosis compared with the control group still no significant difference. Conclusion Short-term (48 h) exposure to three-dimensional rotary culture conditions changed the cytoskeleton and nuclear morphology of hPDLF cells. However, with prolonged stimulation (after 72 h), the cells returned to normal and exhibited cell viability and proliferation Increased trend.