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驻极体静电场对巨噬细胞迁移能力的影响
梁媛媛1,涂晔2,崔黎丽3,江键1*
0
(1. 海军军医大学(第二军医大学)卫生勤务学系数理教研室, 上海 200433;
2. 上海市东方医院学科办, 上海 200120;
3. 海军军医大学(第二军医大学)药学系无机化学教研室, 上海 200433
*通信作者)
摘要:
目的 探讨驻极体产生的静电场对巨噬细胞迁移能力的影响。方法 常温下利用低温等离子体放电法(栅控恒压电晕放电)制备得-2 000 V聚丙烯驻极体,借助常规等温表面电位衰减测量方法研究驻极体在储存和实验条件下的电荷储存稳定性。用紫外线消毒后的驻极体作用于对数生长期的小鼠巨噬细胞(RAW264.7细胞),采用细胞划痕实验、Transwell细胞实验、荧光探针示踪法研究在驻极体静电场作用下细胞的迁移和形态变化。结果 -2 000 V驻极体在常温、常湿条件下放置14 d时其表面电位稳定在初始值的70%,置于细胞培养箱28 h后其表面电位与0 h时相比差异无统计学意义(P>0.05)。细胞划痕实验结果显示,驻极体静电场作用28 h时RAW264.7细胞向划痕区迁移、增殖能力增强,划痕面积减小。Transwell实验结果显示,驻极体静电场作用12和24 h时跨膜细胞数均较对照组增加(P均<0.01)。荧光探针示踪实验结果显示,驻极体静电场作用下RAW264.7细胞体积变大,发生延展,褶皱增多,生出板状伪足、丝状伪足等突足结构。结论 制备的-2 000 V驻极体可在体外提供稳定的静电场并持续作用于细胞,其静电场促进巨噬细胞体积变大、发生延展、褶皱增多并生出板状伪足和丝状伪足等突足结构,增强了巨噬细胞的迁移能力。
关键词:  静电场  驻极体  巨噬细胞  细胞迁移  伤口愈合
DOI:10.16781/j.0258-879x.2021.06.0688
投稿时间:2020-12-25修订日期:2021-04-09
基金项目:上海市自然科学基金(19ZR1469300),上海市卫生和计划生育委员会青年科学基金(20164Y0051).
Effect of electret electrostatic field on migration of macrophages
LIANG Yuan-yuan1,TU Ye2,CUI Li-li3,JIANG Jian1*
(1. Department of Physics and Mathematics, Faculty of Health Service, Naval Medical University (Second Military Medical University), Shanghai 200433, China;
2. Department of Medical Affairs, Shanghai East Hospital, Tongji University, Shanghai 200120, China;
3. Department of Inorganic Chemistry, School of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, China
*Corresponding author)
Abstract:
Objective To investigate the effect of electret electrostatic field on migration of macrophages. Methods The -2 000 V polypropylene electret was prepared by low temperature plasma discharge (grid controlled constant voltage corona discharge) at room temperature. The charge storage stability of the electret under storage and experimental conditions was studied by conventional isothermal surface potential decay measurement. The mouse macrophages (RAW264.7 cells) in logarithmic growth phase were treated by ultraviolet sterilized electret. The cell migration and morphological changes under the electret electrostatic field were analyzed by cell scratch test, Transwell assay and fluorescent probe tracing. Results The surface potential of -2 000 V electret was stable at 70% of its initial value when it was placed at room temperature and normal humidity for 14 d. There was no significant difference between the surface potential of -2 000 V electret in cell incubator for 28 h and that for 0 h (P>0.05). The results of the cell scratch test showed that after exposed to the electret electrostatic field for 28 h, the migration and proliferation ability of RAW264.7 cells to scratch area was enhanced, and the scratch area decreased. The results of the Transwell assay showed that the number of transmembrane cells in electret treated group increased compared with the control group at 12 and 24 h (both P<0.01). The results of the fluorescent probe tracing showed that RAW264.7 cells became larger, expanded, with increased ruffling, and produced protuberant structures such as plate-shaped pseudopodia and filamentous pseudopodia under the action of negative electret. Conclusion The prepared -2 000 V electret can provide a stable electrostatic field in vitro and continuously act on cells. The electrostatic field can promote the macrophages to become larger, extended, with increased ruffling, and produce protuberant structures such as plate-shaped pseudopodia and filamentous pseudopodia, which can improve the migration ability of macrophages.
Key words:  electrostatic field  electret  macrophages  cell migration  wound healing