摘要
采用细胞与材料直接作用的方式研究钛酸锶微/纳米颗粒刺激RAW264.7细胞产生的生物学反应。利用扫描电子显微镜(scanning electron microscope, SEM)、激光粒度仪、X射线衍射仪及比表面积测定仪表征材料的理化性质。采用CCK-8检测和活/死细胞染色法评估钛酸锶微/纳米颗粒对RAW264.7细胞增殖活性的影响;采用SEM观察细胞的形态变化;采用RT-qPCR技术检测在钛酸锶微/纳米颗粒刺激作用下RAW 264.7细胞炎性细胞因子基因的表达;采用免疫荧光染色技术检测RAW264.7细胞诱导型一氧化氮合酶(induciblenitricoxidesynthase,iNOS)的表达。结果表明,微米和纳米钛酸锶颗粒成分组成均一,皆为不规则形状,微米钛酸锶颗粒平均粒径为1 085.0 nm,比表面积为1.72 m^(2)/g,纳米钛酸锶颗粒平均粒径为505.2 nm,比表面积为2.94 m^(2)/g。微米和纳米钛酸锶颗粒对RAW 264.7细胞第24 h、72 h活性并无影响。在微/纳米钛酸锶颗粒的刺激下RAW 264.7细胞向多边形发展,并具有更大的贴附面积。微米钛酸锶颗粒刺激RAW 264.7细胞高表达iNOS、白细胞分化抗原(cluster of differentiation, CD)86、肿瘤坏死因子(tumor necrosis factor, TNF)-α和白细胞介素(Interleukin,IL)-6基因。纳米钛酸锶颗粒刺激RAW264.7细胞高表达iNOS和TNF-α基因,而IL-10基因的表达显著下调。微米和纳米均刺激RAW264.7细胞高表达iNOS,诱导RAW264.7细胞向M1表型极化,产生炎症反应,并且微米钛酸锶颗粒诱导的炎症反应更显著。
The biological response of RAW 264.7 cells stimulated by strontium titanate micro/nanoparticles was studied by the direct interaction between cells and materials. Scanning electron microscope(SEM), laser particle size analyzer,X-ray diffractometer and specific surface area tester were used to characterize the physical and chemical properties of the materials. The effects of strontium titanate micro/nanoparticles on the proliferation activity of RAW 264.7 cells were evaluated by CCK-8 test and live/dead cell staining;the morphological changes of cells were observed by SEM;The expression of inflammatory cytokine genes in RAW 264.7 cells stimulated by strontium acid micro/nanoparticles. The results show that the compositions of the micron and nano strontium titanate particles are uniform and both have irregular shapes. The average particle size of the micron strontium titanate particles is 1 085.0 nm, the specific surface area is 1.72 m^(2)/g, and the particle size of the nano strontium titanate particles is 505.2 nm. The specific surface area is 2.94 m^(2)/g. The micron and nano strontium titanate particles have no effect on the proliferation activity of RAW 264.7 cells on the 24 h and 72 h. Under the stimulation of micro/nano strontium titanate particles, the RAW 264.7 cell morphology develops into a polygonal shape and has a larger attachment area. Micron strontium titanate particles stimulated high expression of inducible nitric oxide synthase(iNOS), cluster of differentiation(CD) 86, TNF-α and interleukin(IL)-6 genes in RAW 264.7 cells, and the expression of iNOS, tumor necrosis factor(TNF)-α and IL-6 genes is significantly higher than that of nano strontium titanate particles. nano strontium titanate particles stimulated RAW 264.7 cells to highly express iNOS and TNF-α genes, while IL-10 gene expression is significantly down-regulated. Both micron and nano strontium titanate particles stimulate RAW 264.7 cells to highly express iNOS, induce RAW 264.7 cells to polarize to the M1 phenotype,and produce an inflammatory response. The inflammatory response induced by micron strontium titanate particles is more significant.
作者
刘文涛
吴宏
周吉祥
LIU Wentao;WU Hong;ZHOU Jixiang(State Key Laboratory of Powder Metallurgy,Central South University,Changsha 410083,China;Xiangya Hospital,Central South University,Changsha 410083,China)
出处
《粉末冶金材料科学与工程》
2021年第5期475-482,共8页
Materials Science and Engineering of Powder Metallurgy
基金
国家自然科学基金资助项目(52071346)
长沙市自然科学基金资助项目(kq2014293)。
