PEEK-SPEEK-HA复合材料与U2OS细胞相容性研究

深圳大学化学与环境工程学院,广东深圳518060

聚醚醚酮-羟基磷灰石复合材料; U2OS骨瘤细胞; 细胞增殖; 细胞形态; 细胞黏附; 细胞凋亡; 活性氧

Experiments on biocompatibility of PEEK-SEEK-HA composite materials with U2OS cells
Ni Zhuo,Huang Weiyin,Wang Ying,Wu Gengfeng,and Pan Tianguang

College of Chemistry and Environmental Engineering, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China

polyetheretherketone-hydroxyapatite(PEEK-HA)composite; U2OS cell; cell morphology; proliferation; cell adhesion; cell apoptosis; reactive oxygen species(ROS)

DOI: 10.3724/SP.J.1249.2016.01001

备注

体外培养U2OS细胞,通过噻唑蓝(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet razolium bromide, MTT)法研究新型聚醚醚酮-羟基磷灰石(polyetheretherketone-hydroxyapatite, PEEK-HA)生物复合材料对细胞增殖的影响.研究发现,聚醚醚酮-磺化聚醚醚酮-羟基磷灰石(polyetheretherketone-sulfnated polyetheretherketone-hydroxyapatite, PEEK-SPEEK-HA)复合材料的细胞增殖作用较PEEK-HA复合材料强,且随着HA含量的增加细胞增殖作用增强,材料浸提液质量浓度为4~8 mg/mL时对细胞有较好的增殖促进作用.复合材料毒性级别为Ⅰ,对细胞无毒性.测定黏附率研究该材料对U2OS细胞的黏附特性,PEEK-SPEEK-HA复合材料随着HA含量的增大,细胞黏附率增加,高于PEEK-HA复合材料组,表明加入HA和SPEEK能改善复合材料的黏附性能.采用流式细胞术研究材料对细胞凋亡的影响,发现加入SPEEK的PEEK-HA复合材料能抑制细胞发生早期凋亡,从而降低凋亡率.激光共聚焦和流式细胞仪检测该材料对细胞活性氧的影响发现,SPEEK引入复合材料能够降低细胞活性氧水平,降低细胞毒性. 扫描电镜实验表明,U2OS细胞能够在PEEK-SPEEK-HA复合材料表面黏附、伸展和生长.

Effects of polyetheretherketone-hydroxyapatite(PEEK-HA)composites and polyetheretherketone-sulfnated polyetheretherketone-hydroxyapatite(PEEK-SPEEK-HA)composites on U2OS cells are studied by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet razolium bromide(MTT)method. It is found that cell proliferation in PEEK-SPEEK-HA composites is broader than that in PEEK-HA composites. With an increase of HA content in PEEK-HA composites, cell proliferation accelerates. The material of sulfnated polyetheretherketone promotes cell proliferation significantly at 4-8 mg/mL concentration. A cytotoxicity test shows that these bio-materials are non-toxic to U2OS cells and the U2OS cells can adapt to these materials. With an increase of HA content, the cell adhesion rate increases and is higher in PEEK-SPEEK-HA composites than in PEEK-HA composites, which indicates that adding HA and SPEEK in PEEK composite could improve the adhesion property. Cell apoptosis test by the flow cytometry(FCM)indicates that the addition of SPEEK in PEEK-HA composites can inhibit the early apoptosis thus decreasing the apoptosis rate. A test on the effect of SPEEK on the reactive oxygen species(ROS)of U2OS cells by using the flow cytometry(FCM)and the confocal laser scanning microscopy(CLSM)shows that SPEEK in PEEK-HA composites can decrease the ROS level, and reduce the cytotoxicity. Scanning electron microscope(SEM)results show that U2OS cells can adhere, stretch and grow on surfaces of PEEK-SPEEK-HA composites.

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