自组装金片表面化学功能团对骨肉瘤细胞黏附的影响

Effects of chemical groups of gold surface with self-assembled monolayers on osteosarcoma cells adhesion

目的探讨自组装金片表面化学功能团对骨肉瘤细胞黏附的影响。方法将标准统一的金片,放入带有1%不同功能团的硫醇试剂中浸泡12 h,自组装为甲基、氨基、羧基和羟基功能团的单一表面膜,并对其表面接触角进行测定。将U2-OS细胞接种于不同功能团及对照组(裸金片)表面,培养3、6、9 h后,光镜下观察各组细胞黏附数及形态学变化,选择培养6 h的金片进行扫描电镜观察。结果羟基、羧基、氨基和甲基表面接触角为9.5°±1.2°、18.3°±3.9°、59.7°±3.8°和105°±9.1°,不同化学基团修饰表面的接触角比较,差异有统计学意义(P<0.05)。培养3 h,各组黏附于金片表面的细胞基本呈小圆形,甲基表面黏附的细胞不紧密,呈将脱落状;培养6 h,氨基、羧基、羟基及对照组金片表面细胞体积增大,可见大椭圆形、梭形及多边形细胞,而在甲基组金片表面细胞仍呈小圆形;培养9 h,各组细胞形态变化差异更加明显,而甲基表面的细胞多数仍呈圆形或椭圆形。随着培养时间的延长,甲基金片表面的细胞数增加最不明显,不同时相点各组细胞数比较,差异有统计学意义(P<0.05);各组金片表面黏附细胞数总体趋势为甲基<<羟基<羧基≈氨基。扫描电镜下细胞在羟基和羧基表面多为椭圆形和多边形,细胞体积较大,胞浆丰富,可见较多伪足伸出,部分较大伪足相互融合;氨基表面细胞多数为长梭形;甲基表面的细胞仍呈小圆形,细胞体积较小。结论甲基功能团可能对自组装金片表面骨肉瘤细胞的黏附行为具有抑制作用。

Objective To observe the effects of chemical groups of gold surface with self-assembled monolayers （SAMs） on U2-OS sarcoma cells adhesion. Methods After immersion into thiol solutions with 1%different chemical groups for 12 h respectively, the methyl （-CH3）, amino （-NH2）, carboxyl （-COOH） and hydroxyl （-OH） functional groups were self-assembled on Au layers, and the surface contact angles were determined respectively to evaluate the equivalent surface densities of chemical functional groups. U2-OS cells were seeded on these substrates, and the surface of bare Au layers was as control. Cell adhesion was observed under light microscope after 3, 6, 9 hours＆#39; culture, and further examination was undertaken by scanning electron microscopy （SEM） after 6 hours＆#39; culture. Results The contact angle of-OH,-COOH,-NH2 and-CH3 surface was 9.5° ± 1.2° , 18.3° ± 3.9° , 59.7° ± 3.8° and 105° ± 9.1° respectively, which there had statistical differences among these chemical groups （P 〈0.05）. Under light microscope, after 3 hours＆#39; culture, almost all of U2-OS cells adhered onto the surface of the gold subtrates began to present a small round shape, the cells on the-CH3 surface seemed to be very loose. Six hours later, cells volume on the surface of-NH2,-COOH,-OH and control group increased, and large oval, polygonal, spindle cells had been found, but the cells on-CH3 surface were still small round. After 9 hours＆#39; incubation, there existed significant differences of cell morphology in different groups, while cells on-CH3 surface still remained round or oval. As the culture time went by, cell number increase on-CH3 surface was the least, the differences of cell number among groups at each time point had statistical significance （P 〈0.05）; The adhesion number of U2-OS cells followed the trend：-CH3〈〈-OH〈-COOH≈-NH2. SEM results indicated that U2-OS cells cultured on-OH and-COOH surfaces after 6 hours＆#39; incubation often exhibited polygonal and oval morphology with bigger volume and adbundant cytoplasm, and more pseudopodia extended and partial fused with each other; Most of cells on-NH2 surface showed a spindle shape, while cells cultured on-CH3 surface were smaller and in a spherical shape. Conclusion-CH3 functional group probably inhibit the adhesion of osteosarcoma cells on the gold surface with SAMs.