CAJ | 학술논문

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6h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。
목적：탐토자조장금편표면화학공능단대골육류세포점부적영향。방법장표준통일적금편，방입대유1%불동공능단적류순시제중침포12 h，자조장위갑기、안기、최기화간기공능단적단일표면막，병대기표면접촉각진행측정。장U2-OS세포접충우불동공능단급대조조（라금편）표면，배양3、6、9 h후，광경하관찰각조세포점부수급형태학변화，선택배양6h적금편진행소묘전경관찰。결과간기、최기、안기화갑기표면접촉각위9.5°±1.2°、18.3°±3.9°、59.7°±3.8°화105°±9.1°，불동화학기단수식표면적접촉각비교，차이유통계학의의（P＜0.05）。배양3 h，각조점부우금편표면적세포기본정소원형，갑기표면점부적세포불긴밀，정장탈락상；배양6 h，안기、최기、간기급대조조금편표면세포체적증대，가견대타원형、사형급다변형세포，이재갑기조금편표면세포잉정소원형；배양9 h，각조세포형태변화차이경가명현，이갑기표면적세포다수잉정원형혹타원형。수착배양시간적연장，갑기금편표면적세포수증가최불명현，불동시상점각조세포수비교，차이유통계학의의（P＜0.05）；각조금편표면점부세포수총체추세위갑기<<간기<최기≈안기。소묘전경하세포재간기화최기표면다위타원형화다변형，세포체적교대，포장봉부，가견교다위족신출，부분교대위족상호융합；안기표면세포다수위장사형；갑기표면적세포잉정소원형，세포체적교소。결론갑기공능단가능대자조장금편표면골육류세포적점부행위구유억제작용。
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' culture, and further examination was undertaken by scanning electron microscopy (SEM) after 6 hours' 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' 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' 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' 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.