CAJ | 학술논문

为了将聚二甲基硅氧烷(PDMS)用作空间微流控细胞培养芯片的结构材料,需要克服其自身不利于细胞黏附以及容易吸附蛋白质的缺陷,故必须通过简单可靠的表面处理方法改变PDMS的表面特性.为此,以航天医学生物学研究中常用人骨肉瘤细胞MG-63的黏附和细胞培养液中常见蛋白质牛血清白蛋白(BSA)的吸附为衡量指标,比较了 4 种 PDMS 表面处理方法的效果,并初步研究了 PDMS 表面浸润性及形貌与 MG-63 黏附/BSA 吸附之间的关系.4 种方法分别为在 PDMS 表面自聚合聚多巴胺(PDA-PDMS)、涂覆聚赖氨酸(PLL-PDMS)、包被胶原蛋白Ⅰ型(COL-PDMS)和氧等离子体处理(ox-PDMS).用倒置荧光显微镜观察细胞黏附和铺展,并用CellTiter 96? AQueous单溶液细胞增殖检测试剂盒(MTS)检测培养 1~4,d 的 MG-63 活细胞活性;检测了吸附异硫氰酸荧光素标记牛血清白蛋白(FTIC-BSA)后的 PDMS 表面荧光强度;测量了处理前后 PDMS 表面接触角及表面形貌.结果表明:PDA-PDMS和ox-PDMS变得亲水且较粗糙,而PLL-PDMS、COL-PDMS和未处理PDMS疏水且较平滑;MG-63经4种方法处理后,PDMS 表面均能较好地黏附和聚二甲基硅氧烷铺展,但在亲水表面黏附更持久;BSA 在亲水表面的吸附明显少于憎水表面.
위료장취이갑기규양완(PDMS)용작공간미류공세포배양심편적결구재료,수요극복기자신불리우세포점부이급용역흡부단백질적결함,고필수통과간단가고적표면처리방법개변PDMS적표면특성.위차,이항천의학생물학연구중상용인골육류세포MG-63적점부화세포배양액중상견단백질우혈청백단백(BSA)적흡부위형량지표,비교료 4 충 PDMS 표면처리방법적효과,병초보연구료 PDMS 표면침윤성급형모여 MG-63 점부/BSA 흡부지간적관계.4 충방법분별위재 PDMS 표면자취합취다파알(PDA-PDMS)、도복취뢰안산(PLL-PDMS)、포피효원단백Ⅰ형(COL-PDMS)화양등리자체처리(ox-PDMS).용도치형광현미경관찰세포점부화포전,병용CellTiter 96? AQueous단용액세포증식검측시제합(MTS)검측배양 1~4,d 적 MG-63 활세포활성;검측료흡부이류청산형광소표기우혈청백단백(FTIC-BSA)후적 PDMS 표면형광강도;측량료처리전후 PDMS 표면접촉각급표면형모.결과표명:PDA-PDMS화ox-PDMS변득친수차교조조,이PLL-PDMS、COL-PDMS화미처리PDMS소수차교평활;MG-63경4충방법처리후,PDMS 표면균능교호지점부화취이갑기규양완포전,단재친수표면점부경지구;BSA 재친수표면적흡부명현소우증수표면.
As the native poly(dimethylsiloxane)(PDMS)is unfavorable for cell adhesion and easy to adsorb proteins,the PDMS should be modified for cell-culture applications. In this study,the effectiveness of four simple PDMS surface modification methods for cell culture has been compared with the adhesion of MG-63 cells and the amount of bovine serum albumin(BSA)adsorption as indicators. The relationship between PDMS' surface wettability and topography and the MG-63 adhesion/BSA adsorption was studied. Four groups of modified substrates included PDMS coated with poly(dopamine)(PDA-PDMS),poly-L-lysine(PLL-PDMS),collagen typeⅠ(COL-PDMS), respectively,and oxidized using oxygen plasma(ox-PDMS). The MG-63 adhesion and spreading were observed using an inverted fluorescence microscope,and the attached MG-63 at 1—4,dwere quantitatively evaluated by CellTiter 96? AQueous one solution cell proliferation assay(MTS). The intensities of fluorescein isothiocyanate-bovine serum albumin(FITC-BSA)adsorbed on all groups of PDMS substrates were detected. The native/treated PDMS' surface contact angle and topography were also measured. The experimental results reveal that PDA-PDMS and ox-PDMS surfaces become hydrophilic and rough,while PLL-PDMS,COL-PDMS and native PDMS surfaces are still hydrophobic and smooth. MG-63 prefers to attach and spread on all the four treated PDMS substrates,andstays longer on the hydrophilic surfaces. BSA adsorption on the hydrophilic surfaces is significantly less than that on the hydrophobic surfaces.