基底刚度对成纤维细胞生物学行为的影响  

Effect of substrate stiffness on biological behavior of fibroblasts

作　　者：王玉[1] 王贵学[1] 罗向东[2] 邱菊辉[1] 

机构地区：[1]重庆大学生物工程学院,生物流变科学与技术教育部重点实验室,400030 [2]第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室

出　　处：《中华烧伤杂志》2011年第6期427-431,共5页Chinese Journal of Burns

基　　金：国家自然科学基金（30730093、30970721）

摘　　要：目的 了解基底刚度对FH增殖、迁移和整合素β1表达的影响。方法将Fb接种于刚度为（16．2±0．5）、（19．8±1．1）、（200．1±2．6）kPa的硅胶基底上，进行如下检测（1）分别连续培养5d或6d，进行细胞计数、噻唑蓝法检测细胞增殖活性（吸光度值表示）。（2）培养3d，检测细胞周期，计算增殖指数（PI）。（3）划痕实验后培养0（!_日）、1、2、3d，测定Fb迂移牢。（4）培养2d，流式荧光法检测细胞中整合素β1表达．对部分数据进行译因索方籍分析.结果（1）细胞计数、嚷唑蓝法榆测均显示，h增殖速度及活性均随着硅胶琏底刚度的增强而增加。细胞周期检测显永：存刚度为（16．2±0．5）、（19．8±1．1）、（200．1±2．6）kPa的硅胶琏底上，细胞的P1分别为24．8％、27．4％、32．4％。（2）培养2d，在刚度分别为（19．8±1．1）、（200．1±2．6）kPa的硅胶蔡底表面上，Fh迁移率分别为（91．4±5．1）％、（100．0±1．3）％，均明漫高于刚度为（16.2±0．5）kPa硅胶基底表面的F11迁移率[（55．8±6．8）％，F值分别为3．5、4．0，P值均小于0．01]。（3）刚度为（16．2±0．5）kPa硅胶基底表面的Fh中整合素β1，表达牢最低，仅43．2％．冈0度为（200．1±2．6）kPa硅胶尽底表两的Fb中整合素13．表达率最高（81．3％）。结论基底刚度对Fb在创两愈合和搬痕形成过程中的增殖、迁移有较大影响，这一效应与其调控Fb枢合素β1表达作用相关。Objective To study the effect of substrate stiffness on proliferation, migration of fibroblast and integrin β1 expression in fibroblast. Methods Fibroblasts were inoculated on silicon substrate with stiffness of （16.2 ±0.5）, （19.8 ±1.1）, and （200.1 ±2.6） kPa. After being cuhured for5 days or 6 days, cells were counted and cell proliferative activities （recorded as absorbance value） were assessed with methyl thiaznlyl blue （MTT）. After being cultured for 3 days, cell cycle was detected and prnliferation index （PI） was calculated. The cell scratch test was used for determination of cell migration rate on post scratch day （PSD） 0 （the day of scratch）, 1, 2, and 3. After being cultured fnr2 days, the expression ofintegrin β1 was determined by flow cytometry with fluorescence. Data were processed with one-way analysis of variance. Results （ 1 ） The proliferative speed and proliferative activity of fibroblasts were all increased along witil the increase in substrate stiffness. PI of fibroblasts inoculated on silicon substrale with stiffness of （16.2±0.5）, （19.8 ± 1. 1）, and （200. 1 ±2.6） kPa was respectively 24.8% , 27.4%, 32.4%. On PSD 2, migration rate of fibroblasts inoculated on silicon substrate with stiffness of （19. 8 ± 1. 1 ） and （200. 1 ±2.6） kPa was respectively （91.4±_5.1）%, （100.0-±1.3）%, which were higher than that offibroblasts inoculated on silicon substrate with stiffness of （ 16.2 _±0.5） kPa [ （55.8 ±6.8） % , with F value respectively 3.5, 4.0, P values all below 0.01 ]. （3） The expression rate of integrin β1 in fibroblasts inoculated on silicon substrate with stiffness of （ 16.2 ±0.5） kPa was the lowest （43.22%） , and that in fibroblast inoculated on silicon substrate with stiffness of （200. 1 ± 2.6） kPa was the highest （81.26%）. Conclusions Substrate stiffness may have a great effect on proliferation and migration of fibroblast during the process of wound healing and scar fnrmation, which can

关 键 词：成纤维细胞 细胞增殖 细胞运动 抗原 CD29 刚度 

分 类 号：R622[医药卫生—整形外科]