中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2011年
6期
427-431
,共5页
王玉%王贵学%罗向东%邱菊辉
王玉%王貴學%囉嚮東%邱菊輝
왕옥%왕귀학%라향동%구국휘
成纤维细胞%细胞增殖%细胞运动%抗原%CD29%刚度
成纖維細胞%細胞增殖%細胞運動%抗原%CD29%剛度
성섬유세포%세포증식%세포운동%항원%CD29%강도
Fibroblasts%Cell proliferation%Cell movement%Antigens,CD29%Stiffness
目的 了解基底刚度对Fb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5 d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(当日)、1、2、3 d,测定Fb迁移率.(4)培养2 d,流式荧光法检测细胞中整合素β1表达.对部分数据进行单因素方差分析. 结果 (1)细胞计数、噻唑蓝法检测均显示,Fb增殖速度及活性均随着硅胶基底刚度的增强而增加.细胞周期检测显示:在刚度为(16.2±0.5)、(19.8±1.1)(200.1±2.6)kPa的硅胶基底上,细胞的PI分别为24.8％、27.4％、32.4％.(2)培养2d,在刚度分别为(19.8±1)、(200.1±2.6) kPa的硅胶基底表面上,Fb迁移率分别为(91.4±5.1)％(100.0±1.3)％,均明显高于刚度为(16.2±0.5)kPa硅胶基底表面的Fb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Fb中整合素β1表达率最低,仅43.2％；刚度为(200.1±2.6)kPa硅胶基底表面的Fb中整合素β1表达率最高(81.3％]. 结论 基底刚度对Fb在创面愈合和瘢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整合素β1表达作用相关.
目的 瞭解基底剛度對Fb增殖、遷移和整閤素β1錶達的影響. 方法 將Fb接種于剛度為(16.2±0.5)、(19.8±1.1)、(200.1±2.6)kPa的硅膠基底上.進行如下檢測.(1)分彆連續培養5 d或6d,進行細胞計數、噻唑藍法檢測細胞增殖活性(吸光度值錶示).(2)培養3d,檢測細胞週期,計算增殖指數(PI).(3)劃痕實驗後培養0(噹日)、1、2、3 d,測定Fb遷移率.(4)培養2 d,流式熒光法檢測細胞中整閤素β1錶達.對部分數據進行單因素方差分析. 結果 (1)細胞計數、噻唑藍法檢測均顯示,Fb增殖速度及活性均隨著硅膠基底剛度的增彊而增加.細胞週期檢測顯示:在剛度為(16.2±0.5)、(19.8±1.1)(200.1±2.6)kPa的硅膠基底上,細胞的PI分彆為24.8％、27.4％、32.4％.(2)培養2d,在剛度分彆為(19.8±1)、(200.1±2.6) kPa的硅膠基底錶麵上,Fb遷移率分彆為(91.4±5.1)％(100.0±1.3)％,均明顯高于剛度為(16.2±0.5)kPa硅膠基底錶麵的Fb遷移率[(55.8±6.8)％,F值分彆為3.5、4.0,P值均小于0.01].(3)剛度為(16.2±0.5)kPa硅膠基底錶麵的Fb中整閤素β1錶達率最低,僅43.2％；剛度為(200.1±2.6)kPa硅膠基底錶麵的Fb中整閤素β1錶達率最高(81.3％]. 結論 基底剛度對Fb在創麵愈閤和瘢痕形成過程中的增殖遷移有較大影響,這一效應與其調控Fb整閤素β1錶達作用相關.
목적 료해기저강도대Fb증식、천이화정합소β1표체적영향. 방법 장Fb접충우강도위(16.2±0.5)、(19.8±1.1)、(200.1±2.6)kPa적규효기저상.진행여하검측.(1)분별련속배양5 d혹6d,진행세포계수、새서람법검측세포증식활성(흡광도치표시).(2)배양3d,검측세포주기,계산증식지수(PI).(3)화흔실험후배양0(당일)、1、2、3 d,측정Fb천이솔.(4)배양2 d,류식형광법검측세포중정합소β1표체.대부분수거진행단인소방차분석. 결과 (1)세포계수、새서람법검측균현시,Fb증식속도급활성균수착규효기저강도적증강이증가.세포주기검측현시:재강도위(16.2±0.5)、(19.8±1.1)(200.1±2.6)kPa적규효기저상,세포적PI분별위24.8％、27.4％、32.4％.(2)배양2d,재강도분별위(19.8±1)、(200.1±2.6) kPa적규효기저표면상,Fb천이솔분별위(91.4±5.1)％(100.0±1.3)％,균명현고우강도위(16.2±0.5)kPa규효기저표면적Fb천이솔[(55.8±6.8)％,F치분별위3.5、4.0,P치균소우0.01].(3)강도위(16.2±0.5)kPa규효기저표면적Fb중정합소β1표체솔최저,부43.2％；강도위(200.1±2.6)kPa규효기저표면적Fb중정합소β1표체솔최고(81.3％]. 결론 기저강도대Fb재창면유합화반흔형성과정중적증식천이유교대영향,저일효응여기조공Fb정합소β1표체작용상관.
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 cultured for 5 days or 6 days,cells were counted and cell proliferative activities ( recorded as absorbance value) were assessed with methyl thiazolyl blue (MTT).After being cultured for 3 days.cell cycle was detected and proliferation 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 for 2 days,the expression of integrin β1 was determined by flow cytometry with fluorescence.Data were processed with one-way analysis of variance,Results (1)The proliferaive speed and proliferative activity of fibroblasts were all increased along with the increase in substrate stiffness.PI of fibroblasts inoculated on silicon substrate 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 of fibroblasts 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 substrata with stiffness of ( 200.1 ± 2.6 ) kPa was the highest ( 81.26％ ).Conclusions Substrate stiffnese may have a great effect on proliferation and migration of fibroblast during the process of wound healing and scar formation,which can be related to regulation of intergrin β1 expression.
