CAJ | 학술논문

目的探讨不同局部麻醉药对人脂肪间充质干细胞(ADMSCs)的凋亡及向成骨方向分化的影响.方法应用差速贴壁法对人ADMSCs原代培养,鉴定第3代ADMSCs表面抗原CD29、CD105表达,并检测其成脂和成心肌分化能力.将第3代ADMSC,细胞分为4组,3实验组分别加入终浓度为200 mg/L的利多卡因、罗哌卡因和布比卡因干预ADMSCs,对照组加入普通DMEM低糖培养基.分别应用PI-Hoechst33342和茜素红染色法检测各组ADMSCs凋亡率和成骨细胞分化率.结果经不同局麻药干预3、5、7d后,对照组的细胞凋亡率分别为(10.2±3.5)％、(12.4±3.6)％、(16.9±3.9)％,布比卡因组分别为(15.3±2.8)％、(23.6±3.8)％、(27.4±4.6)％,利多卡因组分别为(31.8±4.2)％、(42.4±5.5)％、(61.6±6.9)％,罗哌卡因分别为(34.8±5.9)％、(43.2±6.8)％、(64.9±5.8)％.利多卡因和罗哌卡因组在不同时点的细胞凋亡数量均明显高于对照组(均P＜0.05),而且也明显高于布比卡因组(均P＜0.05).结论利多卡因、罗哌卡因能增加ADMSCs细胞凋亡比例,但布比卡因可抑制ADMSC凋亡.
목적 탐토불동국부마취약대인지방간충질간세포(ADMSCs)적조망급향성골방향분화적영향.방법 응용차속첩벽법대인ADMSCs원대배양,감정제3대ADMSCs표면항원CD29、CD105표체,병검측기성지화성심기분화능력.장제3대ADMSC,세포분위4조,3실험조분별가입종농도위200 mg/L적리다잡인、라고잡인화포비잡인간예ADMSCs,대조조가입보통DMEM저당배양기.분별응용PI-Hoechst33342화천소홍염색법검측각조ADMSCs조망솔화성골세포분화솔.결과 경불동국마약간예3、5、7d후,대조조적세포조망솔분별위(10.2±3.5)％、(12.4±3.6)％、(16.9±3.9)％,포비잡인조분별위(15.3±2.8)％、(23.6±3.8)％、(27.4±4.6)％,리다잡인조분별위(31.8±4.2)％、(42.4±5.5)％、(61.6±6.9)％,라고잡인분별위(34.8±5.9)％、(43.2±6.8)％、(64.9±5.8)％.리다잡인화라고잡인조재불동시점적세포조망수량균명현고우대조조(균P＜0.05),이차야명현고우포비잡인조(균P＜0.05).결론 리다잡인、라고잡인능증가ADMSCs세포조망비례,단포비잡인가억제ADMSC조망.
Objective To investigate the effect of different anesthetic conditions of lidocaine,ropivacaine and bupivacaine on the apoptosis and osteogenic differentiation of human Adipose-derived mesenchymal stem cells (ADMSCs).Methods ADMSCs of inguinal adipose tissue were isolated and cultured primarily by differential adhesion method.The expression of CD29 and CD105 of Passage 3 (P3) ADMSCs was detected by immunofluorescence,and the myocardium and osteogenic differentiation potential were detected.P3 ADMSCs were treated with 200 μg/mL of final concentration medium which contained lidocaine,ropivacaine and bupivacaine.Apoptosis,osteogenic and ADMSCs were detected by PI-Hoechst33342 double staining and alizarin red.Results The apoptotic proportinn (AP) of ADMSCs at Day 3,5,and 7 in control group was (10.2 ± 3.5) ％,(12.4 ± 3.6) ％ and (16.9±3.9) ％; AP in bupivacaine group was (15.3±2.8) ％,(23.6±3.8) ％ and (27.4±4.6) ％; AP in lidocaine group was (31.8 ±4.2) ％,(42.4 ±5.5) ％,and (61.6 ±6.9) ％ ; AP in ropivacaine group was (34.8 ± 5.9) ％,(43.2 ± 6.8) ％ and (64.9 ± 5.8) ％.Moreover,the AP of ADMSCs was significantly higher in lidocaine and ropivacaine group than in control group at all-time points (all P ＜ 0.05),and it was also higher than in bupivacaine group (all P ＜ 0.05).Conclusions With the culture time of ADMSCs extended by the intervention of lidocaine,ropivacaine and bupivacaine,the apoptotic proportion of ADMSCs is increasing correspondingly.But it has no influence on the osteogenic differentiation of ADMSCs.