血小板衍生生长因子-BB促进成骨诱导分化的适宜浓度的研究  

Appropriate concentration of platelet-derived growth factor BB in promoting osteogenic differentiation

作　　者：段东明 刘佳敏 肖天华 黎源鑫 曾裕威 魏建国 娄爱菊[2] 张凤朝 周蕾[1] 王簕[1] Duan Dongming;Liu Jiamin;Xiao Tianhua;Li Yuanxin;Zeng Yuwei;Wei Jianguo;Lou Aiju;Zhang Fengchao;Zhou Lei;Wang Le(Department of Orthopedics,the Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150,China;Rheumatism Department,Liwan Hospital Affiliated to Guangzhou Medical University,Guangzhou 510150,China;Department of Gastroenterology,Baoshan People’s Hospital of Yunnan Province,Baoshan 678099,China)

机构地区：[1]广州医科大学附属第三医院骨科,广州510150 [2]广州医科大学附属荔湾医院风湿科,广州510150 [3]云南省保山市人民医院消化内科,保山678099

出　　处：《中华实验外科杂志》2023年第2期321-324,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82072409)。

摘　　要：目的探索血小板衍生生长因子-BB(PDGF-BB)促进成骨诱导分化的适宜浓度。方法以骨形态蛋白-2(BMP-2)为阳性对照,将其与PDGF-BB按体系浓度分为7组,0μg/ml control组、0.02μg/ml PDGF组、0.20μg/ml PDGF组、2.00μg/ml PDGF组、0.02μg/ml BMP-2组、0.20μg/ml BMP-2组和2.00μg/ml BMP-2组。分别在以上浓度体系中培养,以评估细胞增殖及成骨分化。组间比较采用方差分析。结果使用慢病毒携带红色荧光蛋白(lenti-RFP)转染的骨髓间充质干细胞(BMSCs)具有良好的细胞活性,且能够正常增殖。荧光显微镜结果显示0.02μg/ml BMP-2及0.20μg/ml PDGF-BB对BMSCs具有最佳的促进增殖作用,其中0.02μg/ml BMP-2组的荧光强度高于control组、0.20μg/ml BMP-2组及2.00μg/ml组(75.18±0.37比52.69±0.69、58.19±0.63、44.95±1.01,F=651.600,P<0.05);0.20μg/ml PDGF组的荧光强度高于control组、0.02μg/ml PDGF组及2.00μg/ml PDGF组(94.39±2.62比52.69±0.69、81.82±1.18、54.78±0.69,F=365.800,P<0.05)。细胞计数试剂盒(CCK-8)结果显示0.02μg/ml BMP-2及0.20μg/ml PDGF-BB对BMSCs具有最佳的促进增殖作用,其中0.02μg/ml BMP-2组的吸光度(A)值高于control组、0.20μg/ml BMP-2组及2.00μg/ml组(0.26±0.01比0.22±0.00、0.19±0.01、0.17±0.00,F=70.440,P<0.05);0.20μg/ml PDGF组的A值高于control组、0.02μg/ml PDGF组及2.00μg/ml PDGF组(0.42±0.00比0.22±0.00、0.35±0.01、0.27±0.00,F=154.300,P<0.05)。以0.02μg/ml BMP-2培养体系作为阳性对照,茜素红S显示0.20μg/ml PDGF-BB对BMSCs具有最佳的促进成骨诱导作用,0.20μg/ml PDGF-BB组的A值高于control组、0.02μg/ml BMP-2组、0.02μg/ml PDGF-BB组及2.00μg/ml PDGF-BB组(1.88±0.07比1.10±0.04、1.47±0.05、0.90±0.03、0.65±0.01,F=227.700,P<0.05)。碱性磷酸酶试验结果显示0.20μg/ml PDGF-BB对BMSCs具有最佳的促进成骨诱导作用,0.20μg/ml PDGF-BB组的A值高于control组、0.02μg/ml BMP-2组、0.02μg/ml PDGF-BB组及2.00μg/ml PDGF-BB组(2.34±0.13比0.60±0.01、1.23±0.05Objective To explore the appropriate concentration of platelet derived growth factor-BB(PDGF-BB)to promote osteogenic-induced differentiation.Methods Bone morphogenetic protein-2(BMP-2)was used as a positive control and grouped with the PDGF-BB by systemic concentrations:0μg/ml(control),0.02μg/ml PDGF,0.20μg/ml PDGF,2.00μg/ml PDGF,0.02μg/ml BMP-2,0.20μg/ml BMP-2,and 2.00μg/ml BMP-2.Bone marrow mesenchymal stem cells(BMSCS)were cultured in the above concentration system to assess cell proliferation and osteogenic differentiation.Comparisons between groups were performed by ANOVA.Results BMSCs transfected with lentivirus carrying red fluorescent protein(lenti-RFP)had good cell activity and could proliferate normally.Fluorescence microscope photos showed that 0.02μg/ml BMP-2 and 0.20μg/ml PDGF-BB had the best effect on the proliferation of BMSCs.The fluorescence intensity in 0.02μg/ml BMP-2 group was higher than in the control group,0.20μg/ml BMP-2 group,and 2.00μg/ml BMP-2 group(75.18±0.37 vs.52.69±0.69,58.19±0.63,44.95±1.01,F=651.600,P<0.05).The fluorescence intensity in 0.20μg/ml PDGF group was higher than the control,0.02μg/ml PDGF,and 2.00μg/ml PDGF groups(94.39±2.62 vs.52.69±0.69,81.82±1.18,54.78±0.69,F=365.800,P<0.05).Cell counting kit-8(CCK-8)showed that 0.02μg/ml BMP-2 and 0.20μg/ml PDGF-BB could optimally promote the proliferation of BMSCs.The absorbance value in 0.02μg/ml BMP-2 group was higher than the control group,0.20μg/ml BMP-2 group,and 2.00μg/ml group(0.26±0.01 vs.0.22±0.00,0.19±0.01,0.17±0.00,F=70.440,P<0.05).The absorbance value in 0.20μg/ml PDGF group was higher than the control,0.02μg/ml PDGF,and 2.00μg/ml PDGF groups(0.42±0.00 vs.0.22±0.00,0.35±0.01,0.27±0.00,F=154.300,P<0.05).With 0.02μg/ml BMP-2 culture system as positive control,Alizarin Red S showed that 0.20μg/ml PDGF-BB had the best osteogenic induction effect on BMSCs.The absorbance value in 0.20μg/ml PDGF-BB group was higher than control,0.02μg/ml BMP-2,0.02μg/ml PDGF-BB and 2.00μg/ml PDGF-B

关 键 词：生长因子 细胞 成骨诱导 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]