转化生长因子-β_3对兔牙髓干细胞增殖及骨向分化的影响  

Influences of transforming growth factor-β_3 on proliferation and osteogenic differentiation of rabbit dental pulp stem cells

作　　者：刘晓文[1] 木合塔尔.霍加 麦麦提依明.哈力克 帕尔哈提.阿布肚热合曼 庄友梅[1] 张晓莉[1] 买布拜木.买买提依明 

机构地区：[1]新疆维吾尔自治区人民医院口腔科,乌鲁木齐830001

出　　处：《中华实用诊断与治疗杂志》2016年第3期242-245,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：国家自然科学基金(81560180)

摘　　要：目的探讨转化生长因子-β3（transforminggrowthfactor-β3，TGF-β3）对体外培养的兔牙髓干细胞（dentalpulpstemceils，DPSCs）增殖和分化的影响。方法采用酶解组织块法将兔DPSCs分离培养获得DPSCs，光镜下行形态学观察；将体外培养的第3代DPSCs分为5组，分别为对照组、20μg／LTGF-β3组、40μg／LTGF-β3组、80tag／LTGF-β3组和100μg／LTGF-β3组，分别加入0、20、40、80、100μg／LTGF-β3；采用MTT法检测5组DPSCsA490值，采用免疫细胞化学法检测成骨细胞标记物骨钙素（osteocalcin，OC）、I型胶原酶（collagenI，Col—I）、骨涎蛋白（bonesialoprotein，BSP）表达情况，采用茜素红染色检测出现矿化结节情况，并进行比较。结果兔DPSCs呈集落状生长，在体外有一定克隆形成能力；对照组及20、40、80、100μg／LTGF-β3组兔DPSCsA490值分别为0．34±0．55、0．34±0．19、0．33±0．47、0．334±0．30、0．34±0．47，各组间两两比较差异均无统计学意义（P〉0．05）；80、100μg／LTGF—B3组诱导后第3天出现Col—I阳性表达，第7天开始消失，第5-7天出现BSP阳性表达，第7-14天出现0c阳性表达，第7天出现矿化结节；40μg／LTGF-β3陆组第5-7天均有Col—I阳性表达，第7天出现BSP阳性表达，第14天出现OC阳性表达，第14天出现矿化结节；20μg／L组第7天出现Col—I阳性表达，第14天消失，第14天出现BSP阳性表达，第21天出现OC阳性表达，第21天出现矿化结节；对照组Col-I、BSP和OC均为阴性，未出现矿化结节。结论TGF-β3风对体外培养的兔DPSCs增殖无影响，但对兔DPSCs向成骨细胞分化能力有一定促进作用，并在一定范围内呈浓度依赖关系。Objective To investigate the influence of transforming growth factor-β3 （TGF-a） on the proliferation and differentiation of rabbit dental pulp stem cells （DPSCs）. Methods DPSCs were isolated with tissue enzymetic digestion method to observe the morphology under light microscope. The third-passage DPSCs cultured in vitro were divided into five groups, incubated with 0 vg/L （control group）, 20μg/L TGF-β3（20 μg/L group）, 40 μg/L TGF-β3 （40 μg/L group）, 80 μg/L TGF-β3（μg/L group）, and 100 μg/L TGF--β3 （100 μg/L group）, respectively. MTT assay was adopted to detect the A490 values in five groups. Immunocytochemistry staining was adopted to detect the expressions of osteocalcin, collagen I （Col-I ） and bone sialoprotein （BSP）. Alizarin red staining was adopted to detect mineralized nodules. Results DPSCs in rabbits grew in clones, having cloning ability in vitro. A490 values were 0.34±0.55, 0.34± 0.19, 0.33±0.47, 0.33±0.30 and 0.34±0.47 respectively in control group, and 20, 40, 80 and 100 μg/L groups, showing no significant differences between each two groups （P〈0.05）. In 80 and 100 μg/L groups, Col-I was positively expressed by day 3 and began to disappear by day 7, BSP was positively expressed by day 5 to 7, osteocalcin was positively expressed by day 7 to 14, and mineralized nodules were found by day 7 after induction. In 40 gg/L group, Col-I was positively expressed by day 5 to 7, BSP was positively expressed by day 7, osteocalcin was positively expressed by day 14 and mineralized nodules were found by day 14 after induction. In 20 gg/L group, Col- I was positively expressed by day 7 and began to disappear by day 14. BSP was positively expressed by day 14, osteocalcin was positively expressed by day 21, and mineralized nodules were found by day 21 after induction. Col-I, BSP and osteocalcin were negatively expressed and no mineralized nodule was found in control group. Conclusion TGF-β3 accelerates the osteogenic differentiation of DPSCs in a dose depe

关 键 词：牙髓干细胞 转化生长因子-Β3 增殖 骨分化 

分 类 号：R781[医药卫生—口腔医学]