槲皮素对牙髓间充质细胞成骨能力及兔颅骨缺损修复的影响  

Effect of quercetin on osteogenic ability of dental pulp mesenchymal cells and repair of calvarial defects in rabbits

作　　者：胡韶光 刘珂珂 汪芹芹 韩旭 桂双英 徐燕[1] HU Shao-guang;LIU Ke-ke;WANG Qin-qin;HAN Xu;GUI Shuang-ying;XU Yan(School of Stomatology, Anhui Medical University, Affiliated Stomatological Hospital of Anhui Medical University, Laboratory of Anhui Oral Disease Research Center, Hefei 230032, China;School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China)

机构地区：[1]安徽医科大学口腔医学院,安徽医科大学附属口腔医院,安徽省口腔疾病研究中心实验室,安徽合肥230032 [2]安徽中医药大学药学院,安徽合肥230012

出　　处：《中国药理学通报》2021年第11期1536-1541,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No81873019);安徽高校自然科学研究项目(NoKJ2017ZD17)。

摘　　要：目的研究槲皮素对人牙髓间充质细胞(human dental pulp mesenchymal stromal cells,hDPSCs)体外及体内成骨能力的影响。方法通过对牙髓组织培养获得hDPSCs,流式鉴定特征性表面抗原;CCK-8检测细胞毒性;碱性磷酸酶(ALP)和茜素红染色检测细胞体外成骨能力;qPCR检测成骨基因的表达;在10只雄性新西兰兔颅骨上造4个8 mm直径的圆形骨缺损,随机分为4组:A组含1μmol·L-1槲皮素的完全培养液培养的hDPSCs接种于Bio-Oss骨粉组,B组常规完全培养液培养的hDPSCs接种于Bio-Oss骨粉组,C组Bio-Oss组,D组空白组。术后4、8周取标本行Micro-CT三维重建,计算骨体积分数评估成骨效果。结果低浓度的槲皮素无细胞毒性且1μmol·L-1可促进细胞增殖;0.1、1μmol·L-1槲皮素可提高ALP的活性水平;ALP和茜素红染色结果进一步证实槲皮素后能够促进ALP和钙结节的形成;Micro-CT显示在术后4、8周A组新骨形成较B、C组多,D组成骨效果不明显。结论槲皮素可促进hDPSCs的增殖与体外及体内的成骨分化。Aim To investigate the effect of quercetin on the osteogenic ability of human dental pulp mesenchymal stromal cells(hDPSCs)in vitro and in vivo.Methods hDPSCs were obtained from the pulp tissues of premolar,and the characteristic surface antigens were identified by flow cytometry.Cell Counting Kit-8(CCK-8)assay was used to test the cytotoxicity of quercetin.Alkaline phosphatase(ALP)and alizarin red staining were used to detect the osteogenic ability of cells in vitro.The expression of osteogenic genes was detected by qPCR.Four round calvarial bone defects with a diameter of 8mm were created in 10 male New Zealand rabbits,and they were differentiated and randomized into four groups.Group A,hDPSCs cultured on Bio-Osswith complete culture medium containing 1μmol·L-1 quercetin;Group B,hDPSCs cultured on Bio-Osswith complete culture medium;Group C,Bio-Ossonly;Group D,blank.Specimens were taken for Micro-CT three-dimensional reconstruction,and bone volume fraction was calculated to evaluate the osteogenesis effect 4 and 8 weeks after surgery.Results CCK-8 showed that quercetin had no cytotoxicity at low concentration and it could promote cell proliferation at the concentration of 1μmol·L-1.Quercetin could increase the activity level of alkaline phosphatase by quantitative evaluation of ALP activity.It was further confirmed that quercetin could promote the formation of alkaline phosphatase and calcium nodules by the results of ALP and Alizarin Red staining.Bone formation in group A was more than that in group B and group C 4 and 8 weeks after surgery,and the effect of bone formation in group D was not obvious by Micro-CT.Conclusion Quercetin can promote the proliferation and osteogenic differentiation of hDPSCs in vitro and in vivo.

关 键 词：槲皮素 间充质基质细胞 组织工程 成骨分化 颅骨缺损 骨移植 

分 类 号：R-332[医药卫生] R285.5