OPG过表达的ADSC对大鼠牙槽后槽骨缺损的修复作用  

The Osteogenesis of ADSC with OPG Overexpression Has A Great Effect on the Posterior Alveolar Bone Defect

作　　者：尹鹏 季于琪 YIN Peng;JI Yuqi(Department of Stomatology,Leshan People's Hospital,Leshan 614000,Sichuan,China)

机构地区：[1]乐山市人民医院口腔科,四川乐山614000

出　　处：《中国美容医学》2024年第3期17-21,25,共6页Chinese Journal of Aesthetic Medicine

摘　　要：目的:探究骨保护素(Osteoprotegerin,OPG)过表达的脂肪间充质干细胞(Adipose-derived stem cell,ADSC)对大鼠牙槽后槽骨缺损修复的影响。方法:选择SD雄性大鼠38只,其中6只用于获得ADSC及ADSCOPG过表达转染,鉴定转染情况并观察不同ADSC的形态、成骨分化、成脂分化及增殖能力;其中8只作为对照组,另外24只建立牙槽后槽骨缺损大鼠模型,随机分为模型组、正常ADSC组和OPG过表达的ADSC组。6周后,Micro-CT扫描观察骨缺损愈合情况;苏木素-伊红(Hematoxylin-eosin,HE)染色观察骨缺损组织病理改变;免疫组化分析组织中碱性磷酸酶(Alkaline phosphatase,ALP)、骨钙素(Osteocalcin,OCN)的表达;Westernblot分析组织中OPG/核因子-κB受体活化因子配体(Receptor activator of nuclearfactor-κB ligand,RANKL)/核因子-κB受体活化因子(Receptor activator of nuclear factor-κB,RANK)通路蛋白表达。结果:携带空载体与OPG过表达慢病毒的转染效率分别为75.23%和79.08%;两种ADSC均为长梭形;OPG过表达ADSC中的OPG蛋白表达水平、增殖能力、成骨能力明显高于正常ADSC,成脂能力明显低于正常ADSC(P<0.05);与对照组相比,模型组的牙槽后槽骨缺损体积、RANKL、RANK蛋白表达水平明显升高,Lane-Sandhu组织学评分、ALP、OCN阳性表达率、OPG蛋白表达水平明显降低(P<0.05);与模型组相比,正常ADSC组获得与上述相反的结果;OPG过表达的ADSC组上述指标的变化幅度较正常ADSC组更显著。结论:OPG过表达的ADSC能够促进大鼠牙槽后槽骨缺损的修复。Objective To investigate the effect of adipose-derived stem cell(ADSC) overexpressed with osteoprotegerin(OPG)on the repair of posterior alveolar bone defects in rats. Methods Thirty-eight male SD rats were selected, of which 6 were used to obtain the overexpression of ADSC and ADSC OPG. The transfection was identified and the morphology,osteogenic differentiation,adipogenic differentiation and proliferation of different ADSC were observed. Among them, 8 rats were used as control group, and 24 rats with alveolar bone defect were randomly divided into model group, normal ADSC group and OPG overexpression ADSC group. Six weeks later, Micro-CT scan was used to observe the healing of bone defect in rats.The histopathological changes of bone defects were observed by hematoxylin-eosin(HE) staining. The expression of alkaline phosphatase(ALP) and osteocalcin(OCN) in the tissue was analyzed by immunohistochemistry. The expression of OPG/receptor activator of nuclear factor-κB ligand(RANKL)/Receptor activator of nuclear factor-κB(RANK) pathway was analyzed by Western blot. Results The transfection efficiency of empty vector and OPG overexpression lentivirus was 75.23%and 79.08%, respectively. Both kinds of ADSC were fusiform. The expression level of OPG protein, proliferation ability and osteogenic ability in OPG overexpressed ADSC were significantly higher than those in normal ADSC, while the adipogenic ability was significantly lower than that in normal ADSC(P<0.05). Compared with the control group, the volume of posterior alveolar bone defect, the expression of RANKL and RANK protein in the model group increased significantly, while the histological score of Lane-Sandhu, the positive expression rate of ALP and OCN and the expression level of OPG protein decreased significantly in the model group(P<0.05). Compared with the model group, the normal ADSC group achieved the opposite results. The changes of the above indexes in the ADSC group with overexpression of OPG were more significant than those in the normal ADSC gr

关 键 词：脂肪间充质干细胞 骨保护素 牙槽后槽骨缺损 大鼠 成脂分化能力 成骨分化能力 

分 类 号：R781.42[医药卫生—口腔医学]