【作者】 雷港； 武和明； 江飞；

【Author】 LEI Gang;WU Heming;JIANG Fei;Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University;

【机构】 南京医科大学口腔疾病研究江苏省重点实验室； 南京医科大学附属口腔医院特约诊疗科； 南京医科大学附属口腔医院口腔颌面外科； 南京医科大学附属口腔医院综合诊疗科；

【摘要】 目的 :研究成釉细胞瘤细胞(human ameloblastoma-derived cells, hAMCs)对骨髓间充质干细胞(human bone marrow-derived mesenchymal stem cells, hBMSCs)增殖、成骨分化,以及自噬水平的影响,初步探讨成釉细胞瘤抑制骨髓干细胞骨再生的部分机制。方法 :组织块结合酶消化法分离培养hAMCs并鉴定,收集其条件培养液(hAMC conditional media, hAMC-CM)。以hAMC-CM刺激hBMSCs,通过CCK-8检测hBMSCs增殖变化,碱性磷酸酶实验,茜素红染色及Western blot检测hBMSCs成骨分化相关表达的变化,Western blot检测hBMSCs自噬相关指标的变化。结果 :hAMCCM处理hBMSCs后,hBMSCs形体及增殖状态没有变化,细胞早期及晚期成骨相关指标下调,同时自噬相关蛋白表达也下调。结论 :hAMCs旁分泌作用可抑制hBMSCs成骨分化,该表型与hBMSCs自噬水平下调密切相关。更多还原

【Abstract】 Objective: To explore the mechanism of ameloblastoma in inhibiting proliferation, osteogenic differentiation and autophagy level of human bone marrow derived mesenchymal stem cells(hBMSCs) to affect bone regeneration. Methods: Human ameloblastoma cells(hAMCs) were isolated and identified. The conditional media was collected to culture the hBMSCs. CCK-8 kit was used to detect the hBMSC proliferation, while ALP staining, Alizarin Red staining and Western blot were applied to investigate the osteogenic differentiation and autophagy status of the hBMSCs. Results: The osteogenic phenotype of hBMSCs were inhibited by hAMC-CM. The markers of osteogenesis and autophagy in hBMSCs were down-regulated, such as RUNX2, OSX, ALP, OCN, OPN, Collagen I, ATG5, Beclin1 and LC3 b. Conclusions: hAMCs could inhibit hBMSCs osteo-differentiation via their paracrine functions. The mechanisms of the phenotype might be associated with impacting autophagy level of hBMSCs.更多还原