褪黑素对人牙周膜细胞成骨分化能力的影响

目的探讨褪黑素对人牙周膜细胞(hPDLCs)成骨分化能力的影响。方法原代培养法成功提取hPDLCs后,加入不同浓度的褪黑素作为实验组,空白对照组不加褪黑素。MTT法检测不同浓度褪黑素对细胞增殖的影响,相关试剂盒检测碱性磷酸酶(ALP)的活性和表达后,采用茜素红S染色、Real-time PCR和Western Blot分别研究褪黑素对hPDLCs的矿化结节、成骨相关mRNA以及蛋白表达的影响。结果与对照组相比,褪黑素组hPDLCs的增殖活力略有抑制,其中50μmol/L和100μmol/L褪黑素组抑制最为显著(P<0.01);褪黑素组均表现为ALP活性和表达上升,矿化结节形成增多且呈浓度依赖性(P<0.01);Real-time PCR和Western Blot结果显示褪黑素组成骨相关mRNA和蛋白表达均有上升(P<0.01)。结论褪黑素可促进hPDLCs的成骨分化,具有作为牙周再生治疗药物的潜能。

Effects of melatonin on the osteogenesis of human periodontal ligament cells

Objective To evaluate the effects of melatonin on the osteogenic differentiation of human periodontal ligament cells(hPDLCs).Methods After extracting hPDLCs through primary culture,melatonin with different concentrations was added into hPDLCs as experimental groups.The blank control group was not added with melatonin.The effects of melatonin in different concentrations on proliferation of hPDLCs were assessed by MTT assay.Relevant kits were used to assay Alkaline phosphatase(ALP)activity and expression.Alizarin red S staining,Real-time PCR and Western Blot were used respectively to observe the formation of mineral nodules,osteoblast related mRNA and protein expression of hPDLCs.Results Compared with the control group,proliferation of hPDLCs in melatonin groups decreased,especially in 50μmol/L and 100μmol/L concentration(P<0.01).Melatonin was shown to enhance ALP activity and expression of hPDLCs.More mineral nodules were observed in melatonin groups compared to the control group and nodules were concentration dependent(P<0.01).Real-time PCR and Western Blot results showed that the osteoblast related mRNA and protein expression increased in melatonin groups(P<0.01).Conclusion Melatonin has a promoting effect on the osteogenic differentiation of hPDLCs,and it has the potential to becoming periodontal regeneration treatment.