Department of Experimental Medical Science, Lund University, Lund, Sweden.
Department of Periodontology, Faculty of Odontology, Malmö University, Malmö, Sweden.
J Periodontal Res. 2015 Oct;50(5):666-73. doi: 10.1111/jre.12249. Epub 2014 Dec 12.
The aim of this study was to assess the impact of 1α,25-dihydroxyvitamin D3 (vitamin D3) on osteogenic and inflammatory properties of human periodontal ligament (PDL) cells and investigate underlying mechanisms.
Human PDL cells, obtained from four subjects, were stimulated with vitamin D3 for 4-48 h. The bone markers osteopontin and osteocalcin and proinflammatory cytokine/chemokine expression was determined by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Cytokine and chemokine expression was determined after stimulation with the inflammation promoter lipopolysaccharide (LPS) in the presence or absence of vitamin D3. Alkaline phosphatase activity was assessed using p-nitrophenylphosphate substrate.
Treatment with 30 ng/mL of vitamin D3, corresponding to an optimal plasma concentration of vitamin D, for 24 h had no effect on PDL cell number and morphology but increased PDL cell osteopontin and osteocalcin mRNA expression by about 70 and 40%, respectively, and, moreover, treatment with vitamin D3 for 48 h enhanced PDL cell alkaline phosphatase activity by about two times showing that vitamin D3 exerts pro-osteogenic effects in human PDL cells. Stimulation with LPS (1 μg/mL) for 4 h increased PDL cell interleukin (IL)-6 cytokine and chemokine ligand 1 (CXCL1) chemokine mRNA expression several fold. The LPS-induced increase in IL-6 and CXCL1 transcripts was attenuated by vitamin D3 (30 ng/mL). Treatment with vitamin D3 (3-300 ng/mL) for 24 h reduced the LPS-evoked increase in PDL cell IL-6 protein by about 50%. Vitamin D3 (30 ng/mL) had no effect on LPS-induced IL-1β and MCP-1 mRNA expression.
Vitamin D3 promotes osteogenic differentiation but also downregulates inflammation promoter-induced IL-6 cytokine and CXCL1 chemokine expression in human PDL cells, suggesting that vitamin D3 both stimulates bone regeneration and antagonizes inflammation in human periodontal tissue.
本研究旨在评估 1α，25-二羟维生素 D3（维生素 D3）对人牙周韧带（PDL）细胞成骨和炎症特性的影响，并探讨其潜在机制。
从四位供体中获取人 PDL 细胞，用维生素 D3 刺激 4-48 小时。通过实时定量聚合酶链反应和酶联免疫吸附试验测定骨标志物骨桥蛋白和骨钙素以及促炎细胞因子/趋化因子的表达。在存在或不存在维生素 D3 的情况下，用炎症启动子脂多糖（LPS）刺激后测定细胞因子和趋化因子的表达。用对硝基苯磷酸酯底物评估碱性磷酸酶活性。
用 30ng/mL 的维生素 D3（相当于最佳血浆维生素 D 浓度）处理 24 小时对 PDL 细胞数量和形态无影响，但使 PDL 细胞骨桥蛋白和骨钙素 mRNA 表达分别增加约 70%和 40%，此外，用维生素 D3 处理 48 小时使 PDL 细胞碱性磷酸酶活性增加约两倍，表明维生素 D3 对人 PDL 细胞具有促成骨作用。用 1μg/mL LPS（4 小时）刺激使 PDL 细胞白细胞介素（IL）-6 细胞因子和趋化因子配体 1（CXCL1）趋化因子 mRNA 表达增加数倍。维生素 D3（30ng/mL）减弱了 LPS 诱导的 IL-6 和 CXCL1 转录物的增加。用维生素 D3（3-300ng/mL）处理 24 小时使 LPS 诱导的 PDL 细胞 IL-6 蛋白增加减少约 50%。维生素 D3（30ng/mL）对 LPS 诱导的 IL-1β和 MCP-1 mRNA 表达无影响。
维生素 D3 促进成骨分化，但也下调 LPS 诱导的人牙周组织中 IL-6 细胞因子和 CXCL1 趋化因子表达，表明维生素 D3 既能刺激骨再生，又能拮抗人牙周组织的炎症。
