炎症环境下维生素 D 对人牙周膜基质细胞成骨分化的影响。
Effect of vitamin D on the osteogenic differentiation of human periodontal ligament stromal cells under inflammatory conditions.
机构信息
Division of Conservative Dentistry and Periodontology, University Clinic of Dentistry, Medical University of Vienna, Vienna, Austria.
Division of Orthodontics, University Clinic of Dentistry, Medical University of Vienna, Vienna, Austria.
出版信息
J Periodontal Res. 2021 Jun;56(3):579-588. doi: 10.1111/jre.12858. Epub 2021 Feb 5.
OBJECTIVES
Vitamin D is known to activate osteogenic differentiation of human periodontal ligament stromal cells (hPDLSCs). Recently, inflammatory stimuli were shown to inhibit the transcriptional activity of hPDLSCs, but their effect on vitamin D -induced osteogenic differentiation is not known. The present study aimed to investigate whether the effects of 1,25-dihydroxvitamin D (1,25(OH) D ) and 25-hydroxvitamin D (25(OH)D ) on the osteogenic differentiation of hPDLSCs are also altered under inflammatory conditions. Furthermore, the expression of osteogenesis-related factors by hPDLSCs under osteogenic conditions was assessed in the presence of inflammatory stimuli.
MATERIALS AND METHODS
Primary hPDLSCs of six donors were cultured in osteogenic induction medium containing either 1,25(OH) D (0-10 nM) or 25(OH)D (0-100 nM) in the presence and absence of Porphyromonas gingivalis lipopolysaccharide (LPS) or Pam3CSK4 for 7, 14 and 21 days. Osteogenic differentiation of hPDLSCs was evaluated by analysis of mineralization as assessed by Alizarin Red S staining and gene expression levels of osteogenesis-related factors osteocalcin, osteopontin and runt-related transcription factor 2 (RUNX2) were analysed with qPCR.
RESULTS
Treatment with 1,25(OH) D significantly enhanced the osteogenic differentiation of hPDLSCs and their expression of osteocalcin and osteopontin. The 1,25(OH) D -triggered expression of osteogenesis-related factors was significantly lower in the presence of Pam3CSK4, but not P. gingivalis LPS. None of the inflammatory stimuli had significant effects on the 1,25(OH) D -induced osteogenic differentiation. 25(OH)D neither affected gene expression levels nor osteogenic differentiation of hPDLSCs cultured in osteogenic induction medium.
CONCLUSION
The results of this study indicate that inflammatory stimuli also diminish the 1,25(OH) D -induced expression of osteogenesis-related factors in hPDLSCs under osteogenic conditions, while having no effect on the osteogenic differentiation.
目的
维生素 D 已知能激活人牙周膜基质细胞（hPDLSCs）的成骨分化。最近，炎症刺激物被证明会抑制 hPDLSCs 的转录活性，但它们对维生素 D 诱导的成骨分化的影响尚不清楚。本研究旨在探讨在炎症条件下，1,25-二羟维生素 D（1,25（OH）D）和 25-羟维生素 D（25（OH）D）对 hPDLSCs 成骨分化的影响是否也会发生改变。此外，在存在炎症刺激物的情况下，评估 hPDLSCs 在成骨条件下成骨相关因子的表达情况。
材料和方法
从六位供体中培养的原代 hPDLSCs 在含有 1,25（OH）D（0-10 nM）或 25（OH）D（0-100 nM）的成骨诱导培养基中培养，同时存在或不存在牙龈卟啉单胞菌脂多糖（LPS）或 Pam3CSK4，7、14 和 21 天。通过茜素红 S 染色评估矿化来评估 hPDLSCs 的成骨分化，并用 qPCR 分析成骨相关因子骨钙素、骨桥蛋白和 runt 相关转录因子 2（RUNX2）的基因表达水平。
结果
1,25（OH）D 的治疗显著增强了 hPDLSCs 的成骨分化及其骨钙素和骨桥蛋白的表达。在 Pam3CSK4 存在的情况下，1,25（OH）D 触发的成骨相关因子的表达明显降低，但牙龈卟啉单胞菌 LPS 则没有。炎症刺激物均未对 1,25（OH）D 诱导的成骨分化产生显著影响。25（OH）D 既不影响成骨诱导培养基中 hPDLSCs 的基因表达水平，也不影响其成骨分化。
结论
本研究结果表明，在成骨条件下，炎症刺激物也会降低 1,25（OH）D 诱导的 hPDLSCs 中成骨相关因子的表达，而对成骨分化没有影响。
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