TLR2 和 TLR4 对人牙周膜成纤维细胞成骨能力的调节作用存在差异。

TLR2 and TLR4 Differentially Regulate the Osteogenic Capacity of Human Periodontal Ligament Fibroblasts.

机构信息

Molecular Medicine Program, Faculty of Science, Mahidol University, Ratchthewi, Bangkok Thailand; Department of Applied Oral Sciences, The Forsyth Institute, Cambridge, MA, USA; and Department of Oral Medicine and Periodontology, Faculty of Dentistry, Mahidol University, Ratchthewi, Bangkok Thailand.

Department of Oral Biology, Faculty of Dentistry, Mahidol University, Ratchthewi, Bangkok Thailand.

出版信息

J Int Acad Periodontol. 2021 Jan 1;23(1):3-10.

PMID:33512337

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8142849/

Abstract

AIMS

To test that the osteogenic capacity of periodontal ligament (PDL) fibroblasts can be mediated by TLR2 and TLR4 activation.

MATERIALS AND METHODS

Human PDL fibroblasts were cultured in osteogenic medium and treated with TLR2 and TLR4 agonists (Pam3CSK4 and monophosphoryl Lipid A (MPLA), respectively). Cell proliferation was measured by MTT and BrdU incorporation. Osteogenic differentiation was measured by alkaline phosphatase (ALP) activity. Nodule formation was measured for osteoblast function. The expression of markers of potential signaling pathways (RUNX2, OCN, BSP and Osterix) was evaluated by quantitative PCR.

RESULTS

PDL fibroblasts grew at the same rate during the first 5 days in response to both Pam3CSK5 and MPLA. On day 7, cells cultured in the presence of Pam3CSK4 had a significantly higher rate of DNA replication, while cells in MPLA group had a significantly lower DNA replication rate (one-third) compared to the control (p less than 0.05). Pam3CSK4 induced significantly higher ALP activity and larger calcified nodules. TLR4 activation significantly reduced the expression of RUNX2 and osterix and enhanced OCN. Neither TLR2 nor TLR4 affected BSP expression.

CONCLUSIONS

These data suggest that the activation of TLR2 and TLR4 differentially and perhaps antagonistically modulate osteogenesis by human PDL fibroblasts and have a direct role of TLR-mediated PDL function during periodontal regeneration as a potential target for therapeutics.

摘要

目的

检测 TLR2 和 TLR4 的激活是否能调节牙周膜（PDL）成纤维细胞的成骨能力。

材料与方法

将人牙周膜成纤维细胞培养在成骨培养基中，并分别用 TLR2 和 TLR4 激动剂（Pam3CSK4 和单磷酰脂质 A（MPLA））处理。通过 MTT 和 BrdU 掺入法测量细胞增殖。通过碱性磷酸酶（ALP）活性测量成骨分化。通过矿化结节形成测量成骨细胞功能。通过定量 PCR 评估潜在信号通路（RUNX2、OCN、BSP 和 Osterix）标志物的表达。

结果

在 Pam3CSK5 和 MPLA 作用下，PDL 成纤维细胞在前 5 天的生长速度相同。第 7 天，在 Pam3CSK4 存在下培养的细胞的 DNA 复制率显著更高，而 MPLA 组的细胞的 DNA 复制率（三分之一）显著低于对照组（p<0.05）。Pam3CSK4 诱导的 ALP 活性更高，矿化结节更大。TLR4 激活显著降低了 RUNX2 和 Osterix 的表达，增强了 OCN。TLR2 和 TLR4 均不影响 BSP 的表达。