Gingival fibroblasts produce paracrine signals that affect osteoclastogenesis in vitro

IF 2.1 Q3 ENDOCRINOLOGY & METABOLISM
Solen Novello , Ton Schoenmaker , Teun J. de Vries , Behrouz Zandieh Doulabi , Astrid D. Bakker , Marja L. Laine , Ineke D.C. Jansen
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Abstract

In periodontitis, gingival fibroblasts (GF) appear to produce a multitude of paracrine factors. However, the influence of GF-derived soluble factors on osteoclastogenesis remains unclear. In this case study, production of paracrine factors by GF was assessed under inflammatory and non-inflammatory conditions, as well as their effect on osteoclastogenesis. Human primary GF were cultured in a transwell system and primed with a cocktail of IL-1β, IL-6 and TNF-α to mimic inflammation. GF were co-cultured directly and indirectly with human peripheral blood mononuclear cells (PBMC). Cytokines and chemokines in supernatants (flow cytometry based multiplex assay), osteoclastogenesis (TRAcP staining) and gene expression (qPCR) were quantified on days 7 and 21.

Results from this case study showed that GF communicated via soluble factors with PBMC resulting in a two-fold induction of osteoclasts. Reversely, PBMC induced gene expression of IL-6, OPG and MCP-1 by GF. Remarkably, after priming of GF with cytokines, this communication was impaired and resulted in fewer osteoclasts. This could be partly explained by an increase in IL-10 expression and a decrease in MCP-1 expression. Intriguingly, the short priming of GF resulted in significantly higher expression of inflammatory cytokines that was sustained at both 7 and 21 days.

GF appear to produce paracrine factors capable of stimulating osteoclastogenesis in the absence of physical cell-cell interactions. GF cultured in the presence of PBMC or osteoclasts had a remarkably inflammatory phenotype. Given profound expression of both pro- and anti-inflammatory cytokines after the inflammatory stimulus, it is probably the effector hierarchy that leads to fewer osteoclasts.

牙龈成纤维细胞产生影响体外破骨细胞生成的旁分泌信号
在牙周炎中,牙龈成纤维细胞(GF)似乎会产生多种旁分泌因子。然而,GF衍生的可溶性因子对破骨细胞生成的影响仍不清楚。本病例研究评估了在炎症和非炎症条件下牙龈成纤维细胞产生的旁分泌因子及其对破骨细胞生成的影响。将人类原代 GF 培养在一个 transwell 系统中,并用 IL-1β、IL-6 和 TNF-α 鸡尾酒诱导以模拟炎症。GF 直接或间接与人外周血单核细胞(PBMC)共培养。第 7 天和第 21 天,对上清液中的细胞因子和趋化因子(基于流式细胞仪的多重检测)、破骨细胞生成(TRAcP 染色)和基因表达(qPCR)进行量化。相反,PBMC 通过 GF 诱导 IL-6、OPG 和 MCP-1 的基因表达。值得注意的是,在用细胞因子引诱 GF 后,这种沟通受到了影响,导致破骨细胞数量减少。部分原因可能是 IL-10 表达的增加和 MCP-1 表达的减少。耐人寻味的是,GF 的短时间引物导致炎性细胞因子的表达显著增加,并在 7 天和 21 天内持续。在有 PBMC 或破骨细胞存在的情况下培养的 GF 具有明显的炎症表型。鉴于炎症刺激后促炎症细胞因子和抗炎症细胞因子都会大量表达,可能是效应层次导致破骨细胞减少。
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来源期刊
Bone Reports
Bone Reports Medicine-Orthopedics and Sports Medicine
CiteScore
4.30
自引率
4.00%
发文量
444
审稿时长
57 days
期刊介绍: Bone Reports is an interdisciplinary forum for the rapid publication of Original Research Articles and Case Reports across basic, translational and clinical aspects of bone and mineral metabolism. The journal publishes papers that are scientifically sound, with the peer review process focused principally on verifying sound methodologies, and correct data analysis and interpretation. We welcome studies either replicating or failing to replicate a previous study, and null findings. We fulfil a critical and current need to enhance research by publishing reproducibility studies and null findings.
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