Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.
Department of Surgery, Stomatology, Pathology and Radiology, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.
BMC Immunol. 2020 Jun 22;21(1):38. doi: 10.1186/s12865-020-00367-8.
Oral fibroblast immunological responses to bacterial stimuli are well known. However, there are few studies about pulp fibroblasts from deciduous teeth (HDPF) responses, which are important for the treatment of pulp infections in children. The aim of this study was to evaluate expression and production of inflammatory cytokines and chemokines by HDPF when challenged with bacterial antigens normally present in advanced caries lesions.
Triplicate HDPF from 4 children (n = 4; 2 boys and 2 girls) were cultured by explant technique and challenged or not with Escherichia coli lipopolysaccharide/1 μg/mL (EcLPS) or Enterococcus faecalis lipoteichoic acid/1 μg/mL (EfLTA) for 6 and 24 h. Most of published studies employed immortalized cells, i.e., without checking possible gender and genetic variables. mRNA expression and protein production were evaluated by RT-qPCR and ELISA MILLIPLEX®, respectively, for Interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17, Chemokine C-C motif ligand 2/monocyte chemoattractant protein 1 (CCL2/MCP-1), Chemokine C-C motif ligand 3/macrophage inflammatory protein 1-alpha (CCL3/MIP1-α), Chemokine C-C motif ligand 5/ regulated on activation, normal T cell expressed and secreted (CCL5/RANTES), C-X-C motif chemokine 12/ stromal cell-derived factor 1 (CXCL12/SDF-1), Tumor Necrosis Factor-alpha (TNF-α), Interferon-gamma (IFN γ), Vascular Endothelial Growth Factor (VEGF), Colony stimulating factor 1 (CSF-1) and Macrophage colony-stimulating factor (M-CSF).
EcLPS increased IL-1α, IL-1β, IL-8, CCL2, CCL5, TNF-α and CSF-1 mRNA and protein levels while EfLTA was only able to positively regulate gene expression and protein production of IL-8.
The results of the present study confirmed our hypothesis, since pulp fibroblasts from deciduous teeth are capable of increasing gene expression and protein production after being stimulated with EcLPS and EfLTA.
人们熟知口腔成纤维细胞对细菌刺激的免疫反应。然而，关于乳牙髓细胞（HDPF）的研究较少，而这些细胞对于儿童牙髓感染的治疗非常重要。本研究旨在评估 HDPF 受到通常存在于进展性龋损中的细菌抗原刺激时，细胞因子和趋化因子的表达和产生情况。
通过组织块培养法从 4 名儿童（n=4；2 男 2 女）中分离出 3 组 HDPF，并分别用大肠杆菌脂多糖/1μg/mL（EcLPS）或粪肠球菌脂磷壁酸/1μg/mL（EfLTA）刺激 6 或 24h。大多数已发表的研究都使用了永生化细胞，即没有检查可能的性别和遗传变量。通过 RT-qPCR 和 ELISA MILLIPLEX®分别评估白细胞介素（IL）-1α、IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12、IL-17、趋化因子 C-C 基序配体 2/单核细胞趋化蛋白 1（CCL2/MCP-1）、趋化因子 C-C 基序配体 3/巨噬细胞炎性蛋白 1-α（CCL3/MIP1-α）、趋化因子 C-C 基序配体 5/活化正常 T 细胞表达和分泌因子（RANTES）、C-X-C 基序趋化因子 12/基质细胞衍生因子 1（CXCL12/SDF-1）、肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFNγ）、血管内皮生长因子（VEGF）、集落刺激因子 1（CSF-1）和巨噬细胞集落刺激因子（M-CSF）的 mRNA 和蛋白表达。
EcLPS 增加了 IL-1α、IL-1β、IL-8、CCL2、CCL5、TNF-α 和 CSF-1 的 mRNA 和蛋白水平，而 EfLTA 仅能正向调节 IL-8 的基因表达和蛋白产生。
本研究结果证实了我们的假设，即乳牙髓细胞在受到 EcLPS 和 EfLTA 刺激后，能够增加基因表达和蛋白产生。
