Albiero Mayra Laino, Amorim Bruna Rabelo, Casati Márcio Zaffalon, Sallum Enilson Antonio, Nociti Francisco Humberto, Silvério Karina Gonzales
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Braz Oral Res. 2017 Jan 26;31:e17. doi: 10.1590/1807-3107BOR-2017.vol31.0017.
Periodontitis develops as a result of a continuous interaction between host cells and subgingival pathogenic bacteria. The periodontium has a limited capacity for regeneration, probably due to changes in periodontal ligament stem cells (PDLSCs) phenotype. The aim of this study was to evaluate the effects of lipopolysaccharides from Porphyromonas gingivalis (PgLPS) on mesenchymal phenotype and osteoblast/cementoblast (O/C) potential of PDLSCs. PDLSCs were assessed for Toll-like receptor 2 (TLR2) expression by immunostaining technique. After, cells were exposed to PgLPS, and the following assays were carried out: (i) cell metabolic activity using MTS; (ii) gene expression for IL-1β, TNF-α and OCT-4 by real-time polymerase chain reaction (RT-qPCR); (iii) flow cytometry for STRO-1 and CD105, and (iv) osteogenic differentiation. PDLSCs were positive for TLR2. PgLPS promoted cell proliferation, produced IL-1β and TNF-α, and did not affect the expression of stem cell markers, STRO-1, CD105 and OCT-4. Under osteogenic condition, PDLSCs exposed to PgLPS showed a similar potential to differentiate toward osteoblast/cementoblast phenotype compared to control group as revealed by mineralized matrix deposition and levels of transcripts for RUNX2, ALP and OCN. These results provide evidence that PgLPS induces pro-inflammatory cytokines, but does not change the mesenchymal phenotype and osteoblast/cementoblast differentiation potential of PDLSCs.
牙周炎是宿主细胞与龈下致病细菌持续相互作用的结果。牙周组织的再生能力有限，这可能是由于牙周膜干细胞（PDLSCs）表型的改变所致。本研究的目的是评估牙龈卟啉单胞菌脂多糖（PgLPS）对PDLSCs间充质表型和成骨细胞/成牙骨质细胞（O/C）潜能的影响。通过免疫染色技术评估PDLSCs中Toll样受体2（TLR2）的表达。之后，将细胞暴露于PgLPS，并进行以下检测：（i）使用MTS检测细胞代谢活性；（ii）通过实时聚合酶链反应（RT-qPCR）检测IL-1β、TNF-α和OCT-4的基因表达；（iii）对STRO-1和CD105进行流式细胞术检测；（iv）进行成骨分化检测。PDLSCs的TLR2呈阳性。PgLPS促进细胞增殖，产生IL-1β和TNF-α，且不影响干细胞标志物STRO-1、CD105和OCT-4的表达。在成骨条件下，与对照组相比，暴露于PgLPS的PDLSCs在矿化基质沉积以及RUNX2、ALP和OCN转录水平方面显示出类似的向成骨细胞/成牙骨质细胞表型分化的潜能。这些结果表明，PgLPS可诱导促炎细胞因子，但不会改变PDLSCs的间充质表型和成骨细胞/成牙骨质细胞的分化潜能。
