Osteogenic potential of periodontal ligament stem cells are unaffected after exposure to lipopolysaccharides.
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.
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Mayra Laino
ML
Albiero
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Bruna Rabelo
BR
Amorim
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Márcio Zaffalon
MZ
Casati
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Enilson Antonio
EA
Sallum
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Francisco Humberto
FH
Nociti
Universidadade de Campinas - UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontics, Piracicaba, São Paulo, Brazil.
Karina Gonzales
KG
Silvério
D000469:Alkaline Phosphatase / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D053583:Interleukin-1beta / Q000032:analysis; D008070:Lipopolysaccharides / Q000633:toxicity*; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000032:analysis; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000187:drug effects*; D010513:Periodontal Ligament / Q000166:cytology*; D016966:Porphyromonas gingivalis; D060888:Real-Time Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D013997:Time Factors; D051193:Toll-Like Receptors / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D050659:Core Binding Factor Alpha 1 Subunit; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D050814:Octamer Transcription Factor-3; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; D015675:Osteocalcin; D000469:Alkaline Phosphatase