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Chronic exposure to lipopolysaccharides as an in vitro model to simulate the impaired odontogenic potential of dental pulp cells under pulpitis conditions

Soares, Igor Paulino Mendes; Anselmi, Caroline; Pires, Maria Luiza Barucci Araujo; Ribeiro, Rafael Antonio De Oliveira; Leite, Maria Lu��Sa; Soares, Diana Gabriela; Costa, Carlos Alberto De Souza; Hebling, Josimeri

Journal of Applied Oral Science; v. 31 (2023)

Universidade de S��o Paulo. Faculdade de Odontologia de Bauru 2023-08-14

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  • Título:
    Chronic exposure to lipopolysaccharides as an in vitro model to simulate the impaired odontogenic potential of dental pulp cells under pulpitis conditions
  • Autor: Soares, Igor Paulino Mendes; Anselmi, Caroline; Pires, Maria Luiza Barucci Araujo; Ribeiro, Rafael Antonio De Oliveira; Leite, Maria Lu��Sa; Soares, Diana Gabriela; Costa, Carlos Alberto De Souza; Hebling, Josimeri
  • Assuntos: Lipopolysaccharides; Cell Culture Techniques; Dental Pulp; Pulpitis; Biomineralization
  • É parte de: Journal of Applied Oral Science; v. 31 (2023)
  • Descrição: Simulating a bacterial-induced pulpitis environment in vitro may contribute to exploring mechanisms and bioactive molecules to counteract these adverse effects. Objective: To investigate the chronic exposure of human dental pulp cells (HDPCs) to lipopolysaccharides (LPS) aiming to establish a cell culture protocol to simulate the impaired odontogenic potential under pulpitis conditions. Methodology: HDPCs were isolated from four healthy molars of different donors and seeded in culture plates in a growth medium. After 24 h, the medium was changed to an odontogenic differentiation medium (DM) supplemented or not with E. coli LPS (0 - control, 0.1, 1, or 10 ��g/mL) (n=8). The medium was renewed every two days for up to seven days, then replaced with LPS-free DM for up to 21 days. The activation of NF-��B and F-actin expression were assessed (immunofluorescence) after one and seven days. On day 7, cells were evaluated for both the gene expression (RT-qPCR) of odontogenic markers (COL1A1, ALPL, DSPP, and DMP1) and cytokines (TNF, IL1B, IL8, and IL6) and the production of reactive nitrogen (Griess) and oxygen species (Carboxy-H2DCFDA). Cell viability (alamarBlue) was evaluated weekly, and mineralization was assessed (Alizarin Red) at 14 and 21 days. Data were analyzed with ANOVA and post-hoc tests (��=5%). Results: After one and seven days of exposure to LPS, NF-��B was activated in a dose-dependent fashion. LPS at 1 and 10 ��g/mL concentrations down-regulated the gene expression of odontogenic markers and up-regulated cytokines. LPS at 10 ��g/mL increased both the production of reactive nitrogen and oxygen species. LPS decreased cell viability seven days after the end of exposure. LPS at 1 and 10 ��g/mL decreased hDPCs mineralization in a dose-dependent fashion. Conclusion: The exposure to 10 ��g/mL LPS for seven days creates an inflammatory environment that is able to impair by more than half the odontogenic potential of HDPCs in vitro, simulating a pulpitis-like condition.
  • DOI: 10.1590/1678-7757-2023-0032
  • Títulos relacionados: https://www.revistas.usp.br/jaos/article/view/215010/197216
  • Editor: Universidade de S��o Paulo. Faculdade de Odontologia de Bauru
  • Data de criação/publicação: 2023-08-14
  • Formato: Adobe PDF
  • Idioma: Inglês
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