Response of periodontal ligament stem cells to lipopolysaccharide and calcium silicate-based materials

Abstract

This study was conducted to assess the in vitro response of human periodontal ligament stem cells (hPDLSCs) to bacterial lipopolysaccharide (LPS) activation and application of three calcium silicate-based materials (CSBM): Bio-C Sealer, MTA Fillapex and Cimmo HP. Characterization of the CSBM was performed by FTIR (n = 3). Extracts of Bio-C Sealer, MTA Fillapex and Cimmo HP were prepared and diluted (1:1, 1:4 and 1:16). Culture of hPDLSCs was established and treated or not with LPS from Escherichia coli (1 µg/mL) for 7 days. MTT assay was used to assess cell viability at 24, 48 and 72 h (n = 9). Alkaline phosphatase (ALP) activity was indirectly assayed at day 7 (n = 5). TNF-α and Il -1 0 cytokines were quantified by ELISA at 24h-cell supernatants (n = 6). Data were analyzed by ANOVA and Tukey’s test (α = 0.05). The cell viability of the LPS-activated hPDLSCs were higher than untreated control (p < 0.05). The application of CSBM affected the cell viability of untreated and LPS-activated cells (p < 0.05). ALP activity was higher for Bio-C Sealer and Cimmo HP in untreated and LPS-activated cells, respectively (p < 0.05). Application of CSBM normalized the TNF-α secretion in the LPS-activated cells (p < 0.05). Only MTA Fillapex in untreated hPDLSCs presented higher values of Il -1 0 (p < 0.05). Taken collectively, the results suggests that the simulation of the inflammatory process by LPS affect the in vitro response the hPDLSCs to the application of the CSBM.

Exposure to lipopolysaccharide and calcium silicate-based materials affects the behavior of dental pulp cells

“…In this study, cell viability was assessed through an MTT assay. Even though this method measures metabolic activity and does not differentiate cell cycle, anti-proliferative effects, cell apoptosis, or necrosis, It is a reliable and accessible method to assess the cell viability of endodontic materials (4). We also worked with the CSBM extracts to infer a possible dose dilution that occurs in vivo (4) as well as the effects of these materials at a distance (11).…”

“…The absorbance at 590 nm was measured (Synergy HT, Biotek, Instruments, Inc. Winooski, VT, USA). ALP activity was normalized by the total protein content and expressed as μmol of thymolphthalein/h/mg of protein (4).…”