Anti‐adhesive and pro‐apoptotic effects of 2‐hydroxyethyl methacrylate on human gingival fibroblasts co‐cultured with Streptococcus mitis strains

Abstract

AimTo evaluate and observe the cellular reactions that occur during the interaction/integration between 2-hydroxyethyl methacrylate/host tissue/microbial environment, in a co-culture of human gingival fibroblasts (HGF) and Streptococcus mitis strains.MethodologyStreptococcus mitis were cultured with strains in the presence of 3 mmol L−1 HEMA for 48 h and 72 h. Cytotoxicity was evaluated by the trypan blue dye exclusion test. Apoptosis was evaluated by TUNEL analysis. Adhesion was evaluated by immunofluorescence and western blot analyses. Quantitative analyses of the results were acquired by Qwin Plus 3.5 and QuantityOne I-D analysis software, respectively. The statistical significance of the results was evaluated using t-tests and linear regression tests.ResultsThe trypan blue dye test revealed 47.3% and 46.5% of dead fibroblasts after 48 and 72 h HEMA treatment, respectively, while bacterial viability was not influenced by the presence of HEMA and fibroblasts. The expression of pro-collagen I, involved in fibroblast adhesion, in untreated samples ranged from 12.49% to 6.91% of the positive area after 48 and 72 h, respectively, dropping to below 2% of the positive area in the other experimental conditions. Unlike the trypan blue test, co-cultured samples treated with HEMA showed 20% and 25% versus 17% and 21% (after 48 and 72 h, respectively) of apoptotic cells.ConclusionsThe evidence for HEMA toxicity and anti-adhesive effects against eukaryotic cells was reduced in the presence of bacteria, suggesting that dental resins should be well polymerized to avoid the spread of toxic monomers within the mouth.

pPKC α regulates through integrin β 1 human gingival fibroblasts/Streptococcus mitis adhesion in response to HEMA

“…), little is known about the effect exerted by materials, such as HEMA released from dental composite in a coculture model which mimics in vitro the situation occurring in vivo (Zara et al . , Di Giulio et al . ).…”

Involvement of mitochondrial signalling pathway in HGFs/S. mitiscoculture response to TEGDMA treatment

“…Moreover, TEGDMA can severely disturb the odontogenic differentiation potential of pulp/progenitor cell, which might have significant consequences for pulp tissue homeostasis and repair . In this study, TEGDMA was administered for 48 h at a concentration of 1 m M to investigate the effects of chronic treatment on HGFs in a coculture system which mimics in vitro the situation occurring in vivo . In this experimental model, the choice of the S. mitis strain is of particular interest not only because it is a physiological commensals of the human oropharynx and can use a variety of strategies for the colonization of the oral cavity, but especially because it also expresses many virulent factors, inducing pathogenesis .…”

Biological Responses of Human Gingival Fibroblasts (HGFs) in an Innovative Co-Culture Model with Streptococcus mitis to Thermosets Coated with a Silver Polysaccharide Antimicrobial System

“…Thus, the aim of this study was to investigate cytotoxicity, migration, morphology and inflammatory cytokine production of HGFs grown on BisGMA-TEGDMA thermosets coated with Chitlac-nAg and co-cultured with S. mitis in the presence of saliva [23], in order to clarify the biological reactions occurring between biomaterials, host tissue and microbial environment.…”