title: Anti-adhesive and pro-apoptotic effects of 2-hydroxyethyl methacrylate on human gingival fibroblasts co-cultured with Streptococcus mitis strains.
abstract: OBJECTIVE
  To 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.
  
  
  METHODS
  Streptococcus 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The 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.
authors: Department of Drug Sciences, University G. d'Annunzio Chieti-Pescara, Chieti, Italy. s.zara@unich.it
authors: S
authors: S
authors: Zara
authors: M
authors: M
authors: Di Giulio
authors: S
authors: S
authors: D'Ercole
authors: L
authors: L
authors: Cellini
authors: A
authors: A
authors: Cataldi
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001422:Bacterial Adhesion / Q000187:drug effects; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D016923:Cell Death / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D024042:Collagen Type I / Q000032:analysis / Q000187:drug effects; D004396:Coloring Agents; D003764:Dental Materials / Q000494:pharmacology* / Q000633:toxicity; D005347:Fibroblasts / Q000187:drug effects*; D005455:Fluorescent Antibody Technique; D005881:Gingiva / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D020287:In Situ Nick-End Labeling; D008689:Methacrylates / Q000494:pharmacology* / Q000633:toxicity; D050296:Microbial Viability / Q000187:drug effects; D034361:Streptococcus mitis / Q000187:drug effects*; D013997:Time Factors; D014343:Trypan Blue
chemical_list: D024042:Collagen Type I; D004396:Coloring Agents; D003764:Dental Materials; D008689:Methacrylates; C005044:hydroxyethyl methacrylate; D014343:Trypan Blue