title: Evaluation of cytotoxicity and up-regulation of gelatinases in human fibroblast cells by four root canal sealers.
abstract: OBJECTIVE
  To investigate the effects of root canal sealers on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in human fibroblasts.
  
  
  METHODS
  Human fibroblasts (MRC5, 3×10(5) cells per well) were incubated directly or indirectly with AH Plus, Endomethasone N, Pulp Canal Sealer EWT or Sealapex for 30 min, 1, 4 or 24 h (time-points). The cytotoxicity of all root canal sealers was determined by counting viable cells using the trypan blue exclusion assay. Supernatants of cell cultures incubated with root sealers directly or indirectly were collected after each time-point to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and the Tukey's tests.
  
  
  RESULTS
  Cells secreted MMP-2 after periods of 4 and 24 h; however, there were no significant differences between the sealers. Secretion of gelatinases was elevated by root canal sealers in direct contact with the cell monolayer when compared to indirect contact (P < 0.05). At the time-points tested, no gelatinolytic activity could be detected in the control group without the sealers. The cytotoxicity results revealed that all sealers were cytotoxic in both contact forms. Sealapex had the lowest cytotoxicity and AH Plus the most cytotoxicity.
  
  
  CONCLUSIONS
  All root canal sealers induced the expression of MMP-2 in MRC5 fibroblasts. AH Plus had the highest cytotoxicity amongst the tested sealers, but all were associated with cytotoxic effects.
authors.affiliation: Department of Restorative Dentistry, Endodontic Division, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil.
authors.forename: E J N L
authors.initials: EJ
authors.lastname: Silva
authors.forename: T
authors.initials: T
authors.lastname: Accorsi-Mendonça
authors.forename: J F A
authors.initials: JF
authors.lastname: Almeida
authors.forename: C C R
authors.initials: CC
authors.lastname: Ferraz
authors.forename: B P F A
authors.initials: BP
authors.lastname: Gomes
authors.forename: A A
authors.initials: AA
authors.lastname: Zaia
mesh_terms: D002126:Calcium Hydroxide / Q000737:chemistry; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D003907:Dexamethasone / Q000737:chemistry; D004338:Drug Combinations; D004853:Epoxy Resins / Q000737:chemistry; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D005557:Formaldehyde / Q000737:chemistry; D018093:Gelatinases / Q000032:analysis / Q000187:drug effects*; D006801:Humans; D006854:Hydrocortisone / Q000737:chemistry; D008422:Materials Testing; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000187:drug effects; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000187:drug effects; D012387:Root Canal Filling Materials / Q000737:chemistry / Q000633:toxicity*; D012459:Salicylates / Q000737:chemistry; D013499:Surface Properties; D013943:Thymol / Q000031:analogs & derivatives / Q000737:chemistry; D013997:Time Factors; D014343:Trypan Blue; D015854:Up-Regulation / Q000187:drug effects*; D015035:Zinc Oxide-Eugenol Cement / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D004338:Drug Combinations; D004853:Epoxy Resins; D012387:Root Canal Filling Materials; D012459:Salicylates; D015035:Zinc Oxide-Eugenol Cement; C011244:corticosteroid methanetriol mixture; C534916:epoxy resin-based root canal sealer; C055601:Sealapex; D005557:Formaldehyde; D013943:Thymol; D003907:Dexamethasone; D018093:Gelatinases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D014343:Trypan Blue; D002126:Calcium Hydroxide; D006854:Hydrocortisone