title: Zimography is an effective method for detection of matrix metalloproteinase 2 (MMP-2) activity in cultured human fibroblasts.
abstract: OBJECTIVE
  To describe a method to characterize the gelatinase activity of cultured human periodontal fibroblasts stimulated with Pam3Cys and E. coli LPS, ligands of TLR2 and TLR4 respectively, and by centrifugation of the cultures, simulating an orthodontic force.
  
  
  METHODS
  To study MMP-2 activity, primary cultures of human periodontal fibroblasts were stimulated with the addition of TLRs 2 and 4 ligands and the application of mechanical force by centrifugation at 141 x g for 30 min. Supernatant media was collected 24 hours later to perform protein quantification and zymography.
  
  
  RESULTS
  MMP-2 activity suffered an increase in cultures co-stimulated with TLRs 2 and 4 ligands alone or with the presence of mechanical force application compared to basal levels.
  
  
  CONCLUSIONS
  Zymography, one of the several methods to study MMPs activities, is a simple, qualitative and efficient method based on electrophoresis of bis-acrylamide gels copolymerized with a protein substrate.
authors.affiliation: Department of Pathology, School of Medicine, UFMG, Belo Horizonte-MG, Brazil.
authors.forename: Rodolfo Assis
authors.initials: RA
authors.lastname: Lisboa
authors.forename: Marcus Vinícius
authors.initials: MV
authors.lastname: Andrade
authors.forename: José Renan
authors.initials: JR
authors.lastname: Cunha-Melo
mesh_terms: D002470:Cell Survival; D002478:Cells, Cultured; D004586:Electrophoresis / Q000379:methods*; D005347:Fibroblasts / Q000201:enzymology*; D018093:Gelatinases / Q000502:physiology; D006801:Humans; D008074:Lipoproteins; D020778:Matrix Metalloproteinase 2 / Q000032:analysis* / Q000502:physiology; D010513:Periodontal Ligament / Q000166:cytology; D015203:Reproducibility of Results; D018709:Statistics, Nonparametric; D013997:Time Factors; D051193:Toll-Like Receptors / Q000032:analysis
chemical_list: D008074:Lipoproteins; D051193:Toll-Like Receptors; C061108:N-palmitoyl-S-(2,3-bis(palmitoyloxy)propyl)cysteinyl-seryl-lysyl-lysyl-lysyl-lysine; D018093:Gelatinases; D020778:Matrix Metalloproteinase 2