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Zimography is an effective method for detection of matrix metalloproteinase 2 (MMP-2) activity in cultured human fibroblasts.
Lisboa, Rodolfo Assis; Andrade, Marcus Vinícius; Cunha-Melo, José Renan.
Afiliação
  • Lisboa RA; Department of Pathology, School of Medicine, UFMG, Belo Horizonte-MG, Brazil.
Acta Cir Bras ; 28(3): 216-20, 2013 Mar.
Article em En | MEDLINE | ID: mdl-23503864
PURPOSE: 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. CONCLUSION: 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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Metaloproteinase 2 da Matriz / Eletroforese / Fibroblastos Tipo de estudo: Diagnostic_studies / Qualitative_research Limite: Humans Idioma: En Revista: Acta Cir Bras Ano de publicação: 2013 Tipo de documento: Article País de afiliação: Brasil País de publicação: Brasil

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Metaloproteinase 2 da Matriz / Eletroforese / Fibroblastos Tipo de estudo: Diagnostic_studies / Qualitative_research Limite: Humans Idioma: En Revista: Acta Cir Bras Ano de publicação: 2013 Tipo de documento: Article País de afiliação: Brasil País de publicação: Brasil