Zimography is an effective method for detection of matrix metalloproteinase 2 (MMP-2) activity in cultured human fibroblasts
Acta cir. bras
;
28(3): 216-220, Mar. 2013. ilus
Article
in English
| LILACS
| ID: lil-667933
ABSTRACT
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.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Electrophoresis
/
Fibroblasts
Type of study:
Diagnostic study
/
Qualitative research
Limits:
Humans
Language:
English
Journal:
Acta cir. bras
Journal subject:
General Surgery
/
Procedimentos Cir£rgicos Operat¢rios
Year:
2013
Type:
Article
Affiliation country:
Brazil
Institution/Affiliation country:
UFMG/BR
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