Matrix metalloproteinase-8 activity in gingival crevicular fluid: development of a novel assay.
OBJECTIVE
The matrix metalloproteinases (MMPs) play a role in regulating turnover and metabolism of connective tissues in health but they have also been implicated in a wide variety of pathological conditions, including periodontal disease. MMP-8 has been extensively studied in periodontal health and disease using ELISA, although this technique is limited by its inability to determine enzyme activity. The aim was to develop an assay specifically to measure MMP-8 activity and to demonstrate its use in the analysis of gingival crevicular fluid samples.


METHODS
A specific antibody was used to coat black 96-well microtitre plates to capture MMP-8 selectively. The activity of bound MMP-8 was measured using a fluorogenic substrate. Gingival crevicular fluid samples, from healthy and periodontally diseased sites, were collected using PerioPaper strips and tested for MMP-8 activity.


RESULTS
Significantly higher MMP-8 activity was demonstrated in gingival crevicular fluid from periodontally diseased sites compared with healthy sites that exhibited basal or no MMP-8 activity. No cross-reactivity with other MMPs was noted.


CONCLUSIONS
We show, for the first time, that MMP-8 activity can be specifically detected and quantified in gingival crevicular fluid samples. Measurement of MMP-8 activity could prove to be useful in monitoring periodontal disease progression.
Centre for Experimental Medicine, Queen's University Belfast, Belfast, UK.
M T C
MTC
Mc Crudden
Centre for Dental Education, Queen's University Belfast, Belfast, UK.
C R
CR
Irwin
Centre for Experimental Medicine, Queen's University Belfast, Belfast, UK.
I
I
El Karim
Centre for Public Health, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK.
G J
GJ
Linden
Centre for Experimental Medicine, Queen's University Belfast, Belfast, UK.
F T
FT
Lundy
D000328:Adult; D000368:Aged; D000906:Antibodies / Q000276:immunology; D055113:Chronic Periodontitis / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005883:Gingival Crevicular Fluid / Q000201:enzymology*; D006801:Humans; D020784:Matrix Metalloproteinase 8 / Q000276:immunology / Q000378:metabolism*; D008875:Middle Aged
D000906:Antibodies; D020784:Matrix Metalloproteinase 8
gingival crevicular fluid; matrix metalloproteinase; periodontal disease; periodontitis