Matrix metalloproteinase‐8 activity in gingival crevicular fluid: development of a novel assay

• Published in: Journal of periodontal research Vol. 52; no. 3; pp. 556 - 561
• Main Authors: Mc Crudden, M. T. C., Irwin, C. R., El karim, I., Linden, G. J., Lundy, F. T.
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01-06-2017
• Subjects: Adult; Aged; Antibodies - immunology; Chronic Periodontitis - enzymology; Connective tissues; Cross-reactivity; Dentistry; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Gingiva; gingival crevicular fluid; Gingival Crevicular Fluid - enzymology; Humans; Matrix metalloproteinase; Matrix Metalloproteinase 8 - immunology; Matrix Metalloproteinase 8 - metabolism; Metalloproteinase; Middle Aged; Neutrophil collagenase; periodontal disease; Periodontics; periodontitis; gingival crevicular fluid; matrix metalloproteinase; periodontal disease; periodontitis
• ISSN: 0022-3484; 1600-0765
• DOI: 10.1111/jre.12423

Background and 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.
Material and 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.
Conclusion
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.