Characteristics of collagenase-2 from gingival crevicular fluid and peri-implant sulcular fluid in periodontitis and peri-implantitis patients: pilot study.
OBJECTIVE
To compare collagenase activity and collagenolytic matrix metalloproteinase (MMP) levels in gingival crevicular fluid (GCF) and in peri-implant sulcular fluid (PISF) in gingivitis (G), chronic periodontitis (CP), and peri-implantitis (PI) human subjects.


METHODS
GCF and PISF were collected on filter paper strips, volume was determined, and samples were extracted in buffer containing general proteinase but not MMP inhibitors. Collagenase activity was measured using a DNP-synthetic octapeptide, and molecular and activation forms of collagenase-2 by Western immunoblotting.


RESULTS
GCF from CP and G sites exhibited elevated collagenase activity and flow, but collagenase concentrations expressed per microl were not significantly different between the healthy and G sites. Minimal fluid was obtained from healthy PISF, and collagenase concentration was the same or lower than in healthy GCF. Although PISF flow was 34% lower than GCF flow in CP subjects, collagenase concentration in CP and in PI sites was 78% and 971% greater, respectively, than in the appropriate healthy sites. Western immunoblot revealed MMP-8 in both PISF and GCF; fibroblast-type MMP-8 was not detected in healthy GCF and PISF. Immunoreactivity level and inactive and activated forms of PMN-type MMP-8 in GCF and PISF increased with the severity of periodontitis and peri-implantitis. Enhanced levels of fibroblast-type MMP-8 in active form were detected only in severe CP GCF and PI PISF.


CONCLUSIONS
Peri-implantitis PISF contained higher collagenase-2 levels and activity than GCF from similar deep CP sites. GCF and PISF from severe CP and PI exhibited the highest activation of MMP-8 isoenzymes species (PMN and fibroblast-type).
Xu
Ling
L
Department of General Dentistry, School of Dental Medicine, Stony Brook University, New York, Stony Brook, USA.
Yu
Zhao
Z
Lee
Hsi-Ming
HM
Wolff
Mark S
MS
Golub
Lorne M
LM
Sorsa
Timo
T
Kuula
Heidi
H
D000328:Adult; D000368:Aged; D016301:Alveolar Bone Loss / Q000201:enzymology; D015153:Blotting, Western; D002908:Chronic Disease; D015921:Dental Implants; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005881:Gingiva / Q000201:enzymology; D005883:Gingival Crevicular Fluid / Q000201:enzymology*; D005884:Gingival Hemorrhage / Q000201:enzymology; D005891:Gingivitis / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000032:analysis; D008297:Male; D020784:Matrix Metalloproteinase 8 / Q000032:analysis*; D008875:Middle Aged; D009504:Neutrophils / Q000201:enzymology; D017622:Periodontal Attachment Loss / Q000201:enzymology; D010514:Periodontal Pocket / Q000201:enzymology; D010518:Periodontitis / Q000201:enzymology*; D010519:Periodontium / Q000201:enzymology; D010865:Pilot Projects
D015921:Dental Implants; D007527:Isoenzymes; C488762:MMP8 protein, human; D020784:Matrix Metalloproteinase 8