Gingival crevicular fluid can degrade Emdogain and inhibit Emdogain-induced proliferation of periodontal ligament fibroblasts.
OBJECTIVE
Emdogain (EMD), consisting mostly of amelogenin, is used in periodontal therapy to regenerate lost connective tissue. Emdogain is applied onto periodontally affected root surfaces, where it becomes exposed to proteolytic enzymes. In this study, we aimed to find out whether gingival crevicular fluid or matrix metalloproteinases (MMPs) could degrade EMD, and whether this degradation has consequences for in vitro cell proliferation.


METHODS
We studied the effects of 156 gingival crevicular fluid samples collected from subjects with different stages of periodontal disease and from healthy control subjects and the effects of MMP-1, -2, -8, -9, -13 and -14 on the degradation of EMD using EMD-embedded zymography. The effects of gingival crevicular fluid with or without EMD and the effects of amelogenin on the proliferation of cultured periodontal ligament fibroblasts were studied by cell proliferation enzyme-linked immunosorbent assay kit.


RESULTS
Degradation of Emdogain induced by gingival crevicular fluid was greater in samples from all stages of periodontal diseases compared with healthy control samples. Of the MMPs studied, only MMP-2 and MMP-8 showed limited EMD-degrading activities. One hundred micrograms per millilitre of EMD increased proliferation of periodontal ligament fibroblasts on average by 24% (confidence interval 0.60-0.64) and at 200 microg/mL by 30% (confidence interval 0.62-0.68) compared with control fibroblasts (confidence interval 0.48-0.52). However, gingival crevicular fluid (10 microg/mL) together with 100 microg/mL EMD induced the proliferation only by 6% (confidence interval 0.51-0.55) and with 200 microg/mL EMD by 12% (confidence interval 0.54-0.58). Amelogenin at 200 microg/mL decreased the proliferation of periodontal ligament fibroblasts by 54% (confidence interval 0.22-0.25).


CONCLUSIONS
We suggest that diseased gingival crevicular fluid containing various proteases leads to degradation of EMD and decreased proliferation of periodontal ligament fibroblasts.
Laaksonen
M
M
Department of Oral and Maxillofacial Diseases, Institute of Dentistry, Helsinki University Central Hospital, University of Helsinki, Helsinki, Finland.
Salo
T
T
Vardar-Sengul
S
S
Atilla
G
G
Saygan
B Han
BH
Simmer
J P
JP
Baylas
H
H
Sorsa
T
T
D000293:Adolescent; D000328:Adult; D010520:Aggressive Periodontitis / Q000378:metabolism; D016301:Alveolar Bone Loss / Q000378:metabolism; D053523:Amelogenin / Q000378:metabolism / Q000494:pharmacology; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D055113:Chronic Periodontitis / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000378:metabolism* / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D005883:Gingival Crevicular Fluid / Q000201:enzymology / Q000378:metabolism*; D005884:Gingival Hemorrhage / Q000378:metabolism; D005891:Gingivitis / Q000378:metabolism; D006801:Humans; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000494:pharmacology; D053509:Matrix Metalloproteinase 13 / Q000494:pharmacology; D053511:Matrix Metalloproteinase 14 / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000494:pharmacology; D020784:Matrix Metalloproteinase 8 / Q000494:pharmacology; D020780:Matrix Metalloproteinase 9 / Q000494:pharmacology; D008875:Middle Aged; D017622:Periodontal Attachment Loss / Q000378:metabolism; D010510:Periodontal Diseases / Q000378:metabolism; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects*; D010514:Periodontal Pocket / Q000378:metabolism; D055815:Young Adult
D053523:Amelogenin; D003746:Dental Enamel Proteins; C109069:enamel matrix proteins; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; C488762:MMP8 protein, human; D020784:Matrix Metalloproteinase 8; D020780:Matrix Metalloproteinase 9; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14