Expression of matrix metalloproteinases in healthy and diseased human gingiva.
OBJECTIVE
The aim of our study was to investigate the patterns of several metalloproteinases (MMP-1, MMP-2 and MT1-MMP) mRNAs expression using a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and to correlate them with clinical parameters and bacteriological diagnosis in healthy versus diseased human gingiva.


METHODS
To identify the cell origin of MMP production, in situ hybridization (ISH) was also performed for the MMPs on the same samples. 17 gingival biopsies were collected (13 affected by advanced periodontitis and 4 healthy used as controls) and plaque index, gingival index, pocket depth and bleeding on probing were measured. Subgingival microbial samples were also collected to be analysed by a DNA probe technique. The biopsies were processed both for RT-PCR and ISH. We also investigated a model for bacterial induced MMP expression in human gingival fibroblasts (HGF) infected by Eikenella corrodens.


RESULTS
We found an expression of the mRNA encoding MMP-1 only in diseased gingiva but at low levels relative to beta-actin (mean+/-SD: diseased versus healthy: 0.013+/-0.024 versus 0). Although the frequencies and levels of mRNA encoding for MMP-2 or MT1-MMP are not significantly different between each group (mean+/-SD: 0.329+/-0.344 versus 0.137+/-0.219 for MMP-2; 0.485+/-0.374 versus 0.466+/-0.296 for MT1-MMP), using ISH, we observed an expression of both mRNAs in fibroblasts of pathological specimens at sites that histologically showed signs of chronic inflammation and connective tissue remodelling. In vitro infection of HGF by Eikenella corrodens stimulated 3-fold the production of the mRNA encoding MMP-2 while other mRNAs remained unchanged.


CONCLUSIONS
Our results did not reveal significant differences in the expression of mRNAs encoding for the MMPs between healthy and periodontitis-affected patients, reflecting the great heterogeneity in the periodontal status of individuals. However, they indicate that gingival fibroblasts are an active source of MMP-2 production in response to a periopathogen.
Dahan
M
M
INSERM U 424, Fédération de Recherches Odontologiques, Strasbourg, France.
Nawrocki
B
B
Elkaïm
R
R
Soell
M
M
Bolcato-Bellemin
A L
AL
Birembaut
P
P
Tenenbaum
H
H
D000293:Adolescent; D000328:Adult; D000704:Analysis of Variance; D001421:Bacteria, Anaerobic / Q000302:isolation & purification / Q000472:pathogenicity; D016022:Case-Control Studies; D015169:Colony Count, Microbial; D016124:Eikenella corrodens / Q000472:pathogenicity; D005260:Female; D005347:Fibroblasts / Q000201:enzymology / Q000382:microbiology; D005881:Gingiva / Q000201:enzymology*; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis; D020782:Matrix Metalloproteinases / Q000096:biosynthesis*; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000096:biosynthesis; D008875:Middle Aged; D010518:Periodontitis / Q000201:enzymology* / Q000382:microbiology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric
D012333:RNA, Messenger; D020782:Matrix Metalloproteinases; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1