Is collagen breakdown during periodontitis linked to inflammatory cells and expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gingival tissue?
BACKGROUND
Evidence of the role of matrix metalloproteinases (MMPs) produced by resident and inflammatory cells in periodontal destruction is now well established. The purpose of this study was to quantify, in healthy and diseased upper gingival connective tissue, the area fraction (AA%) occupied by collagen fibers, the cell number belonging to inflammatory cell subsets, and the amounts of MMPs and TIMPs (tissue inhibitors of MMPs) in order to investigate the possible correlations, if any, between such molecules, collagen loss, and inflammatory cell subsets.


METHODS
Gingival tissue specimens from 6 healthy controls (C) and 6 patients with severe periodontitis (P) were divided into 2 groups. The first group of specimens was frozen and used for the staining of collagen fibers by sirius red F3Ba and for immunohistochemistry with antibodies against CD8, CD4, CD22, CD68, and TIA-1 molecules. The second group was used for organ culture, zymography, Western blotting, and dot blotting. Morphometric and automated image analysis was performed for the evaluation of the area fraction occupied by collagen fibers, the number of inflammatory cell subsets and for enzymatic activities developed by MMPs, and the amounts of TIMPs expressed during periodontal disease.


RESULTS
In group P, the area fraction of collagen fibers (33 +/- 10%) was significantly decreased (P < 0.0002) when compared to group C (60 +/- 7%), and was correlated with the number of all inflammatory cells and amounts of MMPs and TIMPs. In group P, there were significant increases of CD8+, CD22+, CD68+, and TIA-1+ cells, as well as increases in the amounts of MMP-1, MMP-2, MMP-3, MMP-9, and the active form of MMP-9. The active form of MMP-9 and the amount of TIMP-1 were positively correlated with the number of CD22+, CD68+, and TIA-1+ cells.


CONCLUSIONS
The present study showed an imbalance between MMPs and TIMPs associated with the pathologic breakdown of the extracellular matrix during periodontitis. The active form of MMP-9 could be a marker for the clinical severity of periodontal disease.
Séguier
S
S
Départment d'Anatomie Pathologique, Faculté de Chirurgie Dentaire-Paris V, Montrouge, France.
Gogly
B
B
Bodineau
A
A
Godeau
G
G
Brousse
N
N
D000293:Adolescent; D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000032:analysis; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D001391:Azo Compounds; D001402:B-Lymphocytes / Q000473:pathology; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000473:pathology; D018414:CD8-Positive T-Lymphocytes / Q000473:pathology; D015815:Cell Adhesion Molecules; D003094:Collagen / Q000378:metabolism*; D004396:Coloring Agents; D046508:Culture Techniques; D005260:Female; D005881:Gingiva / Q000201:enzymology*; D016179:Granulocyte Colony-Stimulating Factor / Q000032:analysis; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D037102:Lectins; D007963:Leukocytes, Mononuclear / Q000378:metabolism* / Q000473:pathology; D008264:Macrophages / Q000473:pathology; D008297:Male; D020782:Matrix Metalloproteinases / Q000032:analysis / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis; D008875:Middle Aged; D009000:Monocytes / Q000473:pathology; D010518:Periodontitis / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D010586:Phagocytes / Q000378:metabolism* / Q000473:pathology; D039102:Poly(A)-Binding Proteins; D011506:Proteins; D016601:RNA-Binding Proteins / Q000032:analysis; D051918:Sialic Acid Binding Ig-like Lectin 2; D013223:Statistics as Topic; D000074019:T-Cell Intracellular Antigen-1; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis / Q000378:metabolism*; D000097382:CD68 Molecule
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D015214:Antigens, Differentiation, Myelomonocytic; D001391:Azo Compounds; D015815:Cell Adhesion Molecules; D003094:Collagen; D004396:Coloring Agents; D016179:Granulocyte Colony-Stimulating Factor; D037102:Lectins; D020782:Matrix Metalloproteinases; D008565:Membrane Proteins; D039102:Poly(A)-Binding Proteins; D011506:Proteins; D016601:RNA-Binding Proteins; D051918:Sialic Acid Binding Ig-like Lectin 2; D000074019:T-Cell Intracellular Antigen-1; D019714:Tissue Inhibitor of Metalloproteinases; C497867:CD22 protein, human; C067980:CD68 antigen, human; D000097382:CD68 Molecule; C071558:TIA1 protein, human; C009798:C.I. direct red 80