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Human Neutrophil Gelatinase and Associated Lipocalin in Adult and Localized Juvenile Periodontitis

Department of Periodontology, Institute of Dentistry, PO Box 41, FIN-00014 University of Helsinki, Helsinki, Finland

T. Ingman

Department of Periodontology, Institute of Dentistry, PO Box 41, FIN-00014 University of Helsinki, Helsinki, Finland, Department of Pedodontics and Orthodontics, Institute of Dentistry, University of Helsinki, Finland

P.-L. Lukinmaa

Department of Oral Pathology, Institute of Dentistry, University of Helsinki, Helsinki, Finland

T. Salo

Department of Oral Surgery, Institute of Dentistry, University of Oulu, Oulu, Finland

L. Kjeldsen

Granulocyte Research Laboratory, Rigshospitalet, Copenhagen, Denmark

N. Borregaard

Granulocyte Research Laboratory, Rigshospitalet, Copenhagen, Denmark

L. Tjäderhane

Department of Cariology, Institute of Dentistry, University of Oulu, Oulu, Finland

Y.T. Konttinen

Department of Anatomy, University of Helsinki, Helsinki, Finland, Fourth Department of Medicine, University of Helsinki, Helsinki, Finland

T. Sorsa

Department of Periodontology, Institute of Dentistry, PO Box 41, FIN-00014 University of Helsinki, Helsinki, Finland

In search of direct in vivo evidence of matrix metalloproteinases (MMPs) in periodontal tissue destruction, we studied the presence and localization of MMP-9 and neutrophil gelatinase-associated lipocalin (NGAL) in adult periodontitis (AP) and localized juvenile periodontitis (LJP) gingival tissue specimens by immunohistochemistry, and the activities of gelatinases by Western blot, enzymography, and activity measurements, using radioactive gelatin as substrate in gingival crevicular fluid (GCF) and saliva. In gingival tissue obtained from AP and LJP patients, polymorphonuclear leukocyte (PMN) 92-kDa MMP-9 and NGAL were seen in the connective tissue, but both the sulcular and the oral epithelia were consistently negative. Whereas PMNs located in the gingival blood vessels showed strictly cytoplasmic MMP-9 and NGAL immunoreactivities, in the case of PMN extravasation the staining reactions extended extracellularly. Gelatinase activities consisting mainly of 92-kDa gelatinase were increased in AP GCF relative to LJP GCF and periodontally healthy control GCF. Western blot with specific anti-NGAL antibodies revealed the presence of 25-kDa NGAL and its high-molecular-weight forms in AP and LJP GCF and saliva and in culture medium of oral keratinocytes, but not in gingival fibroblast culture medium. We conclude that extravasated degranulating PMNs are the major source of MMP-9 and NGAL in periodontitis gingiva, GCF, and saliva.

Key Words: gingiva • periodontitis • matrix metalloproteinases • neutrophil gelatinase-associated lipocalin • immunohistochemistry • Received April 12 • 1995 • Accepted April 4 • 1996

Journal of Dental Research, Vol. 75, No. 8, 1553-1563 (1996)
DOI: 10.1177/00220345960750080601


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