HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text Full Text (PDF) Services Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (12) Citing Articles via Google Scholar Google Scholar Articles by Uitto, V.-J. Articles by Saarialho-Kere, U. Search for Related Content PubMed PubMed Citation Articles by Uitto, V.-J. Articles by Saarialho-Kere, U.

Matrilysin (Matrix Metalloproteinase-7) Expression in Human Junctional Epithelium

¹ Department of Biological and Medical Sciences, Faculty of Dentistry, University of British Columbia, 2199 Wesbrook mall, Vancouver, BC, V6T 1Z3, Canada;
² Department of Oral and Maxillofacial Diseases, Surgical Hospital, HUCH, Helsinki, Finland;
³ Turku Centre for Biomaterials, University of Turku, FIN-20520, Turku, Finland;
⁴ National Public Health Institute, Anaerobe Reference Laboratory, FIN-00300 Finland; and
⁵ Department of Dermatology, Helsinki University Central Hospital, Meilahdentie 2, FIN-00250 Helsinki, Finland;

View larger version (99K): [in a new window]   Figure 1. Matrilysin (MMP-7) expression in epithelium of human periodontium. Histological sections were prepared from normal periodontal tissue obtained during therapeutic removal of tooth 24 (A and C) and from a palatal explant tissue cultured on a Millipore filter for 5 days (B and D). Positive reaction with anti-matrilysin antibody (brown, indicated by arrows) can be observed in junctional epithelial cells facing the tooth (A) and in epithelial cell rests of Malassez (C). Suprabasal cells of the explant epibolus at the filter side also reacted with the anti-matrilysin antibody (B; a higher magnification in D). No reaction was detected in connective tissue (CT) in either the gingival tissue or the explant. Neither pocket epithelium (PE) nor infiltrated connective tissue (ICT) of periodontal pocket tissue reacted with matrilysin antibody (E and F).

View larger version (48K): [in a new window]   Figure 2. Up-regulation of matrilysin expression in epithelial cells cultured in the presence of oral bacteria. Bacteria (a stock solution, OD600 = 0.5) were added at a 1:50 dilution to semiconfluent porcine periodontal ligament epithelial cell cultures. After 24-hour culture in the absence of serum, total RNA (20 µg/sample) isolated from the cells was analyzed by Northern hybridization, with specific cDNA probes for human matrilysin and glyceraldehyde-3-phosphate dehydrogenase (GAPD; a loading control). The expression levels of matrilysin were quantified by densitometric scanning (bar graph).

View larger version (125K): [in a new window]   Figure 3. Bacterially induced matrilysin expression colocalizes with lysosomal structures in cultured epithelial cells. Untreated control (A, C, E) and F. necrophorum (AHN 12454)-treated (B, D, F) periodontal ligament epithelial cells were simultaneously immunostained for matrilysin and lysosomal membrane protein h-lamp-1, followed by two fluorescent secondary antibodies Alexa-488 and Alexa-546. Samples were analyzed by laser confocal microscopy, and optical z-axis sections were recorded. Optical sections—one each from the basal, middle, and apical regions of the cells—were presented together. Red channel (A, B), corresponding to Alexa-546 fluorescence, reveals h-lamp-1 expression, and green channel (C, D), corresponding to Alexa-488 fluorescence, reveals matrilysin expression. Red and green images were merged into an RGB file (E, F) showing colocalization (yellow) of the two signals.

View larger version (63K): [in a new window]   Figure 4. Matrix metalloproteinase (MMP) secretion of F. necrophorum-treated epithelial cells. Confluent cultures of porcine periodontal ligament epithelial cells were exposed to F. necrophorum (AHN 12454) for 20 hrs. Untreated cells served as control. Medium aliquots of 25 µL were analyzed for matrix metalloproteinases, by means of gelatin zymography. Known molecular sizes of proMMP-9, -2, and –7 are indicated.