In situ study of the gelatinase activity in demineralized dentin from rat molar teeth.
Matrix metalloproteinases (MMPs) in dentin are believed to participate in various physiological and pathological events in coronal dentin, but their exact source and location is not clear. The purpose of this study was to evaluate the activity of gelatinases in decalcified rat molars crowns by in situ zymography. Hemi-mandibles of five male Wistar rats were fixed in paraformaldehyde, decalcified in EDTA and glycerol solution and embedded in paraffin. Sections from the region of molar teeth were incubated with or without DQ gelatin in 50mM Tris-CaCl2 at 37°C for 2h and observed by means of confocal microscopy. Gelatinolytic activity was observed throughout the coronal dentin with varying intensities in different locations. High gelatinase activity was observed in the dentinal tubules, dentin-enamel junction (DEJ) and predentin, and it was weaker and less uniform in the intertubular dentin. This study shows that the location of gelatinase and relative activity can be detected by means of in situ zymography and confocal microcopy, and this methodology may provide a useful tool in studies on the role of gelatinases in tooth development, maturation and in pathological conditions.
Department of Morphology, Division of Histology, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil.
Juliana Isabelita Cyrino
JI
Pessoa
Gustavo Narvaes
GN
Guimarães
Naiana Viana
NV
Viola
Wander José
WJ
da Silva
Ana Paula
AP
de Souza
Leo
L
Tjäderhane
Sergio Roberto
SR
Line
Marcelo Rocha
MR
Marques
D000818:Animals; D003804:Dentin / Q000201:enzymology* / Q000378:metabolism; D004789:Enzyme Activation; D018093:Gelatinases / Q000378:metabolism*; D008297:Male; D008963:Molar / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017001:Tooth Demineralization / Q000201:enzymology* / Q000378:metabolism
D018093:Gelatinases