A Study In Situ Of The Effect Of Metallo- And Serine Proteinase Inhibitors On The Birefringence Of The Secretory Stage Enamel Organic Extracellular Matrix.

dc.creatordo Espirito Santo, A R
dc.creatorMarques, M R
dc.creatorLine, S R P
dc.date2011-Apr
dc.date2015-11-27T13:20:58Z
dc.date2015-11-27T13:20:58Z
dc.date.accessioned2018-03-29T01:12:27Z
dc.date.available2018-03-29T01:12:27Z
dc.identifierBiotechnic & Histochemistry : Official Publication Of The Biological Stain Commission. v. 86, n. 2, p. 108-14, 2011-Apr.
dc.identifier1473-7760
dc.identifier10.3109/10520290903472407
dc.identifierhttp://www.ncbi.nlm.nih.gov/pubmed/20109097
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/199254
dc.identifier20109097
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1299487
dc.descriptionDental enamel formation occurs extracellularly and establishment of an ordered enamel organic extracellular matrix (ECM) seems to be crucial for proper construction of the enamel mineral phase. Polarizing microscopy shows that the ordered supramolecular structure of the secretory stage enamel organic ECM exhibits strong birefringence. We reported earlier that this birefringence is lost in unfixed specimens, probably due to extensive proteolytic cleavage of enamel proteins. Therefore, we investigated the association between enamel proteinase activities by analyzing the effects of metallo- and serine proteinase inhibitors in situ on the birefringence of the secretory stage enamel organic ECM. Male rats were used in the present study. After sacrifice, distal 10 mm fragments of upper incisors were removed and immersed for 15 h under continuous shaking at 37°C in one of the following solutions: 1) 10 mM Tris, pH 8.0; 150 mM NaCl (negative control, n = 8); 2) 2% paraformaldehyde and 0.5% glutaraldehyde in 0.2 M phosphate-buffered saline (PBS), pH 7.2 (positive control, n = 5); 3) 10 mM Tris, pH 8.0; 150 mM NaCl; 2 mM 1,10-phenanthroline (n = 9); 4) 10 mM Tris, pH 8.0; 150 mM NaCl; 2 mM phenylmethyl-sulfonyl fluoride (PMSF) (n = 8); 5) 10 mM Tris, pH 8.0; 150 mM NaCl; 2 mM 1,10-phenanthroline; 2 mM PMSF (n = 9). Samples then were immersed in fixative solution for 24 h and processed to obtain 5 μm thick longitudinal sections of the secretory stage enamel organic ECM. The sections were immersed in 80% glycerin for 30 min and analyzed by transmitted polarizing light microscopy. 1,10-Phenanthroline (inhibitor of metalloproteinases) and 1,10-phenanthroline + PMSF (inhibitor of serine proteinases) clearly prevented a decrease in the optical retardation of birefringence brightness from the tissue. PMSF alone promoted a slight preservation of the birefringence exhibited by the secretory stage enamel organic ECM. Rapid loss of birefringence in secretory stage enamel organic ECM that is not fixed immediately is caused by enamel proteinases and the activity of metalloproteinases seems to lead to preliminary degradation of the enamel organic ECM, which in turn facilitates subsequent serine proteinase activity.
dc.description86
dc.description108-14
dc.languageeng
dc.relationBiotechnic & Histochemistry : Official Publication Of The Biological Stain Commission
dc.relationBiotech Histochem
dc.rightsfechado
dc.rights
dc.sourcePubMed
dc.subjectAnimals
dc.subjectBirefringence
dc.subjectCollagenases
dc.subjectDental Enamel
dc.subjectExtracellular Matrix
dc.subjectMale
dc.subjectMatrix Metalloproteinase Inhibitors
dc.subjectMicroscopy, Polarization
dc.subjectPhenanthrolines
dc.subjectPhenylmethylsulfonyl Fluoride
dc.subjectRats
dc.subjectRats, Wistar
dc.subjectSerine Proteases
dc.subjectSerine Proteinase Inhibitors
dc.titleA Study In Situ Of The Effect Of Metallo- And Serine Proteinase Inhibitors On The Birefringence Of The Secretory Stage Enamel Organic Extracellular Matrix.
dc.typeArtículos de revistas


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