Donald C. Yu

Dentin Matrix Protein-1 Activates Dental Pulp Fibroblasts

INTRODUCTION Dentin matrix protein-1 (DMP-1) is involved in the mineralization of hard dental tissues. DMP-1 is localized in several soft tissues, but its role is unclear. METHODS Human inflamed dental pulps were collected from the endodontic clinic and human normal pulps from impacted teeth. Dental pulp cells from 8 subjects were explanted to test the effect of DMP-1 on interleukin-6 (IL-6) and IL-8 production by using enzyme-linked immunosorbent assay. RESULTS DMP-1 was localized in pulp inflammation by using immunohistochemistry but was not present in impacted root pulps. Wherever found, areas of calcification were positively stained against DMP-1, suggesting its possible involvement in pulp inflammation and in pathologic calcification. To test this hypothesis, primary human pulp fibroblasts were cultured. The fibroblasts were identified on the basis of their morphology, immunoreactivity against vimentin and collagen 1a1 by immunofluorescence and negative staining to CD45, CD34, and cytokeratin by flow cytometry. DMP-1 (10 ng/mL) stimulated the production of IL-6 and IL-8 from pulp fibroblasts. DMP-1 showed an additive effect with lipopolysaccharide in IL-6 and IL-8 production. Inhibition of the p38 mitogen-activated protein kinase pathway blocked the proinflammatory effect of DMP-1 on pulp fibroblasts. CONCLUSIONS Our data indicate that DMP-1 might participate in the development of inflammatory changes in the dental pulp. DMP-1 inhibition might be a new therapeutic strategy to target pulp inflammation and pathologic calcification.

Osteocalcin Expression in Pulp Inflammation

INTRODUCTION Dental pulp inflammation and repair are closely related. Osteocalcin (OCN), a glycoprotein present in dentin matrix, is expressed by odontoblasts. Although OCN is considered a reparative molecule inside the dental pulp, it is not clear if it is involved in pulpal inflammation. The objective of this study was to localize OCN in reversible and irreversible pulpitis and to describe its possible function in inflammation. METHODS Pulp tissues in the form of reversible and irreversible pulpitis were collected from the endodontic clinic. Those from impacted teeth were used as controls. Immunohistochemistry was used to localize OCN. Samples were analyzed for OCN and inflammatory mediator expression using multiplex assay. RESULTS OCN in inflamed tissues was localized in cells and matrix around calcification areas and in cells around blood vessels but not in normal tissues. The plex assay (Bio-Plex 200, Bio-Rad Laboratories Ltd, Mississauga, ON, Canada) showed OCN expression in reversible pulpitis significantly higher than in irreversible pulpitis, and both were significantly higher than in the controls. A panel of inflammatory mediators showed an increase in reversible and irreversible pulpitis. Another panel was decreased in both stages compared with the controls. OCN expression in reversible pulpitis was positively correlated to the expression of vascular endothelial growth factor, fibroblast growth factor, macrophage inflammatory protein-1β, monocyte-derived chemokine, monocyte chemoattractant protein-1, interleukin (IL)-17, and soluble IL-2 receptor α and negatively correlated to that of IL-1α, IL-1β, IL-8, granulocyte macrophage colony-stimulating factor, and macrophage inflammatory protein-1α. CONCLUSIONS Profound understanding of the pulp inflammatory process would lead to new molecular treatment strategies. Our data indicate that OCN expression in reversible pulpitis is associated with angiogenic markers, suggesting its potential use in regenerative treatment.

Effects of Calcitonin, Calcitonin Gene-Related Peptide, Human Recombinant Bone Morphogenetic Protein-2, and Parathyroid Hormone–Related Protein on Endodontically Treated Ferret Canines

INTRODUCTION The purpose of this study was to determine whether human recombinant bone morphogenetic protein-2 (rhBMP-2), calcitonin gene-related peptide (CGRP), calcitonin (CT), or parathyroid hormone-related protein (PTHrP) promoted reparative tertiary dentin or osteodentin formation in ferret canines. METHODS Ferrets had up to 4 pulpotomies performed under anesthesia. All pulps had sterile absorbable sponge of a standard size placed in contact with the pulp. The sponge contained sterile saline, rhBMP-2, CGRP, CT, or PTHrP. The opening was filled with an intermediate restorative material. After 6 weeks, the ferrets were anesthetized, and the pulpotomized teeth were extracted. The canines were fixed, decalcified, sectioned, and stained with hematoxylin-eosin. Sections were selected from the area of the opening, and the amount of reparative tertiary dentin and osteodentin was measured by using a digitizer. RESULTS Analysis of the photomicrographs showed that rhBMP-2 induced 0.58 +/- 0.19 mm(2) osteodentin and 0.56 +/- 0.18 mm(2) tertiary dentin. CGRP induced 0.46 +/- 0.05 mm(2) osteodentin and 0.38 +/- 0.04 mm(2) tertiary dentin. The amount of rhBMP-2-induced and CGRP-induced osteodentin and tertiary dentin was significantly (P < .001) more than that found in the sterile saline-treated teeth (0.29 +/- 0.03 mm(2) osteodentin and 0.14 +/- 0.03 mm(2) tertiary dentin) or CT (0.2 +/- 0.06 mm(2) osteodentin and 0.16 +/- 0.05 mm(2) tertiary dentin; P < .01). PTHrP significantly (P < .05) reduced the amount of osteodentin (0.17 +/- 0.02 mm(2)) observed in the saline-treated teeth but was not significantly different in the amount of tertiary dentin observed. CONCLUSIONS RhBMP-2 and CGRP promoted more pulpal healing than either CT or PTHrP.

Eosinophils in human oral squamous carcinoma; role of prostaglandin D2

Eosinophils are often predominant inflammatory leukocytes infiltrating oral squamous carcinoma (OSC) sites. Prostaglandins are secreted by oral carcinomas and may be involved in eosinophil infiltration. The objective of this study was to determine the factors contributing to eosinophil migration and potential anti-neoplastic effects on OSC. Eosinophil degranulation was evaluated by measuring release of eosinophil peroxidase (EPO). Eosinophil chemotaxis towards OSC cells was assessed using artificial basement membrane. Eosinophil infiltration was prominent within the tissue surrounding the OSC tumor mass. We observed growth inhibition of the OSC cell line, SCC-9, during co-culture with human eosinophils, in vitro, which correlated with EPO activity that possesses growth inhibitory activity. The PGD2 synthase inhibitor, HQL-79, abrogated migration towards SCC-9. Our data suggest that OSC-derived PGD2 may play an important role via CRTH2 (the PGD2 receptor on eosinophils) in eosinophil recruitment and subsequent anti-tumor activity through the action of eosinophil cationic proteins.

The significance of locating and filling the canal isthmus in multiple root canal systems. A scanning electron microscopy study of the mesiobuccal root of maxillary first permanent molars

The mesiobuccal (MB) roots of 50 randomly selected maxillary first permanent molars were examined to evaluate the different configurations of canal isthmus and their incidences. Sections of the roots at 3.4, and 5 mm from the apex were prepared, acid etched, washed and dried. The apical side of each section was sputter coated with gold, examined by a Hitachi S-2500 scanning electron microscope and photographed. 36% of the mesiobuccal roots had one canal, whereas 64% had two canals. In the roots with two canals 31.25% contained either a complete isthmus, or accessory canals between the two main canals 31.25% showed partial isthmus formation. The significance of the different configurations is discussed. Our present findings indicate that the mesiobuccal roots of maxillary first permanent molars exhibit complex anatomy. Prudent judgement is essential in the canal isthmus preparation so that operators must provide meticulous skill and care to the patients.