bluuebunny/crossref_metadata_2025 huggingface dataset:
The Activin-binding Protein Follistatin is Expressed in Developing Murine Molar and Induces Odontoblast-like Cell Differentiation in vitro
Authors: Heikinheimo K. , Begue-Kirn C. , Ritvos O. , Tuuri T. , Ruch J.V.
Published: January 2009

It has recently been shown that mice deficient in activin-βA subunits and follistatin exhibit major defects in dentition. To increase understanding of the roles played by these molecules during tooth development, we determined the temporospatial expression of activin-βA subunit and follistatin messenger RNA and their corresponding proteins in developing murine molars (between day E 14 and 2 days after birth). The effects of recombinant human activin A and its binding protein follistatin on odontoblast differentiation were also studied in cultures of dental papillae (DP) isolated from the mandibular first molars of E-17-day mice. In situ hybridization indicated that transcripts for activin-βA subunit were abundant in pre-odontoblasts at the tips of forming cusps prior to odontoblast terminal differentiation, and transcripts for follistatin in overlying inner enamel epithelial cells (pre-ameloblasts). Pre-odontoblasts were also weakly immunoreactive in relation to activin-βA subunit, pre-ameloblasts in relation to follistatin. When follistatin was added at different concentrations to a DP culture model (2-14 nmol/DP) together with heparin at constant concentration, differentiation of odontoblast-like cells was induced, as evidenced by polarization and deposition of extracellular matrix in vitro, to extents depending on the follistatin concentration. In contrast, the addition of activin A (2 nmol/DP) had no effect on the differentiation parameters studied. These findings suggest that the activin-follistatin system regulates odontoblast differentiation during tooth development. In particular, we suggest that binding of endogenous activin A by follistatin may allow odontoblast terminal differentiation to occur.
Changes in the Shape and Orientation of Periodontal Ligament Fibroblasts in the Continuously Erupting Rat Incisor Following Removal of the Occlusal Load
Authors: Weinreb M. , Gal D. , Weinreb M.M. , Pitaru S.
Published: January 2009

One of the main theories which attempts to explain the phenomenon of tooth eruption suggests that periodontal ligament (PDL) fibroblasts move actively and pull the tooth with them out of its socket. To find further support for this theory, we determined the changes in the shape and orientation of PDL fibroblasts induced by a transition from impeded to unimpeded eruption. We measured nuclear area, elongation (length-to-width ratio), and orientation (angulation in relation to the eruption axis) of PDL fibroblasts in impeded (functionally loaded) and unimpeded (hypoloaded) rat incisors. The mean cross-sectional nuclear area did not differ between fibroblasts in the two groups. In contrast, unimpeded eruption resulted in a marked increase in the mean nuclear elongation (from about 2 to 2.56) and a significant increase in the mean nuclear orientation (from 25.6 to 14.0 degrees). Bivariate analysis suggested that these changes occurred in the same cells. Analysis of nuclear elongation and orientation at various distances from the cementum toward the alveolar bone revealed a profile of both parameters, such that cells located 20 to 80 μm away from the cemental surface were more elongated and more frequently oriented toward the eruption axis, while cells at 0 to 20 and 80 to 100 μm were more round/oval and had a greater angulation with the eruption axis. These findings, together with other observations of changes in cell number, number of microtubules, and migration velocity which occur on the shift to unimpeded eruption, support the theory of active movement of PDL fibroblasts as an important component of tooth eruption.
A Three-dimensional Finite Element Model of Prismatic Enamel: A Re-appraisal of the Data on the Young's Modulus of Enamel
Authors: Spears I.R.
Published: January 2009

The inconsistencies of published data on the Young's modulus of dental enamel, the parameter used to quantify stiffness, have, for a long time, restricted our understanding of the biomechanical behavior of teeth. With the use of modeling techniques, the aim of this paper is to investigate which of the data may be more reliable. In this way, the possible causes of the discrepancies in data will be addressed. Two different structural levels are considered within the model. At an ultrastructural (i.e., crystalline) level, the model considers enamel to behave as a simple composite, being made up of long, parallel crystals held together by an organic matrix. At this level, the stiffness of enamel is predicted by simple composite theory, and the model indicates that stiffness is dependent on chemical composition and crystal orientation. At a microstructural (i.e., prismatic) level, the model considers enamel to behave as a hierarchical composite, being made up of prisms, in which the crystal orientation is heterogeneous. At this level, the stiffness of enamel is predicted by finite element stress analysis, and values of predicted stiffness are found to be dependent on both chemical composition and prism orientation. Within a realistic compositional range, predicted values of Young's modulus along the direction of prisms are comparable with the corresponding experimental values of 77.9 ± 4.8 GPa obtained by Craig et al. (1961) and 73 GPa obtained by Gilmore et al. (1970), but not with those low values of 9.65 ± 3.45 obtained by Stanford et al. (1960). Predictions of Young's modulus values across the direction of prisms are also made, and the model is less stiff in this direction. These findings indicate that human prismatic enamel is almost certainly anisotropic with respect to stiffness.
Enamel Subsurface Damage Due to Tooth Preparation with Diamonds
Authors: Xu H.H.K. , Kelly J.R. , Jahanmir S. , Thompson V.P. , Rekow E.D.
Published: January 2009

In clinical tooth preparation with diamond burs, sharp diamond particles indent and scratch the enamel, causing material removal. Such operations may produce subsurface damage in enamel. However, little information is available on the mechanisms and the extent of subsurface damage in enamel produced during clinical tooth preparation. The aim of this study, therefore, was to investigate the mechanisms of subsurface damage produced in enamel during tooth preparation by means of diamond burs, and to examine the dependence of such damage on enamel rod orientation, diamond particle size, and removal rate. Subsurface damage was evaluated by a bonded-interface technique. Tooth preparation was carried out on two enamel rod orientations, with four clinical diamond burs (coarse, medium, fine, and superfine) used in a dental handpiece. The results of this study showed that subsurface damage in enamel took the form of median-type cracks and distributed microcracks, extending preferentially along the boundaries between the enamel rods. Microcracks within individual enamel rods were also observed. The median-type cracks were significantly longer in the direction parallel to the enamel rods than perpendicular to the rods. Preparation with the coarse diamond bur produced cracks as deep as 84 ± 30 μm in enamel. Finishing with fine diamond burs was effective in crack removal. The crack lengths in enamel were not significantly different when the removal rate was varied. Based on these results, it is concluded that subsurface damage in enamel induced by tooth preparation takes the form of median-type cracks as well as inter- and intra-rod microcracks, and that the lengths of these cracks are sensitive to diamond particle size and enamel rod orientation, but insensitive to removal rate.
Identification, Partial Characterization, and Distribution of Versican and Link Protein in Bovine Dental Pulp
Authors: Yamauchi S. , Cheng H. , Neame P. , Caterson B. , Yamauchi M.
Published: January 2009

The dynamics of changes in the cellularity and extracellular matrix composition of dental pulp varies considerably during tooth development and maturation. In this paper, we studied matrix proteoglycans where we hypothesized that they played important roles in structural, spatial, and transport aspects of pulpal development and maintenance. The pulpal tissue was collected from partially erupted bovine incisors, pulverized, and then extracted with 6 M guanidine-HCl. The extract was subjected to anion column chromatography (DEAE-8HR), and the fractions collected were screened by dot-blot immunoassay by means of monoclonal antibodies generated against 4- and 6-sulfated chondroitin sulfate isomers, and keratan sulfate, 2-B-6, 3-B-3, and 5-D-4, respectively. The chondroitin-6-sulfate was the major glycosaminoglycan species and occurred as a large-molecular-weight proteoglycan (> 500 kDa). After further purification, it was subjected to agarose/acrylamide composite gel electrophoresis, and it migrated as a single band stained with Stains-All. The band was immunopositive against antibody 3-B-3 by Western blot analysis. The partial amino acid sequence analyses of the core protein clearly indicated this molecule to be versican. The presence of link protein was also confirmed by Western blot analysis with an anti-link protein monoclonal antibody, 8-A-4. Furthermore, immunohistochemical study indicated that the distributions of versican and link protein coincide in the dental pulp and are enriched in the peripheral area of the tissue just beneath the odontoblast layer. Since the dental pulp contains hyaluronan, versican may bind to hyaluronan via its hyaluronan-binding domain, where this association is stabilized by link protein. This complex, then, could form large hydrated proteoglycan aggregates that fill the extracellular space, support odontoblasts, and/or facilitate the transport function of metabolites and nutrients within the tissue.
Impact Velocities of the Teeth after a Sudden Unloading at Various Initial Bite Forces, Degrees of Mouth Opening, and Distances of Travel
Authors: Nagashima T. , Slager G.E.C. , Otten E. , Broekhuijsen M.L. , van Willigen J.D.
Published: January 2009

A potentially dangerous situation arises when an individual bites on hard and brittle food which suddenly breaks, since the impact velocity of the lower teeth onto the upper teeth after the food is broken can be high and may cause dental damage. The present experiments were designed to study the magnitude of the impact velocity after a sudden unloading at various initial bite forces, degrees of mouth opening, and distances of travel. Subjects were asked to perform a static biting task during which the resistance to the bite was suddenly removed. The upward mandible movement was arrested after a certain distance. The velocity of the lower teeth at impact was calculated just before the mandible came to a standstill in combinations of 4 different bite forces (100, 80, 60, and 40 N), 4 different initial degrees of mouth opening (33.5, 30.5, 27.5, and 24.5 mm), and 3 different distances of travel of the mandible (4.5, 3.0, and 1.5 mm). We found that the bite force rapidly declined after the unloading, resulting in a small impact velocity of the lower front teeth. This impact velocity largely depended on the magnitude of the initial bite force and the distance traveled; it was barely sensitive to variations in degree of initial mouth opening. The maximal velocity of the lower teeth was 0.43 m/s (at an initial bite force of 100 N). This maximum was reached after a distance of travel of about 4 mm in 12 ms. The data suggest that the rapid decline in bite force coupled with a limitation of impact velocity is due to the force-velocity properties of the active jaw muscles and is not caused by neural control.
Viscoelastic Properties of the Pig Temporomandibular Joint Articular Soft Tissues of the Condyle and Disc
Authors: Kuboki T. , Shinoda M. , Orsini M.G. , Yamashita A.
Published: January 2009

It has been suggested that a sustained loading condition such as clenching could compress the temporomandibular joint (TMJ) articular soft tissues. However, there is still no clear understanding of how the TM joint articular tissues respond under compression. To answer this question, we performed in vitro indentation tests on fresh articular discs and cartilage-bone systems of the condyles of 10 Yorkshire pigs (aged 7 months) using a self-developed indentation tester. The indenter was 5 mm in diameter and was controlled by means of a computer-aided feedback mechanism. Bilateral condyles from the same mandible were uniformly prepared; one was used for measurements under sustained compression (SC) and the other for measurements under intermittent compression (IC). The displacements of the indenter induced by a SC of 10, 20, and 30 Newtons (N, units of force) for 10 min and by an IC, also of 10, 20, and 30 N, with one-second duration and two-second intervals for 10 min were measured by means of a displacement sensor with a resolution of 0.001 mm. From these data, the indentation curves of the articular discs and the cartilage-bone systems were calculated. Both the disc and the articular cartilage showed characteristic displacement vs. time curves-namely, an instantaneous deformation upon load application, followed by a time-dependent creep phase of asymptotically increasing deformation under constant load. However, the indentation curves of the two tissues were not identical: The deformation of the articular cartilage was dose-dependent, but that of the disc was not. Moreover, the articular cartilage deformed significantly less under IC than under SC. This difference was not found in the disc. It can be concluded that both the disc and the articular cartilage of the pig temporomandibular joint have viscoelastic properties against compression; however, the disc is stiffer than the articular cartilage.
The Influence of the Amalgam Alloy on the Survival of Amalgam Restorations: A Secondary Analysis of Multiple Controlled Clinical Trials
Authors: Letzel H. , Van 'T Hof M.A. , Marshall G.W. , Marshall S.J.
Published: January 2009

Data from 14 independent controlled clinical trials on the oral behavior of Classes 1 and 2 amalgam restorations, with a follow-up between five and 15 years, were re-evaluated by secondary analysis for the influence of alloy composition on the survival of amalgam restorations. For the analysis, 3119 restorations were available, which were made from 24 different alloys by a group of seven operators. The alloys were divided into four groups according to their zinc content (zinc-containing and zinc-free) and their copper content (conventional and high-copper). During the follow-up of the trials, the restorations were annually assessed for failures, which were classified as to (1) restoration-, (2) restorative process-, and (3) patient-related reasons. With the restoration-related failures, survival functions of the restorations were estimated by alloy and alloy group. The total number of failed restorations was 481, of which 77% were restoration-related and 14% process-related. Eighty percent of the restoration-related failures were due to some form of fracture of the amalgam. Restorations of conventional zinc-free alloys had the shortest survival. After 13 years, only 25% survived. Zinc and a high copper content had an equally favorable influence on the survival rate, which was 70% after 13 years when either was present. The highest survival rates were of restorations of zinc-containing high-copper alloys: 85% after 13 years. The zinc and copper contents of the alloy contributed to the corrosion resistance of the amalgams, which in turn influenced the survival of the restoration. The current ISO Standard 1559 on alloys for dental amalgam should be modified to account for these factors that influence the survival of amalgam restorations.
Profile of Cytokine mRNA Expression in Chronic Adult Periodontitis
Authors: Roberts EA , McCaffery K.A. , Michalek S.M.
Published: January 2009

Chronic inflammation induced by bacteria often leads to host-mediated destruction of tissues adjacent to the sites of microbial insult. The chronic inflammatory process of adult periodontitis results in the destruction of supporting osseous and connective tissues of the teeth. We hypothesized that virulence factors of periodontal pathogens such as lipopolysaccharide stimulate inflammatory cytokine expression by mononuclear cells of the host which contribute to disease development. In this study, to elucidate the role of these cytokines in chronic adult periodontitis, we tested whether the prevalence of mRNA for inflammatory cytokines generally associated with mononuclear phagocytes was higher in diseased than in healthy gingival tissue. Gingival mononuclear cells or whole gingival biopsies from 32 adult periodontitis patients and five healthy individuals used as controls were evaluated for inflammatory cytokine mRNA expression by reverse-transcription polymerase chain-reaction (RT-PCR) procedures. The cytokines assessed included IL-1α, IL-1β, IL-lra, IL-6, IL-8, IL-12, IL-13, TNF-a, TGF-β, and IFN-γ. The monocyte/macrophage lipopolysaccharide (LPS) receptor CD14 was also assessed. Results showed that TNF-α mRNA was present significantly more frequently in diseased than in healthy biopsies, whereas IL-la, IL-1β, and IL-lra mRNA were found in most (from 80 to 100%) healthy tissues. Message for CD14 was present in both healthy and diseased tissue samples (100%). This study provides evidence for a major role of TNF-a in chronic adult periodontitis. Moreover, our results suggest that the mononuclear cells derived from periodontal tissues have the capacity to respond to components of periodontal pathogens and express both pro- and anti-inflammatory cytokines in these tissues.
Prevalence and Distribution of Six Capsular Serotypes of Porphyromonas gingivalis in Periodontitis Patients
Authors: Laine M.L. , Appelmelk B.J. , van Winkelhoff A.J.
Published: January 2009

Previous reports have described six serotypes based on K antigens in Porphyromonas gingivalis strains. The purpose of the present study was to investigate the prevalence and distribution of these serotypes in 185 patients with P. gingivalis-associated periodontitis. Polyclonal rabbit antisera, raised against each of the different type strains, were used in double-immunodiffusion and immunoelectrophoresis assays. In addition, a subset of 76 strains was investigated for the presence of capsular structures by means of the India ink and Bruce White staining techniques. These strains were also tested for auto-aggregation in phosphate-buffered saline (PBS). All six K serotypes were present in the study sample. In total, 84 (45.4%) patients were colonized with a K-typeable P. gingivalis strain with a predominance of types K5 (12%) and K6 (23.2%). A correlation was found between arbitrary age categories and the prevalence of currently known K serotypes, which were found in 60% of patients aged 12 to 30 years, in 49% of patients aged 31 to 50, and in 25% of patients aged 51 to 70 years. In the subset of 76 P. gingivalis strains, 32 (42.1%) were K-typeable. Fifty-three strains (69.7%) showed microscopic evidence of encapsulation, suggesting the existence of K serotypes other than K1 to K6. Twenty-one strains (27.6%) auto-aggregated in PBS and were not K-typeable, nor did they show any evidence of encapsulation. It was concluded that the majority of clinical P. gingivalis isolates is encapsulated and that encapsulation is associated with the presence of a K antigen. Auto-aggregation seems to be associated with the absence of a capsular structure and, consequently, the absence of a K antigen.
Arrest of Root Surface Caries in situ
Authors: Nyvad B. , Ten Cate J.M. , Fejerskov O.
Published: January 2009

This study tests the hypothesis that daily oral hygiene combined with topical fluoride arrests active root-surface caries lesions without changing the mineral content of the lesions. Therefore, changes in mineral content and distribution were studied in root surfaces during caries lesion development and subsequent arrest of lesion progression in situ. In 18 subjects, lesions were developed during 3 months in sound root-surface specimens inserted into lower partial dentures. After 3 months, ground sections were prepared from each lesion prior to re-insertion of the specimens into the dentures. In addition, one sound root specimen was added per subject. During the following 3 months, half of the subjects cleaned both sound and carious specimens once a day with an 1100-ppm fluoride toothpaste, and the specimens were treated twice with 2% NaF for 2 min in situ. The other half of the subjects continued the experiment without cleaning. During the initial three-month period, all specimens developed subsurface lesions extending 187 to 583 μm into the dentin. Lesion depth increased somewhat in both experimental groups during the following 3 months (P ≥ 0.1). There was a non-significant increase in mineral loss in the plaque-covered specimens (P = 0.08). However, the total mineral content of specimens subjected to plaque removal and topical fluoride did not change. This treatment resulted in an increased mineral content in the surface layer (P < 0.01) and formation of a zone of higher mineral content within the body of the lesion. The sound root surfaces which had been cleaned for a three-month period showed mineral uptake in the surface layer, occasionally associated with subsurface demineralization extending 20 to 70 μm into the tissue. The mineral loss of these specimens was significantly smaller than that of plaque-covered surfaces (P < 0.001). It is concluded that daily plaque removal and topical fluoride use influence the distribution of mineral in sound and carious root surfaces and may arrest lesion progression without affecting the total mineral content.
Effects of Various Resin Composite (Co)monomers and Extracts on Two Caries-associated Micro-organisms in vitro
Authors: Hansel C. , Leyhausen G. , Mai U.E.H. , Geurtsen W.
Published: January 2009

Previous studies have shown that extracts of various filling materials, e.g., resin composites, may influence the growth of cariogenic micro-organisms. The purpose of this study was to examine the effects of important resin composite (co)monomers (Bis-GMA, UDMA, TEGDMA, EGDMA) and extracts of two commercial dental composites with similar composition (composite A, Arabesk™; composite S, Superlux™) on the growth of the two cariogenic bacterial pathogens Streptococcus sobrinus and Lactobacillus acidophilus. It was found that neither the monomers Bis-GMA and UDMA, nor the comonomer EGDMA, nor the extract of composite A influenced the growth of S. sobrinus in the log phase. The comonomer TEGDMA and the extract of composite S were found to stimulate growth in the log phase, but this stimulation was not statistically significant. However, EGDMA, TEGDMA, and the extract of composite S did stimulate the total growth of S. sobrinus. In the assays with L. acidophilus, Bis-GMA, UDMA, and the extract of composite A inhibited the growth in the log phase, whereas TEGDMA stimulated it. Furthermore, EGDMA, TEGDMA, and the extract of composite S stimulated the biomass production of L. acidophilus. We conclude from our results that a release of EGDMA and TEGDMA from resin composites should be avoided due to their growth-stimulating effects on the caries-associated micro-organisms S. sobrinus and L. acidophilus.
Potassium and Chloride Channels in Freshly Isolated Rat Odontoblasts
Authors: Guo L. , Davidson R.M.
Published: January 2009

It has been suggested that understanding the physiological properties of odontoblasts may be important in understanding the mechanisms underlying both metabolic and transductive processes in dental pulp. Because ion flux(es) may play a critical role in these events, it is of particular interest to understand ionic mechanisms in odontoblast cells. Thus, the aim of this study was to use patch-clamp recording techniques to examine the properties of resident ion channels in freshly dissociated odontoblasts. In recordings made in potassium-rich solutions, cells displayed at least three distinct channel amplitudes, with conductances of 130 ± 18 pS, 52 ± 4 pS, and 25 ± 2 pS, respectively. Channel activity persisted in the presence of potassium salts of impermeant anions, and could be abolished by barium, a non-specific potassium channel blocker. In addition to the potassium conductances, we saw two separate anion channels in the odontoblast membrane. These channels were predominantly chloride-selective, weakly permeable to both acetate and aspartate, and had conductances of 391 ± 64 pS and 24 ± 3 pS. While questions remain regarding the functional role of these and other ion channels that presumably reside in the odontoblast membrane, our results demonstrate that it is possible to study ionic mechanisms of the odontoblast at the level of the single cell.
Swallowing-related Perihypoglossal Neurons Projecting to Hypoglossal Motoneurons in the Cat
Authors: Ono T. , Ishiwata Y. , Kuroda T. , Nakamura Y.
Published: January 2009

Although previous studies have examined the functional role of the neurons in the area ventrolateral to the hypoglossal nucleus (perihypoglossal neurons) in the trigemino-hypoglossal reflex, no convincing evidence for the direct connection from the perihypoglossal neurons to the hypoglossal motoneurons has yet been provided. In addition, the role of the perihypoglossal neurons in swallowing has not been studied. The purpose of this study was to investigate (1) the input-output relationship of the perihypoglossal neurons and (2) whether the afferent feedback was essential for their swallowing-related activity in chloralose-anesthetized cats. Before and after the cats were paralyzed, single-unit activities were recorded extracellularly from 30 perihypoglossal neurons during swallowing elicited by electrical stimulation of the superior laryngeal nerve. These perihypoglossal neurons responded with spike potentials after short latencies to stimulation of the inferior alveolar and hypoglossal nerves. The neurons also responded with spike potentials to single shocks applied to the superior laryngeal nerve, but were activated transiently at the initial phase of repetitive stimulation of the nerve and kept silent until the occurrence of swallowing before and after the animal was paralyzed. They showed burst activities in coincidence with swallowing. Averaging of intracellular potentials of a hypoglossal motoneuron by simultaneously recorded extracellular spikes of a perihypoglossal neuron revealed monosynaptic inhibitory post-synaptic potentials. We conclude that, in the region ventrolateral to the hypoglossal nucleus, there are neurons which relay trigeminal, hypoglossal, and vagal afferents. Furthermore, some of these perihypoglossal neurons are inhibitory hypoglossal premotor neurons that are involved in the central programming of swallowing.
"In vitro" Dissolution of Coral in Peritoneal or Fibroblast Cell Cultures
Authors: Fricain J.-Ch. , Bareille R. , Rouais F. , Basse-Cathalinat B. , Dupuy B.
Published: January 2009

Previous studies have shown that in vivo coral resorption involves a biphasic process: First, the edges of the coral block become powdery, then extracellular fluid and phagocytosis contribute to the dissolution of the crystals. The authors examined some types of cells that could be involved in phagocytosis, particularly the ability of both dermal fibroblasts and mouse-resident peritoneal cells to phagocytose and dissolve coral powder "in vitro". Radioactive coral was incubated for 24, 48, or 72 hrs with cells in the presence or absence of cytochalasin B (a phagocytic inhibitor) or chloroquine (a lysosomotropic agent). Furthermore, to specify the role of crystal cell contacts in the solubilization process, they incubated radioactive coral in conditioned media (obtained from two-day human fibroblastic or macrophagic cell culture in the presence or absence of non-radioactive coral) or at a distance from the cells using culture inserts. Measurements of the radioactivity in the different supernatants were performed. Transmission electron microscopy was carried out on the cells cultivated in the presence or absence of radioactive coral. The data suggest that both fibroblasts and macrophages dissolve the coral, and that the intracellular degradation in phagolysosomes is one of the mechanisms explaining coral powder dissolution.
Mercury Concentrations in Urine and Whole Blood Associated with Amalgam Exposure in a US Military Population
Authors: Kingman A. , Albertini T. , Brown L.J.
Published: January 2009

Minute amounts of mercury vapor are released from dental amalgams. Since mercury vapor is known to be associated with adverse health effects from occupationally exposed persons, questions regarding the margin of safety for exposure to mercury vapor in the general population continue to be raised. To address this issue, one needs information regarding exposure to mercury vapor from dental amalgam fillings and its possible consequences for health in the general population. The NIDR Amalgam Study is designed to obtain precise information on amalgam exposure and health outcomes for a non-occupationally-exposed population of US adults. One hypothesis was that in a generally healthy population a significant association between amalgam exposure and Hg levels in urine and/or whole blood could be detected. The cohort investigated was an adult military population of 1127 healthy males. Their average age was 52.8 years, and their ages varied from 40 to 78 years. Ninety-five percent of the study participants were white males, and slightly over 50% had some college education. Five percent were edentulous. The dentate participants, on average, had 25 natural teeth, 36.9 decayed or filled surfaces (DFS), and 19.9 surfaces exposed to amalgam, with amalgam exposure varying from 0 to 66 surfaces. Their average total and inorganic urinary mercury concentrations were 3.09 μg/L and 2.88 pg/L. The average whole-blood total and inorganic mercury concentrations were 2.55 ug/L and 0.54 μg/L. Significant correlations were detected between amalgam exposure and the total (r = 0.34, p < 0.001) and inorganic 0.34 (r = 0.34, p < 0.001) urinary mercury concentrations on the original scale. Stronger correlations were found for total (r = 0.44, p < 0.001) and inorganic (r = 0.41, p < 0.001) urinary Hg on the log scale, as well as for creatinine-corrected total (r = 0.43, p < 0.001) and inorganic (r = 0.43, p < 0.001) urine concentrations. In whole blood, statistically significant, but biologically weak, correlations were detected for total (r = 0.09, p = 0.005) and inorganic (r = 0.15, p < 0.001) Hg concentrations, respectively. Based on these cross-sectional data, it is estimated that, on average, each ten-surface increase in amalgam exposure is associated with an increase of 1 ug/L mercury in urine concentration.
Indentation Damage and Mechanical Properties of Human Enamel and Dentin
Authors: Xu H.H.K. , Smith D.T. , Jahanmir S. , Romberg E. , Kelly J.R. , Thompson V.P. , Rekow E.D.
Published: January 2009

Understanding the mechanical properties of human teeth is important to clinical tooth preparation and to the development of "tooth-like" restorative materials. Previous studies have focused on the macroscopic fracture behavior of enamel and dentin. In the present study, we performed indentation studies to understand the microfracture and deformation and the microcrackmicrostructure interactions of teeth. It was hypothesized that crack propagation would be influenced by enamel rods and the dentino-enamel junction (DEJ), and the mechanical properties would be influenced by enamel rod orientation and tooth-to-tooth variation. Twenty-eight human third molars were used for the measurement of hardness, fracture toughness, elastic modulus, and energy absorbed during indentation. We examined the effect of enamel rod orientation by propagating cracks in the occlusal surface, and in the axial section in directions parallel and perpendicular to the occlusal surface. The results showed that the cracks in the enamel axial section were significantly longer in the direction perpendicular to the occlusal surface than parallel. The cracks propagating toward the DEJ were always arrested and unable to penetrate dentin. The fracture toughness of enamel was not single-valued but varied by a factor of three as a function of enamel rod orientation. The elastic modulus of enamel showed a significant difference between the occlusal surface and the axial section. It is concluded that the cracks strongly interact with the DEJ and the enamel rods, and that the mechanical properties of teeth are functions of microstructural orientations; hence, single values of properties ( e.g., a single toughness value or a single modulus value) should not be used without information on microstructural orientation.
Modification of Amino Acid Residues in Carious Dentin Matrix
Authors: Kleter G.A. , Damen J.J.M. , Buijs M.J. , Ten Cate J.M.
Published: January 2009

The Maillard reaction between sugar and protein has been postulated as the cause for the browning and arrestment of caries lesions. This reaction has been implicated as the cause for decreased degradability of collagen in vivo. The aim of the present study was to verify the occurrence of the reaction in vivo. Carious and sound dentin samples were taken from extracted human teeth and analyzed for the fluorescence characteristic of the Maillard reaction and oxidation and, by HPLC, for Maillard products. In addition, physiological cross-links were analyzed by HPLC. Oxidation- and Maillard reaction-related fluore-scence increased in collagenase digests from carious dentin. Advanced Maillard products (carboxymethyllysine and pentosidine) increased, whereas furosine, a marker for the initial reaction, was not observed consistently. This implies no direct addition of sugars to protein, but rather the addi-tion of smaller metabolites and glycoxidation products. In addition, the physiological crosslinks hydroxylysinonorleucine and dihydroxylysinonorleucine decreased in carious dentin. Also for hydroxylysylpyridinoline, a decrease was observed, but not consistently. In conclusion, the caries process modifies amino acids in dentin collagen, which can lead to increased resistance against proteolysis and ulti-mately to caries arrestment.
Protein-to-Protein Interactions: Criteria Defining the Assembly of the Enamel Organic Matrix
Authors: Paine M.L. , Krebsbach P.H. , Chen L.S. , Paine C.T. , Yamada Y. , Deutsch D. , Snead M.L.
Published: January 2009

Enamel crystallites form in a protein matrix located proximal to the ameloblast cell layer. This unique organic extracellular matrix is constructed from structural protein components biosynthesized and secreted by ameloblasts. To date, three distinct classes of enamel matrix proteins have been cloned. These are the amelogenins, tuftelin, and ameloblastin, with recent data implicating ameloblastin gene expression during cementogenesis. The organic enamel extracellular matrix undergoes assembly to provide a three-dimensional array of protein domains that carry out the physiologic function of guiding enamel hydroxyapatite crystallite formation. Using the yeast two-hybrid system, we have surveyed these three known enamel gene products for their ability to direct self-assembly. We measured the capacity of the enamel gene products to direct protein-to-protein interactions, a characteristic of enamel proteins predicated to be required for self-assembly. We provide additional evidence for the self-assembly nature of amelogenin and tuftelin. Ameloblastin self-assembly could not be demonstrated, nor were protein-to-protein interactions observed between ameloblastin and either amelogenin or tuftelin. Within the limits of the yeast two-hybrid assay, these findings constrain the emerging model of enamel matrix assembly by helping to define the limits of enamel matrix protein-protein interactions that are believed to guide enamel mineral crystallite formation.
Gustducin and its Role in Taste
Authors: Spielman A.I.
Published: January 2009

The mechanisms responsible for taste signal transductions are very complex. A key molecule, α-gustducin, a primarily taste-specific G protein α-subunit, was discovered in 1992 and was later found to be Involved in both bitter and sweet taste transduction. A proposed mechanism for α-gustducin involves coupling specific cell-surface receptors with a cyclic nucleotide phosphodiesterase which would open a cyclic nucleotide-suppressible cation channel leading to influx of calcium, and ultimately leading to release of neurotransmitter. Although "knock-out" animals deficient in the α-gustducin gene clearly demonstrate that gustducin is an essential molecule for tasting certain bitter and sweet compounds, the precise role of α-gustducin in bitter and sweet taste is presently unclear, Indeed, there are several other signaling mechanisms in sweet and bitter taste, apparently unrelated to α-gustducin, that increase cyclic AMP or inositol 1,4,5 trisphosphate. Thus, proposed models for α-gustducin and those found by other laboratories may be parallel and interdependent.
Bone Morphogenetic Protein-7 (Osteogenic Protein-1, OP-1) and Tooth Development
Authors: Helder M.N. , Karg H. , Bervoets T.J.M. , Vukicevic S. , Burger E.H. , D'Souza R.N. , Wöltgens J.H.M. , Karsenty G. , Bronckers A.L.J.J.
Published: January 2009

Bone morphogenetic proteins (BMPs) form a family of growth factors originally isolated from extracellular bone matrix that are capable of inducing bone formation ectopically. We studied the expression, tissue localization, and function of BMP-7 (OP-1) during tooth development in rodents. Patterns of BMP-7 gene expression and peptide distribution indicated that BMP-7 was present in dental epithelium during the dental lamina, bud, and cap stages. During the bell stage, BMP-7 mRNA expression and protein distribution shifted from dental epithelium toward the dental mesenchyme. With advancing differentiation of odontoblasts, BMP-7 protein staining in the dental papilla became restricted to the layer of fully functional odontoblasts in the process of depositing (pre)dentin. Secretory-stage ameloblasts exhibited weak immunostaining for BMP-7. A restricted pattern of staining in ameloblasts became apparent in post-secretory stages of amelogenesis. Also, cells of the forming periodontal ligament were immunopositive. Histological analysis of tooth development in neonatal BMP-7-deficient mice did not reveal obvious changes compared with wild-type mice. We conclude that, in developing dental tissues, BMP-7 has distribution and expression patterns similar to those of other BMP members but is not an essential growth factor for tooth development, possibly because of functional redundancy with other BMP members or related growth factors.
Quantitative Analysis of Amelogenin Solubility
Authors: Tan J. , Leung W. , Moradian-Oldak J. , Zeichner-David M. , Fincham A.G.
Published: January 2009

Amelogenins are a group of extracellular enamel matrix proteins which are believed to be involved in the regulation of the size and habits of forming enamel crystals. The aim of this study was to compare the solubility properties of several amelogenins at various pH (from 4.0 to 9.0) at constant ionic strength (IS), and to examine the influence of buffer composition, IS, and divalent metal ions (including Ca2+, Mg2+, and Zn2+) on amelogenin solubility. The solubility of the recombinant murine amelogenin ("rM179") was minimum near its isoelectric point and increased rapidly below and above, regardless of buffer composition. A similar trend was observed for the native porcine ("25K") amelogenin. Porcine "23K" amelogenin was only sparingly soluble from pH of 4.0 to 9.0, in contrast to the analogous recombinant "rM166", which was more soluble in acidic solutions. The synthetic amelogenin polypeptide "TRAP" was extremely insoluble, while synthetic LRAP was readily soluble. Porcine "20K" amelogenin solubility increased strikingly as the solution pH was lowered from 7.0 to 6.0. Increasing IS decreased the solubility of rM179. While Zn2+ reduced rM179 solubility, Ca2+ and Mg2+ showed no significant effects. We conclude that the solubility of amelogenin was dependent on the primary structure, solution pH, and IS, and the low solubility of amelogenins under physiological conditions may result from their tendency to form quaternary (aggregate) structures in vivo.
The Process of Physical Weakening and Dissolution of the HA-coated Implant in Bone and Soft Tissue
Authors: Ogiso M. , Yamashita Y. , Matsumoto T.
Published: January 2009

Hydroxyapatite (HA)-coated implants were developed to promote osseointegration of titanium implants and to overcome the mechanical drawbacks of solid HA implants. Although many clinical reports on the prognosis of HA-coated implants have reported high success rates, the risks of dissolution and weakening of the coating have been noted. We hypothesized that the chemical and mechanical stability of HA coating are affected by its microstructural characteristics. The present study investigates differences in the microstructures of available HA-coated implants, before and after implantation into the coxal bones of dogs for periods ranging from 3 weeks to 10 months and under the coxal periosteum of dogs for 10 months. The results of transmission electron microscopy and energy-dispersive x-ray analysis revealed that crystallization of super-fine HA crystals occurred in the amorphous phase of the HA coating and progressed over time. This crystallization weakens HA-coated implants by making the amorphous phase brittle, causing stress accumulation within the coating, and causing a decrease in the binding strength between the coating and the substrate. Furthermore, the HA coating dissolved in soft tissue. Dissolution started with the super-fine HA crystals in the crystallized portion that was originally part of the amorphous phase.
Do Dental Composites Always Shrink Toward the Light?
Authors: Versluis A. , Tantbirojn D. , Douglas W.H.
Published: January 2009

Many of the current light-curing composite restorative techniques are rationalized in compliance with the theory that composite shrinks toward the light. Shrinkage directed toward the margins is believed to be responsible for the observed improved marginal properties. However, the dental literature does not consistently support this theory. Experimental determination of contraction patterns is very difficult. In this study, a finite element technique is used to analyze the direction of composite shrinkage as it cures. The process of polymerization can be characterized by pre- and post-gel phases. The stress developed in a restoration can be relieved quickly by the flow of material still in the pre-gel phase. Residual stresses arise after gelation. Both auto- as well as photo-curing composites were analyzed. In photo-curing composites, the gel-point varies throughout the material with the intensity of the light. Experimentally determined light transmittance data for different materials were used in the simulation. Degree of cure and time-dependent shrinkage properties were also included from experimental measurements. The analysis showed that the shrinkage direction was not significantly affected by the orientation of the incoming curing light, but instead was mostly determined by the bonding of the restoration to the tooth and by the free surfaces. Consequently, differences between the contraction patterns of auto- and photo-cure were minimal. It was concluded that composite does not shrink toward the light, but that the direction is predominantly determined by cavity shape and bond quality. Improved marginal properties should be pursued by the optimization of other factors, such as the polymerization process, the curing procedure, and the bond quality. The direction of shrinkage vectors in response to light position does not seem to be an appropriate criterion for the optimization of marginal quality.
Pain Maps from Facial Pain Patients Indicate a Broad Pain Geography
Authors: Türp J.C. , Kowalski C.J. , O'Leary N. , Stohler C.S.
Published: January 2009

Two hundred consecutive female patients, who were referred to a university-based facial pain clinic, were asked to mark all painful sites on sketches showing the contours of a human body in the frontal and rear views. The drawings were analyzed with transparent templates containing 1875 (frontal view) and 1929 (rear view) square cells of equal size. The average patient scored 71.8 cells in the frontal and 99.7 cells in the rear view (corresponding to 3.8% and 5.2% of the maximum possible scores). In individual patient drawings, however, up to 42.7% and 44.9% of all cells were marked. Only 37 cases (18.5%) exhibited pain that was limited to the trigeminal system. An analysis of the pain distribution according to the arrangements of dermatomes revealed three distinct clusters of patients: (1) pain restricted to the region innervated by the trigeminal nerves (n = 37); (2) pain in the trigeminal dermatomes and any combination involving the spinal dermatomes C2, C3, and C4, but no other dermatomes (n = 32); and (3) pain sites involving dermatomes in addition to the ones listed above (n = 131). Mean ages in the three clusters were 38.7, 35.5, and 37.5 years, respectively (p = 0.62, n.s.). Widespread pain existed for longer durations (median, 48 months) than conditions involving local and regional pain (median, 24 months) (p = 0.02, s.). Our findings showed that among a great percentage of persistent facial pain patients the pain distribution is more widespread than commonly assumed, and that the persistence of pain in the regional and widespread pain presentations is significantly greater than in cases with pain limited to the trigeminal system.
The Epithelium-Tooth Interface— A Basal Lamina Rich in Laminin-5 and Lacking Other Known Laminin Isoforms
Authors: Hormia M. , Sahlberg C. , Thesleff I. , Airenne T.
Published: January 2009

The attachment of the marginal gingiva to the tooth surface is mediated by a thin nonkeratinized epithelium termed the junctional epithelium (JE). Ultrastructural studies have revealed that the attachment of the JE to the tooth surface occurs through hemidesmosomes (HD) and a basal lamina-like extracellular matrix termed the internal basal lamina (IBL). We have previously shown that neither type IV collagen nor prototypic laminin, two common components of basement membranes (BM), is present in the IBL between the epithelium and the tooth. In the present study, we show that laminin-5 is a major component of the IBL in both rodent and human tissues. By using in situ hybridization, we also show that the cells of the JE express the LAMC2 gene of laminin-5. In other parts of gingival epithelium, LAMC2 gene expression is less prominent. Our results indicate that the epithelium-tooth interface is a unique structure wherein epithelial cells are induced to secrete a basal lamina containing laminin-5 and no other presently known laminin isoform.
HLA-DR Alleles are Associated with IDDM, but not with Impaired Neutrophil Chemotaxis in IDDM
Authors: Gustke C.J. , Stein S.H. , Hart T.C. , Hoffman W.H. , Hanes P.J. , Russell C.M. , Schuster G.S. , Watson S.C.
Published: January 2009

Insulin-dependent diabetes mellitus (IDDM) is a risk factor for periodontitis. Depressed neutrophil chemotaxis has been demonstrated in IDDM and in early-onset periodontitis (EOP). HLA-DR antigens are associated with both IDDM and periodontitis. This investigation sought to determine an association of HLA-DR3, -DR4, and -DR53 with impaired neutrophil chemotaxis in an IDDM sample. The neutrophil chemotaxis index of 41 diabetics and 27 controls was determined by a modified Boyden chamber method, and certain class II HLA genotypes were determined by polymerase chain-reaction amplification of genomic DNA by means of sequence-specific primers (PCR-SSP). The mean chemotaxis index of the diabetics was significantly less than that of the controls (p < 0.02). HLA-DR3 (p ≤ 0.002), -DR4 (p < 0.003), and -DR53 (p < 0.001) were associated with IDDM. Neutrophil chemotaxis and glucose metabolism were not significantly correlated. None of the HLA-DR alleles was associated with impaired neutrophil chemotaxis. Therefore, the neutrophil chemotaxis defect of IDDM appears to be independent of these HLA-DR-associated genes.