bluuebunny/crossref_metadata_2025 huggingface dataset:
Calcium Blocks Fungicidal Activity of Human Salivary Histatin 5 through Disruption of Binding with Candida albicans
Authors: Dong J. , Vylkova S. , Li X.S. , Edgerton M.
Published: January 2009

Salivary histatin 5 (Hst 5) kills the fungal pathogen C. albicans via a mechanism that involves binding and subsequent efflux of cellular ATP. Our aims were to identify inorganic ions found in saliva that influence Hst 5 fungicidal activity. Increasing ionic strength with relevant salivary anions (Cl− and CO3 −) did not reduce Hst 5 binding or uptake by yeast cells, but reduced the Hst-induced efflux of ATP. Extracellular MgCl2 (25 mM) maximally inhibited 30–40% of Hst 5 killing with 40% reduction in ATP efflux, while pre-treatment of cells with only 2 mM CaCl2 inhibited 80–90% of killing, and prevented ATP efflux. Loss of fungicidal activity by the addition of CaCl2 or MgCl2 was a result of inhibition of binding of Hst 5 to C. albicans cells. Calcium is a potent inhibitor of Hst 5 candidacidal activity at physiological concentrations and may be the primary salivary ion responsible for the masking effect of saliva.
Parathyroid Hormone Protects against Periodontitis-associated Bone Loss
Authors: Barros S.P. , Silva M.A.D. , Somerman M.J. , Nociti F.H.
Published: January 2009

Parathyroid hormone (PTH) functions as a major mediator of bone remodeling and as an essential regulator of calcium homeostasis. In addition to the well-established catabolic effects (activation of bone resorption) of PTH, it is now recognized that intermittent PTH administration has anabolic effects (promotion of bone formation). The aim of this study was to investigate whether intermittent administration of PTH in rodents would block the alveolar bone loss observed in rats when a ligature model of periodontitis is used. Morphometric analysis showed that intermittent PTH administration (40 μg/kg) was able to protect the tooth site from periodontitis-induced bone resorption. In addition, there was a significant reduction in the number of inflammatory cells at the marginal gingival area in sections obtained from animals receiving PTH compared with control animals. These findings demonstrated that intermittent PTH administration was able to protect against periodontitis-associated bone loss in a rodent model.
Proteolysis of ICAM-1 on Human Oral Epithelial Cells by Gingipains
Authors: Tada H. , Sugawara S. , Nemoto E. , Imamura T. , Potempa J. , Travis J. , Shimauchi H. , Takada H.
Published: January 2009

Cysteine proteinases (gingipains) from Porphyromonas gingivalis are considered key virulence factors of severe periodontitis and host immune evasion. Since expression of intercellular adhesion molecule-1 (ICAM-1) on gingival epithelium is indispensable in polymorphonuclear leukocyte (PMN) migration at the site of periodontitis, we examined the effects of gingipains on the expression of ICAM-1 on human oral epithelial cell lines (KB and HSC-2) by flow cytometry and Western blotting. We found that three purified forms of gingipains efficiently reduced ICAM-1 expression on the cells in a time- and dose-dependent manner. Gingipains reduced the expression on fixed cells and degraded the ICAM-1 in the cell membranes, indicating that the reduction resulted from direct proteolysis. They then disturbed the ICAM-1-dependent adhesion of PMNs to the cells. These results indicate that gingipains cleave ICAM-1 on oral epithelial cells, consequently disrupting PMN-oral epithelial cell interaction, and are involved in immune evasion by the bacterium in periodontal tissues.
TEGDMA Causes Apoptosis in Primary Human Gingival Fibroblasts
Authors: Janke V. , von Neuhoff N. , Schlegelberger B. , Leyhausen G. , Geurtsen W.
Published: January 2009

Previous in vivo studies have revealed that resins may generate a persistent inflammation of oral tissues and cell death as well. Apoptosis is an important regulated process that results in rapid cell death. This study tested the hypothesis that the comonomer triethyleneglycol-dimethacrylate (TEGDMA) causes apoptosis. The effects of TEGDMA on proliferation and apoptosis in primary oral fibroblasts were analyzed by light microscopy and flow cytometry (FACS; Annexin V-assay). TEGDMA at 5 and 7.5 mM inhibited proliferation after 24 hrs. No increased frequency of apoptosis or necrosis was observed with 1 mM or 2.5 mM TEGDMA after 24 hrs. Apoptosis and Annexin V-positive cells were observed with 5 mM and 7.5 mM TEGDMA by light microscopy after 24 hrs. A dramatic increase in apoptotic cells was detected by FACS after 24 hrs with 7.5 mM TEGDMA. Thus, TEGDMA was cytotoxic and “apoptotic” in a dose- and time-dependent manner.
Effect of TEGDMA/BisGMA Ratio on Stress Development and Viscoelastic Properties of Experimental Two-paste Composites
Authors: Feilzer A.J. , Dauvillier B.S.
Published: January 2009

In this study, we explored the reduction of shrinkage stresses in restored teeth by stimulating viscous flow of adhesive restoratives during curing, by increasing the TEGDMA/BisGMA ratio in the resin of composite restoratives. We studied a series of experimental two-paste composites with different amounts of TEGDMA (30, 50, 70 wt%, respectively) in the resin by mechanical testing, infrared spectroscopy, and dilatometry, to determine how comonomer composition affects the mechanical and chemical properties of the composite during curing. It was found that the polymerization rate of BisGMA-TEGDMA composites is indicative of the viscoelastic behavior during curing: The higher the reactivity, the higher the stiffness and viscosity development. Composites with 50 wt% TEGDMA in the resin displayed the highest maximum polymerization rate. High amounts of TEGDMA in the resin only modestly increased the pre-gel viscous flow (= lowered viscosity) property of composites. Of these composites, high post-gel shrinkage is the decisive factor in high shrinkage stress development.
Detection of Vascular Disease Risk in Women by Panoramic Radiography
Authors: Taguchi A. , Suei Y. , Sanada M. , Higashi Y. , Ohtsuka M. , Nakamoto T. , Tsuda M. , Ohama K. , Tanimoto K.
Published: January 2009

Low bone mineral density and rapid bone loss of the skeleton are associated with mortality risk from vascular diseases in post-menopausal women. Panoramic radiographic measurements are considered as indicators of skeletal bone mineral density or bone turnover. We hypothesize that such measurements may be associated with vascular disease risk in post-menopausal women. Associations of mandibular cortical shape and width on panoramic radiographs with skeletal bone mineral density and risk factors related to vascular diseases were investigated in 87 post-menopausal women. Cortical shape was associated with skeletal bone mineral density, low-density lipoprotein cholesterol, apolipoprotein B, resting heart rate, and endothelial dysfunction. Cortical width was associated with skeletal bone mineral density, low-density lipoprotein cholesterol, and apolipoprotein A1. Dentists may be able to refer women with increased risk of vascular diseases, as well as low bone mineral density, to medical professionals for further examination by panoramic findings.
Aging and Secretory Reserve Capacity of Major Salivary Glands
Authors: Ghezzi E.M. , Ship J.A.
Published: January 2009

A loss of acinar cells occurs with aging, while salivary production remains age-stable in healthy adults. It is hypothesized that a secretory reserve exists to preserve function despite a loss of acinar cells in normal aging. The purpose of this double-blind, placebo-controlled, crossover study was to determine age-related differences in salivary response to an anti-sialogogue (glycopyrrolate). Thirty-six healthy subjects (18 young - 20-38 yrs; 18 older - 60-77 yrs) received 4.0 μg/kg IV glycopyrrolate. Parotid and submandibular/sublingual saliva samples and xerostomia questionnaire responses were collected. Variables calculated for each subject were: times to initial and maximum suppression and xerostomic complaint; time to recovery; and durations of suppression and complaint. Salivary function was more adversely affected in older persons. There were no consistent age-associated questionnaire response differences. These findings suggest that salivary gland output is more adversely affected by an anti-sialogogue in healthy older vs. younger adults, supporting the secretory reserve hypothesis of salivary function.
Proteome Analysis of Oral Pathogens
Authors: Macarthur D.J. , Jacques N.A.
Published: January 2009

The oral environment contains diverse communities of micro-organisms including bacteria, fungi, protozoa, and viruses. Studies of oral ecology have led to an appreciation of the complexity of the interactions that oral micro-organisms have with the host in both health and disease. Despite this, diseases such as dental caries and periodontal diseases are still worldwide human ailments, resulting in a high level of morbidity and an economic burden to society. Proteomics offers a new approach to the understanding of holistic changes occurring as oral micro-organisms adapt to environmental change within their habitats in the mouth.
Pro-inflammatory Cytokines Up-regulate MUC1 Gene Expression in Oral Epithelial Cells
Authors: Li X. , Wang L. , Nunes D.P. , Troxler R.F. , Offner G.D.
Published: January 2009

The membrane-bound mucin MUC1 is expressed ubiquitously on epithelial surfaces and is thought to provide protection from bacterial and chemical injury. The present study was undertaken to determine whether MUC1 was expressed in cultured oral epithelial cells and whether expression is modulated by pro-inflammatory mediators released as part of the host response to infection by oral pathogens. Northern and Western blotting experiments showed that KB cells express MUC1 mRNA and protein. When cells were treated with interleukins (IL-1β, IL-6), tumor necrosis factor-alpha (TNF-α), or interferon-gamma (IFN-γ), or combinations of these, real-time PCR demonstrated that MUC1 mRNA increased 1.4- to 3.2-fold. Interestingly, a significant increase in levels of MUC1 protein was also observed. While no effect was observed when KB cells were incubated with LPS from Porphyromonas gingivalis, infection of KB monolayers with this oral pathogen caused a 2.85-fold increase in MUC1 transcript levels. These results suggest that increased MUC1 synthesis may be a key element in the host response to infection with oral pathogens.
Comparison of Stem-cell-mediated Osteogenesis and Dentinogenesis
Authors: Batouli S. , Miura M. , Brahim J. , Tsutsui T.W. , Fisher L.W. , Gronthos S. , Robey P. Gehron , Shi S.
Published: January 2009

The difference between stem-cell-mediated bone and dentin regeneration is not yet well-understood. Here we use an in vivo stem cell transplantation system to investigate differential regulation mechanisms of bone marrow stromal stem cells (BMSSCs) and dental pulp stem cells (DPSCs). Elevated expression of basic fibroblast growth factor (bFGF) and matrix metalloproteinase 9 (MMP-9, gelatinase B) was found to be associated with the formation of hematopoietic marrow in BMSSC transplants, but not in the connective tissue of DPSC transplants. The expression of dentin sialoprotein (DSP) specifically marked dentin synthesis in DPSC transplants. Moreover, DPSCs were found to be able to generate reparative dentin-like tissue on the surface of human dentin in vivo. This study provided direct evidence to suggest that osteogenesis and dentinogenesis mediated by BMSSCs and DPSCs, respectively, may be regulated by distinct mechanisms, leading to the different organization of the mineralized and non-mineralized tissues.
Relative Levels of mRNA Encoding Enamel Proteins in Enamel Organ Epithelia and Odontoblasts
Authors: Nagano T. , Oida S. , Ando H. , Gomi K. , Arai T. , Fukae M.
Published: January 2009

Amelogenin, enamelin, sheathlin (ameloblastin/ amelin), enamelysin (MMP-20), and KLK4 (EMSP-1) are the major structural proteins and proteinases in developing tooth enamel. Recently, odontoblasts were reported to express amelogenin, the most abundant enamel protein. In this study, we hypothesized that odontoblasts express all enamel proteins and proteases, and we measured their relative mRNA levels in enamel organ epithelia and odontoblasts associated with porcine secretory- and maturation-stage enamel by RT-PCR, using a LightCycler instrument. The results showed that amelogenin mRNA in secretory-stage EOE is 320-fold higher than in odontoblasts beneath secretory-stage enamel, and over 20,000-fold higher than in odontoblasts under maturation-stage enamel. Similar results were obtained for enamelin and sheathlin. Enamelysin mRNA levels were equivalent in these two tissues, while KLK4 mRNA was higher in odontoblasts than in secretory-stage EOE. These results support the conclusion that odontoblasts are involved in the formation of the enamel layer adjacent to enamel-dentin junction.
The Human KROX-26/ZNF22 Gene is Expressed at Sites of Tooth Formation and Maps to the Locus for Permanent Tooth Agenesis (He-Zhao Deficiency)
Authors: Gao Y. , Kobayashi H. , Ganss B.
Published: January 2009

Tooth development is mediated by sequential and reciprocal interactions between dental epithelium and mesenchyme under the molecular control of secreted growth factors and responsive transcription factors. We have previously identified the transcription factor Krox-26 as a potential regulator of tooth formation in mice. The purpose of this study was to investigate a potentially similar role for the human KROX-26 orthologue. We cloned the KROX-26 gene and found its single mRNA transcript (2.4 kb) to be expressed in multiple adult tissues. During fetal development, KROX-26 is expressed in the epithelial component of the developing tooth organ during early bud and cap stages as well as in osteoblasts of craniofacial bone and the developing tongue. The KROX-26 gene was mapped to chromosome 10q11.21, a locus that has been associated with permanent tooth agenesis (He-Zhao deficiency). These results indicate a potential function for KROX-26 in the molecular regulation of tooth formation in humans.
MSX1 Gene is Deleted in Wolf-Hirschhorn Syndrome Patients with Oligodontia
Authors: Nieminen P. , Kotilainen J. , Aalto Y. , Knuutila S. , Pirinen S. , Thesleff I.
Published: January 2009

Abnormalities of the short arm of chromosome 4 cause multiple congenital malformations, including craniofacial, oral, and dental manifestations. A candidate gene for oral defects in this region is MSX1, which is mandatory for normal oral and tooth development. We examined the dentition and the presence of MSX1 in eight Finnish patients with abnormalities of 4p, including seven cases of Wolf-Hirschhorn syndrome. Five of the Wolf-Hirschhorn syndrome patients presented with agenesis of several teeth, suggesting that oligodontia may be a common (even though previously not well-documented) feature in Wolf-Hirschhorn syndrome. In fluorescence in situ hybridization (FISH) analysis, the five patients with oligodontia lacked one copy of MSX1, while the other three had two hybridization signals. One of these presented with the only case of cleft palate among the patients. Our result confirms that haploinsufficiency for MSX1 serves as a mechanism that causes selective tooth agenesis but, alone, is not enough to cause oral clefts.
Mouse Cellular Cementum is Highly Dependent on Growth Hormone Status
Authors: Smid J.R. , Rowland J.E. , Young W.G. , Daley T.J. , Coschigano K.T. , Kopchick J.J. , Waters M.J.
Published: January 2009

Cementum is known to be growth-hormone (GH)-responsive, but to what extent is unclear. This study examines the effects of extremes of GH status on cementogenesis in three lines of genetically modified mice; GH excess (giant), GH antagonist excess (dwarf), and GH receptor-deleted (GHR-KO) (dwarf). Age-matched mandibular molar tissues were processed for light microscope histology. Digital images of sections of first molar teeth were captured for morphometric analysis of lingual root cementum. Cross-sectional area of the cellular cementum was a sensitive guide to GH status, being reduced nearly 10-fold in GHR-KO mice, three-fold in GH antagonist mice, and increased almost two-fold in giant mice (p < 0.001). Cellular cementum length was similarly influenced by GH status, but to a lesser extent. Acellular cementum was generally unaffected. This study reveals cellular cementum to be a highly responsive GH target tissue, which may have therapeutic applications in assisting regeneration of the periodontium.
c-fos Expression in Rat Brain Nuclei Following Incisor Tooth Movement
Authors: Magdalena C.M. , Navarro V.P. , Park D.M. , Stuani M.B.S. , Rocha M.J.A.
Published: January 2009

In the rat experimental model, molar tooth movement induced by Waldo’s method is known to cause a temporally and spatially defined pattern of brain neuronal activation. Since orthodontic correction usually involves the entire dental arch, we used a spring-activated appliance to extend the investigation to incisors, and we included brain regions related to antinociception. Adjustment of the non-activated appliance on incisors resulted in c-fos expression in the dorsal raphe, peri-aqueductal gray matter, and the locus coeruleus, in addition to trigeminal sensory subnuclei and the parabrachial nucleus, where neuronal activation has already been detected in previous studies on molar tooth movement. Appliance activation with a 70- g force resulted in a further increase in Fos-immunoreactive neurons in the trigeminal sensory subnucleus caudalis and in the dorsal raphe. This result suggests that there is a recruitment of neurons related to nociception and to antinociception when tooth movement is increased.
Non-primate Lentiviral Vector Administration in the TMJ
Authors: Kyrkanides S. , Kambylafkas P. , Miller J.H. , Tallents R.H.
Published: January 2009

Gene therapy is emerging as a novel treatment method for the management of temporomandibular joint disorders. The aim of this investigation was to study the effects of lentiviral vectors on the temporomandibular joint. Consequently, we injected into the articular joint space a defective feline immunodeficiency virus capable of infecting dividing as well as terminally differentiated cells with the reporter gene lacZ, the expression of which was studied by means of PCR, X-gal histochemistry, and β-galactosidase immunocytochemistry. Our results showed successful transduction of hard and soft tissues of the temporomandibular joint. Interestingly, a subset of primary sensory neurons of the ipsilateral trigeminal ganglion also stained positive for the reporter gene, presumably following uptake of the lentiviral vector by peripheral nerve fibers and retrograde transport to the nucleus. These findings suggest that lentiviral vectors can potentially serve as a platform for the transfer of anti-nociceptive genes for the management of temporomandibular joint pain.
In situ Effect of Frequent Sucrose Exposure on Enamel Demineralization and on Plaque Composition after APF Application and F Dentifrice Use
Authors: Paes Leme A.F. , Dalcico R. , Tabchoury C.P.M. , Del Bel Cury A.A. , Rosalen P.L. , Cury J.A.
Published: January 2009

Since the effect of the combination of methods of fluoride use on enamel demineralization and on plaque composition is not clearly established, this study examined the effect of the combination of acidulated phosphate fluoride (APF) application and F dentifrice on enamel demineralization and on plaque composition. In this crossover study, 16 volunteers, wearing a palatal appliance containing bovine enamel blocks, were subjected to 4 treatment groups: non-fluoridated dentifrice (PD), FD, APF+PD, and APF+FD. The APF was applied to the enamel before the 14-day experimental period. During the experimental period, test dentifrices were applied 3×/day, and a 20% sucrose solution was applied 4× and 8×/day by being dripped on the blocks. Although APF application was able either to increase F concentration in plaque or to reduce the % of mutans streptococci, its combination with F dentifrice use neither reduced enamel mineral loss nor changed any other measured plaque variable with respect to the FD group alone.
Is Fluoride Concentration in Dentin and Enamel a Good Indicator of Dental Fluorosis?
Authors: Vieira A.P.G.F. , Hancock R. , Limeback H. , Maia R. , Grynpas M.D.
Published: January 2009

Despite some studies correlating dental fluorosis (DF) and fluoride (F) concentration in dental enamel, no information is available about DF and dentin F concentration. Our objective was to determine the correlation between teeth F concentration and DF severity in unerupted human 3rd molars, and the correlation between dentin and enamel F concentrations in the same tooth. Ninety-nine 3rd molars were studied—53 from Fortaleza, Brazil (F water, 0.7 ppm), 22 from Toronto (1.0 ppm), and 24 from Montreal (0.2 ppm). DF severity was evaluated according to the Thylstrup-Fejerskov Index, while F concentration was analyzed by Instrumental Neutron Activation Analysis. DF severity varied between TF0 and TF4, while F concentration ranged between 39 and 550 ppm in enamel and 101 and 860 ppm in dentin. Our results showed correlation between dentin F concentration and DF (rS = 0.316, p = 0.001), but no correlation between enamel F concentration and DF (rS = 0.154, p = 0.133). No correlation was observed between dentin and enamel F concentrations in the same tooth (rS = 0.064, p = 0.536).
Characterization of Dental Epithelial Progenitor Cells Derived from Cervical-loop Epithelium in a Rat Lower Incisor
Authors: Kawano S. , Saito M. , Handa K. , Morotomi T. , Toyono T. , Seta Y. , Nakamura N. , Uchida T. , Toyoshima K. , Ohishi M. , Harada H.
Published: January 2009

Dental epithelial progenitor cells differentiate into various cell types during development of tooth germs. To study this mechanism, we produced immortalized dental epithelial progenitor cells derived from the cervical-loop epithelium of a rat lower incisor. The expression patterns of cytokeratin 14, nerve growth factor receptor p75, amelogenin, Notch2, and alkaline phosphatase were examined by immnohistochemistry in both lower and higher cell densities. The patterns of each were compared in the dental epithelium of rat lower incisors. The results demonstrated that these cells could produce ameloblast lineage cells, stratum intermedium cells, stellate reticulum, and outer enamel epithelium. Furthermore, fibroblast growth factor 10 stimulated proliferation of dental progenitor cells and subsequently increased the number of cells expressing alkaline phosphatase. These results suggest that fibroblast growth factor 10 plays a role in coupling mitogenesis of the cervical-loop cells and the production of stratum intermedium cells in rat incisors.
Bacteremia Following Surgical Dental Extraction with an Emphasis on Anaerobic Strains
Authors: Rajasuo A. , Perkki K. , Nyfors S. , Jousimies-Somer H. , Meurman J.H.
Published: January 2009

Our aim was to investigate bacteremia caused by surgical extraction of partly erupted mandibular third molars. From 16 young adults, bacterial samples were taken from the third-molar pericoronal pocket and post-operatively from the extraction socket, and blood samples were drawn from the ante-cubital vein up to 30 min after surgery. Of the subjects, 88% had detectable bacteremia—50% 1 min after the incision, 44% immediately after extraction. The respective percentages at 10, 15, and 30 min were 44%, 25%, and 13%. Blood cultures contained 31 species (74% anaerobes), with 3.9 ± 2.6 species isolated per subject. Most prevalent were the anaerobes Prevotella, Eubacterium, and Peptostreptococcus sp. and the aerobes viridans-group streptococci and Streptococcus milleri group. Any species found in the blood was also isolated from the mouth, from 93% of the pericoronal pockets and from 43% of the extraction sockets. Surgical dental extraction clearly causes bacteremia of a high frequency and lasting longer than thus far assumed.
Dental Caries, Oral Hygiene, and Oral Clearance in Children with Craniofacial Disorders
Authors: Ahluwalia M. , Brailsford S.R. , Tarelli E. , Gilbert S.C. , Clark D.T. , Barnard K. , Beighton D.
Published: January 2009

The reason that children with cleft palates tend to have a greater prevalence of tooth decay than normal children is unclear. We hypothesized that children with cleft palates would have increased oral clearance times for foods and, consequently, higher levels of caries and caries-associated micro-organisms than control children. Children aged 6–16 yrs, with (n = 81) or without (n = 61) cleft palates, were studied. Children with cleft palates had DMFT and dmft scores greater (p < 0.01) than those of the control group. The number of caries-associated organisms was greater in the saliva of the cleft palate children (all p < 0.001). The oral hygiene, plaque and gingival index scores were greater (p < 0.0001), oral clearance was longer (p < 0.01), and levels of sucrose and starch-derived saccharides higher (p < 0.01) in the cleft palate group. However, salivary concentrations of organic acids were lower in the children with craniofacial disorders, probably reflecting the altered physiology of the more mature dental biofilm. The longer oral clearance times of foods and the consequent generation of fermentable sugars from starches may contribute to the higher caries prevalence observed in children with cleft palates.
On the in vitro Fatigue Behavior of Human Dentin: Effect of Mean Stress
Authors: Nalla R.K. , Kinney J.H. , Marshall S.J. , Ritchie R.O.
Published: January 2009

Human dentin is susceptible to failure under repetitive cyclic-fatigue loading. This investigation seeks to address the paucity of data that reliably quantify this phenomenon. Specifically, the effect of alternating vs. mean stresses, characterized by the stress- or load-ratio R (ratio of minimum-to-maximum stress), was investigated for three R values (−1, 0.1, and 0.5). Dentin was observed to be prone to fatigue failure under cyclic stresses, with susceptibility varying, depending upon the stress level. The “stress-life” ( S/N) data obtained are discussed in the context of constant-life diagrams for fatigue failure. The results provide the first fatigue data for human dentin under tension-compression loading and serve to map out safe and unsafe regimes for failure over a wide range of in vitro fatigue lives (< 103 to > 106 cycles).
Gene Expression Profiling of Mouse Condylar Cartilage during Mastication by Means of Laser Microdissection and cDNA Array
Authors: Watahiki J. , Yamaguchi T. , Irie T. , Nakano H. , Maki K. , Tachikawa T.
Published: January 2009

Little is known about the mechanisms of mandibular condylar growth. In this study, gene expression in the mandibular condylar cartilage of young post-natal mice was monitored by means of a cDNA microarray, real-time PCR, and laser microdissection before and after the initiation of mastication (newborn, 7 days, 21 days, initiation of mastication, and 35 days). Insulin-like growth factor-1 (IGF-I), transforming-growth-factor-beta-2 (TGFbeta2), and aggrecan mRNAs were clearly expressed at 21 days, while the expression of osteopontin mRNAs was most clear at 35 days. Parathyroid-hormone-related protein (PTHrP), Indian-hedgehog (Ihh), and insulin-like growth factor-2 (IGF-2) mRNAs were clearly expressed during lactation (newborn and 7 days). Heat-shock-protein 84 (HSP-84) and heat-shock-protein 86 (HSP-86) were clearly expressed at 35 days. These results revealed that gene expression changed during mandibular condylar cartilage growth, and that, interestingly, these changes coincided with the initiation of mastication.
Induction of Apatite by the Cooperative Effect of Amelogenin and the 32-kDa Enamelin
Authors: Bouropoulos N. , Moradian-Oldak J.
Published: January 2009

Extracellular matrix proteins are considered to play essential roles in controlling the nucleation, growth, and organization of hydroxyapatite crystals during enamel formation. The effects of amelogenin and the 32-kDa enamelin proteins on apatite nucleation were investigated by a steady-state gel diffusion device containing 10% gelatin gels loaded with 0, 0.75%, and 1.5% (w/w) native porcine amelogenins. It was found that the induction time for hydroxyapatite precipitation was strongly increased by the presence of amelogenins, suggesting an inhibitory effect of apatite nucleation. Addition of 18 μg/mL of 32-kDa enamelin to 10% gelatin also caused inhibition of nucleation. Remarkably, addition of 18 and 80 μg/mL of 32-kDa enamelin in gels containing 1.5% amelogenin accelerated the nucleation process in a dose-dependent manner. Our observations strongly suggest that the 32-kDa enamelin and amelogenins cooperate to promote nucleation of apatite crystals and propose a possible novel mechanism of mineral nucleation during enamel biomineralization.
Osmotic Blistering in Enamel Bonded with One-step Self-etch Adhesives
Authors: Tay F.R. , Lai C.N.S. , Chersoni S. , Pashley D.H. , Mak Y.F. , Suppa P. , Prati C. , King N.M.
Published: January 2009

One-step self-etch adhesives behave as permeable membranes after polymerization, permitting water to move through the cured adhesives. We hypothesize that osmotic blistering occurs in bonded enamel when these adhesives are used without composite coupling. Tooth surfaces from extracted human premolars were bonded with 5 one-step self-etch adhesives. They were immersed in distilled water or 4.8 M CaCl2, and examined by stereomicroscopy, field-emission/environmental SEM, and TEM. Water blisters were observed in bonded enamel but not in bonded dentin when specimens were immersed in water. They collapsed when water was subsequently replaced with CaCl2. Blisters were absent from enamel in specimens that were immersed in CaCl2 only. Water trees were identified from adhesive-enamel interfaces. Osmotic blistering in enamel is probably caused by the low water permeability of enamel. This creates an osmotic gradient between the bonded enamel and the external environment, causing water sorption into the interface.
Clinical Assessment of Oral Malodor by the Electronic Nose System
Authors: Tanaka M. , Anguri H. , Nonaka A. , Kataoka K. , Nagata H. , Kita J. , Shizukuishi S.
Published: January 2009

A recently developed electronic nose has not yet been clinically applied to evaluations of oral malodor. This investigation sought to determine whether an electronic nose could clinically assess oral malodor. Twenty-nine healthy adults and 49 patients were assessed by results of an actual organoleptic test, a score representing malodor strength with an electronic nose in “top-note” mode (top-note score), and measurements of volatile sulfur compound (VSC) concentrations. The correlation coefficient between top-note and actual organoleptic scores (r = 0.71) was comparable with the log VSC and actual organoleptic scores (r = 0.63). However, the area under the receiver-operating characteristic plots for top-note score was significantly larger than that for log VSC. In logistic regression analyses with top-note score as a dependent variable, probing depth, tongue coating, and plaque control record each had independent associations. Our findings suggest that the top-note score from an electronic nose examination may be useful for the clinical evaluation of oral malodor.
Expression of Adrenomedullin and its Receptors in Human Salivary Tissue
Authors: Kapas S. , Pahal K. , Cruchley A.T. , Hagi-Pavli E. , Hinson J.P.
Published: January 2009

Adrenomedullin is a multifunctional peptide produced by a wide range of different cells and tissues. This study was designed to investigate whether adrenomedullin is present in human saliva and in salivary glands. It was expected that saliva may contain high concentrations of adrenomedullin, which has antimicrobial activity in vitro, which may have functional implications in the oral cavity. Saliva from the submandibular and parotid glands contained higher concentrations of adrenomedullin than did the circulation, but lower concentrations than in whole saliva. This suggests that oral epithelium may contribute the majority of the adrenomedullin peptide found in saliva. Specific adrenomedullin receptors were found in cell lines from the submandibular (HSG) and parotid (HSY) salivary glands. These findings suggest a paracrine/autocrine role for adrenomedullin in these tissues; however, the concentration of adrenomedullin in saliva was insufficient to suggest a significant antimicrobial action in the healthy oral cavity.
Increase in HIV Receptors/Co-receptors/α-defensins in Inflamed Human Gingiva
Authors: Jotwani R. , Muthukuru M. , Cutler C.W.
Published: January 2009

Transmission of HIV-1 through the oral cavity is considered to be a rare event. To identify factors in resistance/susceptibility to oral HIV-1 infection, we analyzed expression in human gingiva of HIV-1 receptors Langerin, DC-SIGN, MR, and GalCer, HIV-1 co-receptors CCCR5, CXCR4, and anti-microbial protein α-defensin-1. Our results show that healthy gingiva is infiltrated with cells expressing all HIV-1 receptors tested; however, there are very few CCR5+ cells and a complete absence of CXCR4+ cells in the lamina propria. In chronic periodontitis (CP), DC-SIGN, MR, CXCR4, and CCR5 increase, but this was accompanied by a ten-fold increase in α-defensin-1 mRNA. The CCR5+ cells were revealed to be T-cells, macrophages, and dermal dendritic cells. Moreover, epithelial expression of GalCer and CXCR4 together was not apical and showed no trend with underlying inflammation. Thus, low expression of HIV-1 co-receptors in health and high expression of α-defensin during CP may comprise endogenous factors that provide protection from oral HIV-1 infection.
Relationship between Oral Sensitivity and Masticatory Performance
Authors: Engelen L. , van der Bilt A. , Bosman F.
Published: January 2009

The size of a bolus determines how it will be manipulated in the mouth and swallowed. We hypothesized that mucosal sensitivity would be important for masticatory function. The accuracy of solid object size perception, spatial acuity, and food particle size reduction during mastication were measured in 22 healthy adults with/without topical anesthesia of their oral mucosa. Topical anesthesia had no effect on the perception of sphere sizes, but significantly reduced spatial sensitivity. Without anesthesia, there was a correlation between an individual’s ability to perceive the sizes of steel spheres (diameter, 4–9 mm) and the sizes of food particles chewed for 15 cycles and at swallowing. There was no correlation between spatial sensitivity and food particle size. We suggest that the stimuli used to test two-point discrimination stimulates only superficial receptors, which involve light touch and are easily anesthetized, while the spheres might excite more deeply-set receptors. The latter appear to be more important for masticatory performance and swallowing.
Nanocrystalline Tetracalcium Phosphate Cement
Authors: Gbureck U. , Barralet J.E. , Hofmann M.P. , Thulĺ R.
Published: January 2009

Calcium hydroxide cements can lack long-term stability and achieve sustained release by matrix-controlled diffusion of hydroxyl ions. Tetracalcium phosphate (TTCP) hydrolyzes slowly to form calcium hydroxide and a thin insoluble apatite layer that prevents further reaction. In this study, mechanical amorphization was used to create a setting calcium-hydroxide-releasing cement from TTCP. The effect of high-energy ball milling of TTCP on the mechanical properties of the cement was investigated. X-ray diffraction data were used to determine the phase composition of the set cements. An accelerated in vitro test compared pH of water after prolonged boiling of nanocrystalline TTCP cements and a calcium salicylate material. As milling time increased, cement compressive strength and degree of conversion increased. Hydroxyl ion release from the cement was comparable with that from a calcium salicylate material. This new cement system offers the antimicrobial potential of calcium salicylate materials combined with the long-term stability of insoluble apatite cements.
Dental Pulp Fibroblasts Contain Target Cells for Lysophosphatidic Acid
Authors: Gruber R. , Kandler B. , Jindra C. , Watzak G. , Watzek G.
Published: January 2009

Lysophosphatidic acid (LPA) is a locally produced bioactive phospholipid which is involved in tissue repair. The objective of this study was to determine whether dental pulp tissue also responds to the phospholipid. Effects of LPA on proliferation, differentiation, and mitogen-activated protein kinase (MAPK) signaling of dental pulp fibroblasts (DPF) were examined in vitro. We report that DPF express LPA receptors LPA1, LPA2, and LPA3 and respond to the ligand with increased mitogenic activity. Involvement of extracellular signal-regulated kinase, p38 MAPK, and c-Jun NH2-terminal kinase in LPA signaling could be demonstrated by use of specific inhibitors and detection of the phosphorylation status of the kinases. An increased mitogenic activity paralleled a decreased number of alkaline-phosphatase-positive cells and expression levels of dentin sialophosphoprotein and osteocalcin. Together, these results suggest that dental pulp fibroblasts can respond to LPA, a process that may play a role in pulp tissue repair.
Clonal Persistence of Oral Fusobacterium nucleatum in Infancy
Authors: Haraldsson G. , Holbrook W.P. , Könönen E.
Published: January 2009

Once established, early-colonizing bacterial species tend to persist in the mouth. To obtain detailed information on the population dynamics of early-colonizing oral anaerobes, we examined the clonal diversity and persistence of clones among oral Fusobacterium nucleatum populations during the first 2 yrs of life. Consecutive salivary samples from 12 infants, collected at 2, 6, 12, 18, and 24 mos of age, yielded a total of 546 F. nucleatum isolates for clonal typing with arbitrarily primed PCR (AP-PCR). Up to 7 AP-PCR types were simultaneously detected in each sample. In 11 out of the 12 infants examined, AP-PCR types persisted for up to 1 yr. Strain turnover rate was high during the first year of life, but then the occurrence of persistent clones increased. This study indicates a wide genetic diversity within the species and provides evidence for the increasing persistence of F. nucleatum clones in the oral cavity with age.
Enhanced Bone Apposition to a Chemically Modified SLA Titanium Surface
Authors: Buser D. , Broggini N. , Wieland M. , Schenk R.K. , Denzer A.J. , Cochran D.L. , Hoffmann B. , Lussi A. , Steinemann S.G.
Published: January 2009

Increased surface roughness of dental implants has demonstrated greater bone apposition; however, the effect of modifying surface chemistry remains unknown. In the present study, we evaluated bone apposition to a modified sandblasted/acid-etched (modSLA) titanium surface, as compared with a standard SLA surface, during early stages of bone regeneration. Experimental implants were placed in miniature pigs, creating 2 circular bone defects. Test and control implants had the same topography, but differed in surface chemistry. We created the test surface by submerging the implant in an isotonic NaCl solution following acid-etching to avoid contamination with molecules from the atmosphere. Test implants demonstrated a significantly greater mean percentage of bone-implant contact as compared with controls at 2 (49.30 vs. 29.42%; p = 0.017) and 4 wks (81.91 vs. 66.57%; p = 0.011) of healing. At 8 wks, similar results were observed. It is concluded that the modSLA surface promoted enhanced bone apposition during early stages of bone regeneration.
Streptococcus mutans Strains Harboring Collagen-binding Adhesin
Authors: Sato Y. , Okamoto K. , Kagami A. , Yamamoto Y. , Igarashi T. , Kizaki H.
Published: January 2009

A previously unidentified 120-kDa protein was detected in Streptococcus mutans strain Z1 and was involved in the cold-agglutination of the strain. We have identified the gene, designated cnm, as being involved in the agglutination of strain Z1 following random mutagenesis. The amino acid sequence of the deduced Cnm protein exhibited high similarity to those of collagen-binding adhesins from staphylococci and other organisms. To confirm whether the protein is involved in collagen-binding, we cloned a cnm gene fragment, overexpressed it in E.coli, and prepared crude extracts. The extracts containing recombinant protein exhibited binding to immobilized collagen and laminin but not to fibronectin. Compared with the parental strain Z1, the cold-agglutination-negative mutant 05A02 exhibited reduced binding to collagen and laminin but retained that to fibronectin. This gene was detected in some strains of S. mutans. Therefore, the cnm gene encoded a new strain-specific member of the collagen-binding adhesin family.
Monocyte Activation by Porphyromonas gingivalis LPS in Aggressive Periodontitis with the Use of Whole-blood Cultures
Authors: Mahanonda R. , Sa-Ard-Iam N. , Charatkulangkun O. , Promsudthi A. , Schifferle R.E. , Yongvanichit K. , Pichyangkul S.
Published: January 2009

In this study, we re-visited the issue of hyper-responsiveness of monocytes to bacterial lipopolysaccharide (LPS) in aggressive periodontitis patients. We used whole-blood cultures to compare monocyte activation by Porphyromonas gingivalis LPS between Thai subjects with generalized aggressive periodontitis and those without periodontitis. Upon stimulation with P. gingivalis LPS, expression of co-stimulatory molecules on monocytes and expression of CD69 on NK and γδ T-cells were analyzed by flow cytometry, and the production of interleukin-1β and prostaglandin E2 was monitored by ELISA. LPS stimulation resulted in a dose-dependent up-regulation of CD40, CD80, and CD86 on monocytes, and up-regulation of CD69 on NK cells and γδ T-cells in both the periodontitis and non-periodontitis groups. The levels of activation markers and the mediator production after LPS stimulation were quite similar for both groups. In conclusion, we did not observe hyper-responsiveness of monocytes to P. gingivalis LPS challenge in Thai patients with aggressive periodontitis.
Characteristics of the Sarcoplasmic Reticulum Ca2+-dependent ATPase from Masticatory Muscles
Authors: Sánchez G.A. , Takara D. , Toma A.F. , Alonso G.L.
Published: January 2009

We compared the sarcoplasmic reticulum (SR) Ca-ATPase from masseter (M) and medial pterygoid (MP) muscles with that from fast muscles (FM) to examine whether its calcium transport capability and enzymatic activity are different. SR vesicles from FM, M, and MP muscles were obtained according to Champeil et al.(1985) . Assays for characterization of the enzyme properties were performed. The results showed similar optimal conditions for the Ca-ATPase activity and calcium transport in M, MP, and FM. However, the maximal values of calcium transport, Ca-ATPase activity, and Ki for thapsigargin were significantly lower in the masticatory muscles. These findings are likely related to different Ca-ATPase isoforms. Since the local anesthetics used in dentistry inhibit Ca-ATPase and calcium transport in FM, it will be important for the effects of these drugs on the Ca-ATPase of masticatory muscles to be assessed.
Salivary Agglutinin/DMBT1SAG Expression is Up-regulated in the Presence of Salivary Gland Tumors
Authors: Bikker F.J. , van der Wal J.E. , Ligtenberg A.J.M. , Mollenhauer J. , de Blieck-Hogervorst J.M.A. , van der Waal I. , Poustka A. , Nieuw Amerongen A.V.
Published: January 2009

Salivary agglutinin (SAG) is encoded by the gene Deleted in Malignant Brain Tumors 1 ( DMBT1) and represents the salivary variant of DMBT1 (DMBT1SAG). While SAG is a bona fide anti-caries factor, DMBT1 was proposed as a candidate tumor-suppressor for brain, digestive tract, and lung cancer. Though DMBT1SAG is expressed in the salivary glands, its expression in salivary gland tumors is unknown. Here we analyzed DMBT1SAG expression in 20 salivary gland tumors and 14 tumor-flanking tissues by immunohistochemistry. DMBT1SAG in salivary gland tumors resembles the changes of expression levels known from DMBT1 in tumors in other cancer types. Particularly, DMBT1SAG was up-regulated in 10/14 tumor-flanking tissues, and a strong staining of the luminal content in the tumor and/or the tumor-flanking tissue was observed in 14/20 cases. This suggests that, in addition to its role in caries defense, SAG may serve as a potential tumor indicator and/or tumor suppressor in salivary gland tissue.
Cortical Neuromagnetic Fields Preceding Voluntary Jaw Movements
Authors: Shibukawa Y. , Shintani M. , Kumai T. , Suzuki T. , Nakamura Y.
Published: January 2009

Slow cortical potentials (readiness potentials, RPs) reflecting the central programming of voluntary jaw movements were reported to appear preceding the movements. However, the current source producing the RP has not yet been localized. This study aimed to determine the cortical regions involved in the central programming of bilaterally symmetrical voluntary jaw movements, by locating the current source of the neuromagnetic counterpart of the RP (readiness field, RF). The RFs were found in the fronto-lateral region bilaterally, starting around 860 and 600 ms prior to the onset of masseter and digastric electromyograms (EMGs), respectively, and gradually increasing in magnitude to the peak within 100 ms before the EMG onset. Thus, the RFs appeared long before the reported onset of the excitability increase of pyramidal tract neurons. The current sources producing the RFs were located in the precentral gyrus bilaterally, with no bilateral differences in strength. We conclude that the primary motor cortex is involved bilaterally in central programming as well as in execution of bilaterally symmetrical voluntary jaw movements.
Amelogenesis Imperfecta in a New Animal Model—a Mutation in Chromosome 5 (human 4q21)
Authors: Seedorf H. , Springer I.N. , Grundner-Culemann E. , Albers H.-K. , Reis A. , Fuchs H. , Hrabe de Angelis M. , Açil Y.
Published: January 2009

Candidate genes for amelogenesis imperfecta (AI) and dentinogenesis imperfecta (DI) are located on 4q21 in humans. We tested our hypothesis that mutations in the portion of mouse chromosome 5 corresponding to human chromosome 4q21 would cause enamel and dentin abnormalities. Male C3H mice were injected with ethylnitrosourea (ENU). Within a dominant ENU mutagenesis screen, a mouse mutant was isolated with an abnormal tooth enamel (ATE) phenotype. The structure and ultrastructure of teeth were studied. The mutation was located on mouse chromosome 5 in an interval of 9 cM between markers D5Mit18 and D5Mit10. Homozygotic mutants showed total enamel aplasia with exposed dentinal tubules, while heterozygotic mutants showed a significant reduction in enamel width. Dentin of mutant mice showed a reduced content of mature collagen cross-links. We were able to demonstrate that a mutation on chromosome 5 corresponding to human chromosome 4q21 can cause amelogenesis imperfecta and changes in dentin composition.
Induction of Calcification in MC3T3-E1 Cells by Inorganic Polyphosphate
Authors: Kawazoe Y. , Shiba T. , Nakamura R. , Mizuno A. , Tsutsumi K. , Uematsu T. , Yamaoka M. , Shindoh M. , Kohgo T.
Published: January 2009

Relatively large amounts of inorganic polyphosphate [poly(P)] (400 μM) have been found in normal osteoblasts. The effect of poly(P) with an average chain length of 65 phosphate residues on cell calcification was therefore investigated with the use of MC3T3-E1 cells. Expression of both osteopontin and osteocalcin was induced by poly(P) (0.1 ~ 1 mM), and cells treated with poly(P) were strongly stained by alizarin red. In addition, the level of alkaline phosphatase activity induced in poly(P)-treated cells was two-fold higher than that in either orthophosphate-treated or control cells but not higher than that in cells treated with β-glycerophosphate and ascorbic acid. In contrast, however, polyphosphatase activities were activated by poly(P) treatment to levels up to six-fold greater than that in controls. MC3T3-E1 cells may utilize poly(P) as a phosphate source for calcification rather than phosphate sources that are mainly produced by ALPase. Poly(P)-dependent induction of polyphosphatase activities may therefore promote calcification in MC3T3-E1 cells.