bluuebunny/crossref_metadata_2025 huggingface dataset:
Resin Permeation into Acid-conditioned, Moist, and Dry Dentin: A Paradigm using Water-free Adhesive Primers
Authors: Tay F.R. , Gwinnett A.J. , Pang K.M. , Wei S.H.Y.
Published: January 2009

Preservation of the morphological integrity of demineralized dentin collagen in its hydrated state may account for the success observed in wet-bonding procedures. This study investigated the micromorphological differences between moist- and dry-bonding techniques with the use of: (a) Aelitebond, an alcohol-based, water-free, single-component dentin adhesive primer system; and (b) a water-free, acetone-based experimental primer similar to the acetone-based, water-containing All-Bond 2, a two-component primer system. In the wet groups, acid-conditioned dentin surfaces were blotted so that they remained visibly moist prior to bonding. In the dry groups, dentin surfaces were air-dried for 30 sec. Following the bonding procedures, dentin discs in each group were laminated together by means of a chemical-cure resin and processed for scanning electron microscope (SEM) and transmission electron microscope (TEM) examination. Conditioning with 10% H3PO4 for 20 sec. produced complete demineralization of the outer dentin. In the wet groups, banded collagen and interfibrillar spaces could be observed at the surface of the acid-conditioned dentin. Complete wetting of the loosely arranged collagen fibrils by the resin resulted in the formation of a hybrid layer. In the dry groups, only a very thin hybrid layer was observed on the dentin surface, along the walls of the tubules, and along the course of their lateral branches. The absence of banded collagen and interfibrillar spaces within these areas suggested the existence of a collapsed dentin matrix along various liquid-vapor boundaries that restricted resin permeation into the subsurface intertubular matrix, producing an incompletely infiltrated "hybridoid region".
The Effect of Demineralized Bone Matrix on the Healing of Intramembranous Bone Grafts in Rabbit Skull Defects
Authors: Rabie A.B.M. , Deng Y.M. , Samman N. , Hagg U.
Published: January 2009

A clinical dilemma exists regarding the type of bone that should be used to replace diseased or traumatized osseous tissue. Oral, plastic, and orthopedic surgeons normally implant viable mineralized endochondral (EC) autografts or demineralized EC allografts. A few clinicians have recognized the disadvantages of using EC bone in craniofacial surgery and advocated the replacement of intramembranous (IM) bone with healthy IM bone. However, controversy and uncertainty surround our understanding of these matrices to induce bone formation. Recent studies have advocated the use of other materials with osteoinductive properties, such as demineralized bone matrix (DBM). The proposed delivery system used in this study included IM bone grafts, DBM, and fixation of the IM bone graft. The purpose of this work was to gain further insights into the mechanism of healing of IM bone, in both the presence and the absence of DBM, and to compare the healing of IM bone grafts with that of DBM alone. Critical-sized (10 x 5 mm), full-thickness bony defects in rabbit parietal bone, devoid of periosteum, were filled with IM bone graft (mandible) alone, demineralized cortical bone matrix (DBM) alone, or combined DBM-IM bone graft, or were left unfilled. Histologic changes were examined 14 days later. The IM bone graft healed through IM ossification with no intermediate cartilage stage. DBM and composite DBM-IM healed through an EC ossification with an intermediate cartilage stage. It is hypothesized that the role of the IM graft is to induce neovascularization into the defect site, and that the undifferentiated mesenchymal cells in the perivascular region of the new blood vessels are induced by the bone morphogenetic protein(s) in the DBM into bone-forming cells.
Highly Virulent Strains of Herpes Simplex Virus Fail to Kill Mice Following Infection via Gingival Route
Authors: Monma Y. , Chen Z.J. , Mayama H. , Kamiyama K. , Shimizu F.
Published: January 2009

Virulence ot herpes simplex virus (HSV) in mice has been demonstrated to be dependent on the site of infection. In this experiment, pathogenesis of HSV was studied in 2 different routes of infection in a mouse model system. When BALB/c mice were infected with 5 x 103 plaque-forming units (PFU) of virulent HSV type 1 Miyama GC+ strain (HSV-1-GC+) intraperitoneally, all mice were dead in 6 to 9 days. HSV-1-GC+ was recovered from organs such as the cerebrum, cerebellum, brainstem, and spleen 2 to 5 days after infection, but not from other organs such as trigeminal ganglia. However, if mice were infected in the maxillary gingiva with 1.0 x 107 PFU of HSV-1-GC+, all mice survived. HSV-1-GC+ was recovered from the trigeminal ganglia and brainstem 2 to 5 days after infection, but not from other organs tested. When mice were infected in maxillary gingiva with HSV-1-GC+, followed by the intraperitoneal injection of 6 mg of cyclophosphamide 72 hrs after virus infection, all mice were dead within days. Immunofluorescent and hematoxylin-eosin staining of gingival tissue sections revealed that when mice were infected in maxillary gingiva with HSV-1-GC+, 3 times as many γδ T-cells and 5 times as many polymorphonuclear cells can be detected in sections of maxillary gingiva when compared with non-infected mice. These data show that the gingiva of mice is considerably more resistant to infection with HSV, compared with the peritoneal cavity, and suggest the possible presence of an oral defense mechanism which might be different from that in the peritoneal cavity.
The Flow Rate of Unstimulated Human Labial Gland Saliva
Authors: Ferguson D.B.
Published: January 2009

Although the minor salivary glands contribute an important fraction of the resting flow of saliva, which may be particularly relevant in xerostomia, there is currently no precise method of measuring the flow from individual glands or recording the number of active glands in a given area. These experiments were devised to test a photographic method of assaying labial gland saliva and to see whether subjects of different ages had differing flow rates and/or numbers of glands. Unstimulated saliva secretion from labial salivary glands was measured in healthy non-medicated subjects aged 20 to 55 years by a photographic method. The number of glands in a unit area, the individual gland flow rates, and the flow rates per unit area were assessed. There was a high correlation between flow rate per unit area as measured by this method and measurements from weighing filter paper before and after absorption of the saliva droplets when the latter method could be used. The mean flow rate for 22 subjects was 2.35 ± 1.65 uL/min over the whole area photographed. Subjects with higher flow rates had both more active glands and a higher rate of secretion from individual glands. There was no correlation between either number of active glands in a unit area or of flow rate and age in the age range represented by these subjects.
Gene Defect in Hypodontia: Exclusion of EGF, EGFR, and FGF-3 as Candidate Genes
Authors: Arte S. , Nieminen P. , Pirinen S. , Thesleff I. , Peltonen L.
Published: January 2009

Hypodontia, congenital absence of one or a few permanent teeth without any systemic disorders, is regarded as an autosomally inherited dominant condition with varying expression and incomplete penetrance. Many studies have reported that the prevalence of hypodontia varies from 5% to 10% among European and Asian populations. The teeth most often missing are second premolars, upper lateral incisors, and lower central incisors. Consequently, we call this trait incisor-premolar hypodontia. Peg-shaped or strongly mesio-distally reduced upper lateral incisors demonstrate variation in the expression of the trait. The gene or genes causing incisor-premolar hypodontia are not known. We have begun the genetic mapping of hypodontia by using linkage analyses in seven Finnish three-generation families with 77 individuals, 31 affected with incisor-premolar hypodontia. As the first step, we studied the possibility of linkage between hypodontia and some candidate genes which have been suggested to have important functions during tooth development. Here we report the exclusion of EGF, EGFR, and FGF-3 loci as possible sites for gene mutation causing incisor-premolar hypodontia in our family material. Because of the close location of the FGF-3 and FGF-4 genes, the results also suggest the exclusion of the FGF-4 locus.
Identification of the Fas Antigen in Human Gingiva
Authors: Yoshioka C. , Muraki Y. , Fukuda J. , Haneji T. , Kobayashi N.
Published: January 2009

The Fas antigen is a cell-surface glycoprotein that mediates apoptosis from the cell surface into the cytoplasm. Polyclonal antibody (Fas D) was raised against a synthetic polypeptide selected from the extracellular part of the human Fas antigen (amino acid residues 104-114) and was used to detect the Fas antigen in human gingiva. Biopsy specimens of human gingiva were prepared, and the paraffin sections were reacted with the Fas D antibody by an immunohistochemical method. The antibody localized to the prickle-cell layer and to granular layer keratinocytes of human gingiva. Proteins were also prepared from human gingiva and subjected to SDS-PAGE, followed by Western-blotting analysis with the Fas D antibody. The antibody interacted with a band corresponding to an estimated molecular weight of 35 kDa. The incidence of the immunoreactive 35-kDa protein was detected in the gingiva of 90% of the 20 individuals examined. The Fas antigen detected in human gingiva may be related to the physiological turnover of oral mucosa.
The Threshold Effects of Nd and Ho:YAG Laser-induced Surface Modification on Demineralization of Dentin Surfaces
Authors: Kinney J.H. , Haupt D.L. , Balooch M. , White J.M. , Bell W.L. , Marshall S.J. , Marshall G.W.
Published: January 2009

Laser irradiation alters the structure of dentin and produces surface layers that give the appearance of being more enamel-like. The laser-modified surface may be more resistant to demineralization; hence, many investigators are proposing continued development of the laser as a possible preventive treatment for caries. The purpose of this study was to explore the morphological changes that occur in dentin when treated at threshold illuminance with two clinically interesting laser wavelengths, and to evaluate the effectiveness of the laser-treated surface at resisting demineralization in an acid-gel solution. The Nd:YAG laser (wavelength 1060 nm) produced significant recrystallization and grain growth of the apatite, without the formation of second phases such as β-tricalcium phosphate. This recrystallized surface layer showed resistance to demineralization; however, the layer did not provide protection of the underlying dentin from demineralization because of cracks and macroscopic voids that allowed for penetration of the demineralizing gel. The Ho:YAG laser-treated surface (wavelength 2100 nm) did not show significant evidence of recrystallization and grain growth, and only a trace amount of an acid-resistant layer was observed with demineralization. It is speculated that the Ho:YAG laser is coupling with absorbed water, and that the heat transfer from the water to the mineral phase is inefficient. For the purposes of creating a demineralization-resistant layer, threshold illuminance with both Nd:YAG and Ho:YAG was ineffective.
Dentin Bonding: SEM Comparison of the Resin-Dentin Interface in Primary and Permanent Teeth
Authors: Nor J.E. , Feigal R.J. , Dennison J.B. , Edwards C.A.
Published: January 2009

Previous studies have suggested minor differences between primary and permanent teeth in terms of dentin composition and morphology. Other reports indicated lower bond strengths of resin composites to dentin of primary teeth compared with dentin of permanent teeth; however, no information is available regarding differences in the micromorphology of the resin-dentin interface that may explain these lower bond strengths. Therefore, the purpose of the present study was to compare primary and permanent teeth in terms of the thickness of the hybrid layer developed with two bonding systems. Our hypothesis was that bonding differences previously reported between primary and permanent dentin would be reflected in hybrid layer differences observable in SEM analyses. Twenty human extracted and non-carious teeth were divided into 4 groups: 5 primary and 5 permanent teeth restored with All-Bond 2/Bisfil P system; and 5 primary and 5 permanent teeth restored with Scotchbond Multi-Purpose/ZlOO. The sample area available on each tooth was divided for the two dentin conditioning times (7 and 15 sec). Measurements of hybrid layer thickness were performed by means of SEM at xl3,000. The results of this study indicated that the hybrid layer produced is significantly thicker in primary than in permanent teeth (p = 0.0001), suggesting that primary tooth dentin is more reactive to acid conditioning. No difference was observed in the hybrid layers produced by the two adhesive systems (p = 0.7920). The increased thickness of the hybrid layer in primary teeth (25 to 30%) and the subsequent lack of complete penetration of adhesive resin into previously demineralized dentin may contribute to the lower bond strengths to primary dentin reported in the literature. If a narrower hybrid layer more uniformly infused with resin is the goal of dentin bonding, it is concluded that a differentiated protocol for bonding to primary dentin (with shorter time for dentin conditioning) can be used as a means to reproduce the hybrid layer thickness seen in permanent teeth.
Microcracks in Dental Porcelain and Their Behavior during Multiple Firing
Authors: Mackert J.R. , Williams A.L.
Published: January 2009

Dental porcelains rely on the high-thermal-expansion mineral leucite to elevate their bulk thermal expansion to levels compatible with dental PFM alloys. The microcracks that form around these leucite particles when cooled during porcelain manufacture are a potential source of change in bulk porcelain thermal expansion during fabrication of porcelain-fused-to-metal crowns and bridges. The purpose of the present study was to determine whether multiple firings of commercial dental porcelains could produce changes in microcrack density. Specimens of six commercial porcelains and the "Component No. 1" of the Weinstein patent were fabricated and subjected to 1, 2, 4, 8, and 16 firings. The microcrack densities were determined by quantitative stereology, whereby intersections of microcracks were counted with a test grid. The microcrack data were subjected to linear regression analysis and analysis of variance. The microcrack densities of four of the six porcelains and the Component No. 1 frit were not significantly affected by the number of firings ( p > 0.05). One porcelain exhibited a weak but highly significant positive correlation between microcrack density and multiple firings ( r2 = 0.24, p = 0.0003), while the remaining porcelain exhibited a weak but statistically significant negative correlation between microcrack density and multiple firings ( r2 = 0.15, p = 0.006). The results of this study indicate that even for porcelains that exhibit a measurable change in microcrack density as a function of multiple firings, the magnitude of the increase or decrease in microcrack density after several firings is sufficiently small to cause only negligible shifts in porcelain bulk thermal expansion.
Cell Proliferation and Expression of EGF, TGF-α, and EGF Receptor in the Developing Primary Palate
Authors: Iamaroon A. , Tait B. , Diewert V.M.
Published: January 2009

Growth factors such as epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-a) play an important role in cell proliferation during embryogenesis. The purposes of the study were to characterize the expression patterns of EGF and TGF-a and their receptor, EGF receptor (EGF-R), and to analyze regional patterns of cell proliferation during primary palate morphogenesis when facial primordia outgrow and fuse to form the premaxillary and upper lip regions. The expression of all molecules was studied with indirect immunohistochemistry with conventional and/or confocal microscopes in normal days 10 and 11 CD1 mice. 5-Bromodeoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA) were used as markers of cell proliferation. EGF, TGF-a, and EGF-R were found to have similar distribution patterns at all stages examined. In the anterior region of the face, the molecules were intensely localized at the tips and peripheral regions of the medial and lateral nasal prominences. Upon fusion of the facial prominences, all three molecules were present mainly at the fusion area and the tips and peripheral areas of the maxillary and nasal prominences. BrdU and PCNA were found to have distribution patterns similar to those of EGF, TGF-a, and EGF-R, with intense staining at the tips and peripheral regions of the facial prominences. These results show that EGF, TGF-a, and their receptor were expressed more intensely in regions of the developing primary palate where cell proliferation was most pronounced, and suggest that EGF, TGF-a, and EGF-R may play a role in cell proliferation during morphogenesis of the primary palate.
Cariogenicity of Streptococcus mutans Strains with Defects in Fructan Metabolism Assessed in a Program-fed Specific-pathogen-free Rat Model
Authors: Burne R.A. , Chen Y.-Y.M. , Wexler D.L. , Kuramitsu H. , Bowen W.H.
Published: January 2009

To define the role of dental plaque fructans and the enzymes involved in their metabolism in the initiation and progression of dental caries, we constructed otherwiseisogenic mutants of Streptococcus mutans defective in the ability to synthesize fructans, to degrade fructans, or to do both. The cariogenic potential of these organisms was evaluated in a specific-pathogen-free rat model in which the feeding patterns of the animals were controlled by means of a Konig-Hofer programed feeder. Specifically, rats were infected with wild-type S. mutans UA159 or derivatives of this strain which contained an insertionally-inactivated fructanase ( fruA) gene, fructosyltransferase ( ftf) gene, or which had both genes inactivated. The animals were fed 17 meals per day of Diet 2000 containing 56% sucrose at 70-minute intervals for five weeks, and caries experience was evaluated. Animals infected with S. mutans with a mutated fruA gene only had statistically significant decreases in sulcal caries severity. Such a decrease was not observed in previous studies with ad libitum-fed animals (Wexler et al., 1992). The manifestation of diminished virulence in the programed feeding model, but not in ad libitum-fed animals, supports the concept that the primary contribution of FruA to virulence is through the utilization of fructans storage polysaccharides. Animals infected with strains carrying the ftf mutation or simultaneous mutations in ftf and fruA did not display decreased virulence, perhaps indicating that sucrose utilization pathways may compete for substrate in vivo, or that accumulation of fructans may affect the ecology or the physicochemical characteristics of dental plaque in such a way as to reduce its cariogenic potential. The results of this study also emphasize that the contribution of a particular virulence determinant to the caries process may be highly dependent on the experimental design, feeding regimen and diet, and the presence or absence of other enzymatic activities.
Early and Long-term Wear of Conventional and Resin-modified Glass Ionomers
Authors: de Gee A.J. , van Duinen R.N.B. , Werner A. , Davidson C.L.
Published: January 2009

Various studies have shown that glass ionomers are susceptible to brittle fracture and acid conditions and that they undergo long-term changes in their mechanical properties. Little information is available on how brittleness, acid susceptibility, and long-term changes are reflected in the wear characteristics of glass ionomers. The purpose of this study was to evaluate long-term changes in conventional glass ionomers, metal-reinforced glass ionomers (including a cermet), and (light-curing) resin-modified glass ionomers by wear experiments simulating the wear process in occlusal contact-free areas. The wear tests were conducted periodically over a period of one year. In addition, wear was determined after one year at a pH of 5 or 6, for assessment of acid susceptibility, and at a condition as found in the occlusal contact areas. All materials showed high early-wear rates which decreased significantly during the one-year test period. This long-term process may be related to a slow progression of the acid-base reaction extending over several months. At each stage, the resin-modified glass ionomers wore significantly faster than the acid-base setting glass ionomers. Most of these materials were not affected at a pH of 6.0, while at a pH of 5.0 only the conventional and the metal-reinforced glass ionomers showed increased wear. Direct contacts with the antagonist led to a significant increase in wear in comparison with contact-free wear, probably as a result of sub-surface fatigue phenomena. In view of the unfavorable wear characteristics of the resin-modified glass ionomers and the high early wear of the conventional glass ionomers, including the metal-reinforced glass ionomers, it was concluded that none of these materials can yet be recommended for use in high-stress-bearing situations.
Serendipity or Luck: Stumbling on Gingival Crevicular Fluid
Authors: Krasse BO
Published: January 2009

The word "serendipity" comes from the tale "The Three Princes from Serendip" by Horace Walpole. It is defined as "an aptitude for making fortunate discoveries accidentally". The demonstration of crevicular fluid was truly an accidental discovery. Niels Brill, a successful private practitioner in Copenhagen with prosthodontics as a specialty, and Bo Krasse, associate professor of cariology, were examining the microflora of gingival pockets in a dog. Before the samples were taken, gingiva and teeth were disinfected with an iodine solution. The solution disappeared from different pockets at different speeds. After intravenous injection of a fluorescein solution, the occurrence of fluorescein could be recorded on filter paper strips. Thus, after stumbling on the disappearance of iodine from the cervical areas of the teeth, two nonperiodontologists managed to demonstrate why it disappeared. This article describes the details of the discovery and subsequent studies, and offers some reflections by one of the original authors 37 years later.
Glucose-mediated Alteration of Cellular Function in Human Periodontal Ligament Cells
Authors: Nishimura F. , Terranova V. , Foo H. , Kurihara M. , Kurihara H. , Murayama Y.
Published: January 2009

Because diabetic patients are easily led to manifest severe periodontitis, we wanted to determine whether various glucose levels interfere with normal cellular function. Human periodontal ligament (PDL) cells were cultured in glucose-free medium, or in medium containing either 1100 mg/L of glucose (normal-glucose medium) or 4500 mg/L of glucose (high-glucose medium). Cells cultured in glucose-free medium changed their morphology from spindle-shaped to round, and incorporated trypan blue in a time-dependent manner. The incorporation rate was much faster in cells with shorter cell cycles than in those with longer cycles, suggesting the involvement of cell-cycle progression in cell death. However, fragmented DNA, which suggests apoptotic cell death, was not observed in these cells. We reasoned that initial cell rounding and detachment from the culture plate might be due to the conformational changes in cell-surface receptors to fibronectin, a major extracellular matrix for fibroblasts. Western blot analysis revealed that cells cultured in glucose-free medium lost their fibronectin receptor in a time-dependent manner. In addition, fibronectin receptor expression was much higher in cells cultured in high-glucose medium than in cells cultured in normal-glucose medium. Furthermore, the over-expression of the fibronectin receptor resulted in a suppressed chemotactic response of these cells to platelet-derived growth factor. On the basis of these data, it was hypothesized that a high glucose level induced over-expression of these receptors. This might be the mechanism by which a high glucose level compromises wound healing in diabetic patients and, at least in part, might be the reason diabetic patients are subject to severe periodontal destruction.
The Chemical Composition of Tooth Enamel in Recessive Dystrophic Epidermolysis Bullosa: Significance with Respect to Dental Caries
Authors: Kirkham J. , Robinson C. , Strafford S.M. , Shore R.C. , Bonass W.A. , Brookes S.J. , Wright J.T.
Published: January 2009

Previous reports have linked the prevalence of tooth abnormalities with high caries experience in the different types of epidermolysis bullosa (EB). However, it is not known to what extent the apparent susceptibility to enamel caries is due to disease-related altered enamel chemistry in these cases. The aim of this study was to characterize the enamel of teeth from patients suffering from recessive epidermolysis bullosa dystrophica (rEBD) in terms of its mineral content, carbonate content, protein content, and amino acid composition. The results showed that dental enamel from these patients was essentially normal in terms of its chemistry. It is therefore concluded that the high caries experience in recessive dystrophic epidermolysis bullosa patients is probably related to other factors, such as compromised oral hygiene and prolonged oral clearance due to extensive oral soft tissue damage and a cariogenic diet.
Bonding to Intact Dentin
Authors: Nakabayashi N. , Saimi Y.
Published: January 2009

It has been reported that the presence of a smear layer on dentinal substrates can compromise bonding. Typically, smear layers are removed by acidic agents that selectively extract calcium salts from dentin surfaces to leave a collagen-rich substrate. Acid-conditioned dentin (i.e., demineralized) is then primed and an adhesive agent applied. In the present study, we removed smear layers by "polishing" dentin specimens with a hydroxyapatite paste and ultrasonication. Bonding procedures were carried out by means of an aqueous solution of 20% 2-methacryloyloxyethyl phenyl phosphoric acid (phenyl-P) and 30% 2-hydroxyethyl methacrylate, referred to as 20P-30H, a "self-etching primer". The 20P-30H solution was applied to "intact" dentin (i.e., non-demineralized) for either 30 or 60 s. Control samples received no application (0 s) of the self-etching primer. Mean tensile bond strengths (10 MPa) were similar in both the 30-second- and 60-second-primed groups. The widths of formed hybrid layers varied from 0.3 ± 0.2 pm at 0 s application (control) to 2.1 ± 0.3 μm for the 30-second group and 4.1 ± 0.2 pm for the 60-second group. SEM and TEM observations revealed that the 20P-30H self-etching primer created diffusion channels into "intact", calcium-rich dentin which permitted monomer to infiltrate dentin substrates. Hybrid layers identified under microscopic examination demonstrated resistance to both HCl and NaOCI treatments, suggesting that the hybrid layer was not defective, and that bonding was stable.
Cloning, cDNA Sequence, and Alternative Splicing of Porcine Amelogenin mRNAs
Authors: Hu C.-C. , Bartlett J.D. , Zhang C.H. , Qian Q. , Ryu O.H. , Simmer J.P.
Published: January 2009

In mammals, the organic matrix of developing enamel is dominated by amelogenins. To investigate the expression of proteins secreted into the developing enamel matrix, we have constructed a porcine enamel organ epithelia-specific cDNA library. The amelogenin fraction of the cDNA library was characterized by the cloning of amelogenin-specific polymerase chain-reaction (PCR) amplification products, 5' and 3' rapid amplification of cDNA ends (RACE), and by helper phage rescue of unamplified clones. Clones were characterized that encode porcine amelogenin isoforms 173, 157, 56, 41, and 40 amino acids in length. The structure of the porcine amelogenin gene differs from that of any of those yet described. There are two homologous but distinct exons 1, 2, and 7. One of the two exon 7s can vary in length depending upon the selection of either of two polyadenylation signal/cleavage sites. As a rule, a given exon 1 always pairs with the same exon 2 but can be associated with either exon 7. Despite significant sequence divergence within these exons, no differences are observed in exons 3, 5, and 6. We interpret these findings as evidence of a single amelogenin gene expressed from two promoters; however, the results do not exclude the existence of a second amelogenin gene. The variability generated through the use of alternate promoters and exon 7s primarily affects the non-coding regions of the message. A given amelogenin isoform expressed from the two promoters displays four amino acid differences within the signal peptide, while the secreted proteins are identical. Similarly, the alternative use of exon 7 does not alter the structure of the protein products. The pattern of RNA splicing of amelogenin pre-mRNAs is different for the transcripts expressed from the two promoters. The 173- and the 56-residue amelogenins can be expressed from either promoter, while the 157-residue amelogenin is generated by onlv one of the two promoters.
Abnormal Enamel and Pulp Dimensions in Familial Dysautonomia
Authors: Mass E. , Zilberman U. , Gadoth N.
Published: January 2009

In familial dysautonomia (FD), a hereditary autonomic and sensory neuropathy, somatic growth is impaired. This study was conducted to explore the possibility that tooth dimensions are altered as a consequence of neural crest dysfunction known to be present in FD. Enamel, dentin, pulp, and tooth size measurements of mandibular primary and permanent molars from FD patients were compared with those of healthy controls. It was found that although tooth size in the FD patients was smaller than normal, the enamel was thicker on the occlusal table, while the pulp chamber was smaller and disproportional to tooth size. Our results suggest distorted tooth dimensions rather than a generalized growth arrest as observed in other hereditary syndromes, such as Down's or Crouzon.
Human Granulation-tissue Fibroblasts Show Enhanced Proteoglycan Gene Expression and Altered Response to TGF-β1
Authors: Häkkinen L. , Westermarck J. , Kähäri V.-M. , Larjava H.
Published: January 2009

Granulation-tissue fibroblasts are phenotypically unique cells that play an important role in wound repair and the development of chronic inflammatory lesions in connective tissue. In the present study, we compared proteoglycan, type I, and type III procollagen gene expression by granulation-tissue fibroblasts from wound and chronically inflamed tissues with normal gingival fibroblasts. We also analyzed the effect of TGF-β1 on proteoglycan mRNA levels and macromolecule production by these cells. One granulation-tissue fibroblast strain that was composed exclusively of a-smooth-muscle actin-positive cells (myofibroblasts) expressed strongly elevated basal levels of biglycan, fibromodulin, and versican (the large chondroitin sulphate proteoglycan), as well as type I and III procollagen mRNA. TGF-β1 enhanced more potently the expression of types I and III procollagen, biglycan, and versican mRNA by these cells as compared with normal fibroblasts. Other granulation-tissue fibroblast strains, in which about half of the cells expressed a-smooth-muscle actin, also showed enhanced proteoglycan and types I and III procollagen expression as compared with normal fibroblasts. These results suggest that alterations in matrix composition during inflammation and wound healing are regulated partly by altered phenotypes of the cells that produce the matrix, and partly by altered responses of these cells to TGF-β1.
Incorporation of Caseinoglycomacropeptide and Caseinophosphopeptide into the Salivary Pellicle Inhibits Adherence of Mutans Streptococci
Authors: Schüpbach P. , Neeser J.R. , Golliard M. , Rouvet M. , Guggenheim B.
Published: January 2009

The protective effects of milk and milk products against dental caries have been demonstrated in many animal studies. We have shown that this effect was mediated by micellar casein or caseinopeptide derivatives. A reduction in the Streptococcus sobrinus population in the oral microbiota of animals fed diets supplemented with these milk components was consistently observed. A possible explanation for these findings is that milk components are incorporated into the salivary pellicle, thereby reducing the adherence of S. sobrinus. This hypothesis was tested in vitro by the incubation of bovine enamel discs with unstimulated saliva. The resulting pellicle was washed and incubated with caseinoglycomacropeptide (CGMP) and/or caseinophosphopeptide (CPP) labeled with 17- and 12-nm gold particles. All samples were prepared for electron microscopy by high-pressure freezing followed by freeze-substitution. It was demonstrated by high-resolution scanning electron microscopy with back-scattered electron imaging, as well as by transmission electron microscopy, that both peptides were incorporated into the pellicle in exchange for albumin, confirming previous findings. This protein was identified with a mouse anti-human serum albumin followed by goat anti-mouse IgG labeled with 25-nm gold particles. Incorporation of CGMP and/or CPP into sali\ arv pellicles reduced the adherence of both S. sobrinus and S. mutans significantly. It is suggested that the calcium-and phosphate-rich micellar casein or caseinopeptides are incorporated into the pellicle. The resulting ecological shifts, together with the increased remineralization potential of this biofilm, may explain its modified cariogenic potential.
Saliva Protein Binding to Streptococcal Layers Placed at Different Oral Sites in 48 Persons
Authors: Rudney J.D. , Ji Z. , Larson C.J.
Published: January 2009

Preliminary studies of 10 subjects suggested that saliva protein binding to oral bacteria might vary among oral sites. This study investigated saliva protein binding to layers of oral streptococci in an expanded sample of 48 subjects. Those persons were at opposite extremes for unstimulated whole saliva amylase, sIgA, lactoferrin, and lysozyme in an initial screening of 128 individuals. Layers of Streptococcus gordonii Blackburn or Streptococcus oralis 10557 on enamel chips were placed on buccal left and right upper premolars and molars (UL, UR), labial upper central incisors (UC), and lingual lower central incisors (LL). After a 10-minute exposure to saliva, bacterial extracts were assayed for bound amylase, sIgA, lactoferrin, and lysozyme. Those proteins also were quantified in unstimulated whole saliva collected after chip exposure. Both strains bound significantly more amylase at UL and UR, and significantly less at UC. Blackburn bound more amylase than 10557 at all sites. Significantly less sIgA was bound at UC; strain differences for sIgA were inconsistent across sites. Significantly more lactoferrin and lysozyme were bound at LL. There were no strain differences for lactoferrin; 10557 bound significantly more lysozyme at UL and UR. Subjects at opposite extremes for saliva protein concentrations differed for bound amylase and lactoferrin; those differences were smaller than site and strain differences. Bound protein levels were correlated across sites and strains. Correlations between whole saliva and bound proteins were moderate and were most consistent at LL. These findings suggest that saliva protein effects on oral ecology may vary among oral sites.
Jaw Movement Responses to Electrical Stimulation of Different Parts of the Human Temporalis Muscle
Authors: Zwijnenburg A.J. , Kroon G.W. , Verbeeten B. , Naeije M.
Published: January 2009

Previous EMG studies have provided indications for the differential activation of the human temporalis muscle. However, in these studies the contribution of different parts of the temporalis muscle could not be separated from the contributions of other muscles, since contraction was performed by voluntary activation. Therefore, the aim of this study was to examine functional differences among various parts of the human temporalis muscle by means of recording the incisal point movement response to electrical stimulation of parts of the muscle. Five healthy male subjects participated in this study. Three locations (anterior, middle, and posterior temporalis muscle) were stimulated, by means of monopolar wire electrodes and rectangular pulses. The insertion depths of the stimulation electrodes were determined by means of magnetic resonance images. Stimulation was performed in four jaw positions (resting position, 50% maximum mouth opening, 1 cm to the left, and 1 cm to the right). Movement responses to stimulation of the different muscle parts were recorded with the OKAS-3D jaw movement analysis system. The movement responses were expressed in polar coordinates. The variation in the direction of the jaw movement response was partly explained by the factors 'stimulation location' and 'jaw position' (ANOVA, p < 0.001). When the stimulation location shifted in an antero-posterior direction, the response changed from a vertical-lateral incisal point movement to a lateral-posterior movement with a smaller vertical component. The jaw position during stimulation also influenced the movement response. A functional subdivision of the temporalis muscle into at least three parts is favored.
Chemokine Expression in Human Oral Keratinocyte Cell Lines and Keratinized Mucosa
Authors: Bickel M. , Nöthen S.M. , Freiburghaus K. , Shire D.
Published: January 2009

Chemoattractant cytokines regulate the immune response within the tissue by recruiting neutrophils and macrophages. These so-called chemokines include a large family of peptide molecules encoded by distinct genes. Their expression is controlled by a variety of microbial and host factors. Among host factors, interleukin-1 (IL-1) is thought to be a key regulator of tissue destruction and mediator of the local immune response. To study its influence on chemokine expression, we used a highly sensitive, semi-quantitative method to assess gene expression at the level of mRNA. RNA was extracted from human oral keratinocyte cell lines after treatment with recombinant human IL-1. To test the method further and possibly establish a chemokine mRNA expression pattern, we also extracted RNA from healthy oral keratinized mucosa. Purified RNA was reverse-transcribed and subsequently amplified in a polymerase chain reaction (RT-PCR) by means of specific primer pairs. Amplified sequences were analyzed by agarose gel electrophoresis, visualized by ethidium bromide staining, transferred to nylon membranes, and hybridized to biotinylated oligonucleotide probes. Detection was achieved by streptavidin-conjugated alkaline phosphatase, a chemiluminescent substrate, and autoradiography. Autoradiographs were analyzed by densitometric measurements. IL-1 stimulation resulted in an increase of the chemokine mRNAs encoding interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and GROγ. Macrophage inflammatory protein-1α (MIP-1α) mRNA was not detectable in keratinocytes. In healthy oral mucosa, we found considerable variation between the subjects. Detection of chemokine mRNAs by RT-PCR proved to be sensitive, specific, and fast. It allows for the study of not only cell-line-derived RNA, but also of RNA isolated directly from biopsy material. The latter feature makes this method well-suited for diagnostic purposes.
Accumulation of Fermentable Sugars and Metabolic Acids in Food Particles that Become Entrapped on the Dentition
Authors: Kashket S. , Zhang J. , Van Houte J.
Published: January 2009

Earlier studies (Kashket et al., 1991) showed that particles of high-starch snack foods remained longer on the teeth than those of high-sucrose, low-starch foods. The question arose whether the prolonged presence of food particles enhances cariogenicity. A study was undertaken to measure sugars, starches, and metabolic acids in retained food particles. Subjects consumed portions of different foods, and particles were removed from all bicuspids and first molars at defined times after swallowing. Dry weights, sugars, and short-chain carboxylic acids were determined. High-sucrose foods were cleared rapidly from the teeth, while high-starch foods were retained for up to 20 min. Sucrose, glucose, and fructose persisted in the retained particles. Particles of high-starch foods accumulated maltose and maltotriose, presumably from the breakdown of starch by salivary amylase. At maximum, maltose plus maltotriose constituted 94% of total sugars in particles of potato chips; corresponding values in doughnuts, peanut butter cookies, and salted crackers were 43, 51, and 61%, respectively. Total fermentable sugars in the particles of high-starch foods were similar to those for the high-sucrose confectionery products. Carboxylic acids accumulated within the particles, presumably due to the fermentation of the sugars by entrapped salivary micro-organisms. At maximum (5 to 7 min), acetic, formic, lactic, and propionic acids rose 17-, 30-, 15-, and 1.3-fold, respectively, in dough-nuts, and to smaller degrees in potato chips, salted crackers, and chocolate-caramel-peanut bars. In summary, the study demonstrated the persistence of sugars, the progressive accumulation of starch breakdown products, and the fermentation of the accumulated sugars in retained food particles. The findings support the view that high-starch foods contribute to the development of caries lesions.
IR and NMR Analyses of Hardening and Maturation of Glass-ionomer Cement
Authors: Matsuya S. , Maeda T. , Ohta M.
Published: January 2009

It has been reported that the silicate phase as well as the cross-linking of the polycarboxylic acid by aluminum and calcium ions played an important role in the hardening of glass-ionomer cement. The objective of this study was to investigate the structural change during hardening of the cements by means of infrared (IR) spectroscopy and solid-state nuclear magnetic resonance (NMR) spectroscopy and to confirm the role of the silica phase in the hardening of the cement. For that purpose, we measured the change in compressive strength of an experimental glass-ionomer cement, two commercial glass-ionomer cements, and a polycarboxylate cement and carried out 29Si and 27 Al NMR analyses of the cement samples after the strength measurement. In the IR spectra during hardening, a characteristic band of the silicate network around 1000 cm-1 shifted toward high frequency with time. The spectrum after hardening was similar to that for a hydrated amorphous silica structure. The 27Al NMR analysis showed that Al3+ ion was tetrahedrally coordinated by oxygen in the original glass, but a part of the Al3+ ion was octahedrally coordinated after hardening to form Al polyacrylate gel. The chemical shift of Si in the 29Si NMR spectra also changed during hardening. The variation in the chemical shift reflected the structural change in the silicate network. The initial increase in compressive strength of the cement was mainly caused by polycarboxylate gel formation. However, it was concluded that the reconstruction of the silicate network contributed to the increase in strength with time during the period after the gelation by cross-linking was completed.
Effect of Cubic Leucite Stabilization on the Flexural Strength of Feldspathic Dental Porcelain
Authors: Denry I.L. , Mackert J.R. , Holloway J.A. , Rosenstiel S.F.
Published: January 2009

Previous studies (Mackert and Evans, 1993) have shown that, when feldspathic dental porcelain is cooled, leucite undergoes a transformation from cubic to tetragonal, associated with a 1.2% volume contraction. This contraction leads to the formation of microcracks in and around the crystals and the development of tangential compressive stresses around the crystals. Our aim was to stabilize increasing amounts of the cubic form of leucite in a leucite-reinforced dental porcelain, evaluate its effect on the flexural strength, and characterize its microstructure. The hypothesis was that in the absence of crystallographic transformation, the contraction of the leucite crystals would be lower, thereby limiting the formation of microcracks and minimizing the development of tangential compressive stresses around the leucite particles. We prepared 8 porcelain compositions by mixing increasing amounts of either leucite (KAlSi 2O6) or pollucite (CsAlSi2O6) with Optec HSP porcelain (Jeneric/Pentron Inc., Wallingford, CT). Porcelain disks were made from each composition (n = 10 per group). X-ray diffraction analyses showed that the amount of stabilized leucite increased with the amount of pollucite added. The microstructure of the specimens containing tetragonal leucite was characterized by twinned leucite crystals, whereas no twinning was observed in the specimens containing cubic leucite. The evaluation of crack deflection showed that significantly less deflection occurred in the specimens containing cubic leucite. The mean biaxial flexural strength for the group corresponding to 22.2 wt% added pollucite, fired at 1038°C, was significantly lower than that for the control group. The group corresponding to 22.2 wt% added leucite fired at 1150°C exhibited a mean biaxial flexural strength significantly higher than that of all other groups that were not significantly different from the control group. Overall, the stabilization of cubic leucite reduced the flexural strength and the number of crack deflections in leucite-reinforced porcelain. Apparently, the development of tangential compressive stresses around the leucite crystals when cooled is responsible for a significant amount of strengthening of feldspathic dental porcelain.
The Estimation of Caries Prevalence in Small Areas
Authors: Leroux B.G. , Maynard R.J. , Domoto P. , Zhu C. , Milgrom P.
Published: January 2009

National surveys have been effective for the estimation of caries prevalence in broad regions of the US. However, it is unclear if data from such surveys can be used to estimate prevalences in small areas such as states or counties because of small sample sizes within individual areas. In this study, we applied specialized statistical methods to the estimation of small-area caries measures using data from an oral health survey conducted in the State of Washington. Dental exams to assess caries and the presence of sealants and fluorosis were performed on 2921 third-grade students in 84 public schools selected by a stratified random sample from all 39 counties in the state. Statistical methods for small-area estimation were used to estimate disease and sealant utilization measures for each of the counties. Adjustment was made for covariates measured at the school level, including ethnicity and the proportion of children in the Federally sponsored school lunch program. Substantial variability in disease and sealant utilization between counties was found. The estimated number of decayed and filled surfaces per child was 4.7 (inter-county range, 2.4 to 7.4). The estimated number of surfaces of untreated decay was 1.2 per child overall (range, 0.5 to 3.1). Thirty percent of the children had restorative treatment needs (range, 15 to 54%). The prevalence of sealants on one or more permanent molars was estimated to be 34% (range, 19 to 46%). Overall, only 8% of children showed evidence of fluorosis. The results demonstrate the usefulness of small-area estimation methods for oral health surveys.