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Biomedical subjects

T Aoba

Publications and source records attributed to T Aoba.

At least 19 recordsLinked to original sources
PubMed2004

Solubility properties of human tooth mineral and pathogenesis of dental caries.

Dental research over the last century has advanced our understanding of the etiology and pathogenesis of caries lesions. Increasing knowledge of the dynamic demineralization/remineralization processes has led to the current consensus that bacteria-mediated tooth destruction can be arrested or even to some degree reversed by adopting fluoride and other preventive measures without using restorative materials. Our experimental approach provided new insight into the stoichiometries and solubility properties of human enamel and dentin mineral. The determination of the solubility product constant on the basis of the stoichiometric model (Ca)5.x(Mg)q(Na)u(HPO4)v(CO3)w(PO4)3.y(OH,F)1.z, verifies the difference in their solubility properties, supporting the phase transformation between tooth mineral and calcium phosphates in a wide range of fluid compositions as found in the oral environment. Further refinement of the stoichiometry and solubility parameters is essential to assess quantitatively the driving force for de- and remineralization of enamel and dentin in the oral fluid environment. Prediction of the effects of a combination of inhibitors and accelerator(s) on remineralization kinetics is also required. In order to develop devices efficient for optimizing remineralization in the lesion body, it is a critical question how, and to what extent, fluoride can compensate for the activity of any inhibitors in the mineralizing media.

Dental Caries↗
PubMed2002

Dental fluorosis: chemistry and biology.

This review aims at discussing the pathogenesis of enamel fluorosis in relation to a putative linkage among ameloblastic activities, secreted enamel matrix proteins and multiple proteases, growing enamel crystals, and fluid composition, including calcium and fluoride ions. Fluoride is the most important caries-preventive agent in dentistry. In the last two decades, increasing fluoride exposure in various forms and vehicles is most likely the explanation for an increase in the prevalence of mild-to-moderate forms of dental fluorosis in many communities, not the least in those in which controlled water fluoridation has been established. The effects of fluoride on enamel formation causing dental fluorosis in man are cumulative, rather than requiring a specific threshold dose, depending on the total fluoride intake from all sources and the duration of fluoride exposure. Enamel mineralization is highly sensitive to free fluoride ions, which uniquely promote the hydrolysis of acidic precursors such as octacalcium phosphate and precipitation of fluoridated apatite crystals. Once fluoride is incorporated into enamel crystals, the ion likely affects the subsequent mineralization process by reducing the solubility of the mineral and thereby modulating the ionic composition in the fluid surrounding the mineral. In the light of evidence obtained in human and animal studies, it is now most likely that enamel hypomineralization in fluorotic teeth is due predominantly to the aberrant effects of excess fluoride on the rates at which matrix proteins break down and/or the rates at which the by-products from this degradation are withdrawn from the maturing enamel. Any interference with enamel matrix removal could yield retarding effects on the accompanying crystal growth through the maturation stages, resulting in different magnitudes of enamel porosity at the time of tooth eruption. Currently, there is no direct proof that fluoride at micromolar levels affects proliferation and differentiation of enamel organ cells. Fluoride does not seem to affect the production and secretion of enamel matrix proteins and proteases within the dose range causing dental fluorosis in man. Most likely, the fluoride uptake interferes, indirectly, with the protease activities by decreasing free Ca(2+) concentration in the mineralizing milieu. The Ca(2+)-mediated regulation of protease activities is consistent with the in situ observations that (a) enzymatic cleavages of the amelogenins take place only at slow rates through the secretory phase with the limited calcium transport and that, (b) under normal amelogenesis, the amelogenin degradation appears to be accelerated during the transitional and early maturation stages with the increased calcium transport. Since the predominant cariostatic effect of fluoride is not due to its uptake by the enamel during tooth development, it is possible to obtain extensive caries reduction without a concomitant risk of dental fluorosis. Further efforts and research are needed to settle the currently uncertain issues, e.g., the incidence, prevalence, and causes of dental or skeletal fluorosis in relation to all sources of fluoride and the appropriate dose levels and timing of fluoride exposure for prevention and control of dental fluorosis and caries.

Amelogenesis↗
PubMed2001

Sexual dimorphism of porcine amelogenins: male-specific amelogenins have strong adsorption properties onto apatite crystals.

The present studies were undertaken to investigate the sexual dimorphism of porcine amelogenins and to gain information as to whether excesses of male amelogenins, if any, possess functional significance in protein-crystal interactions. Enamel proteins, including the intact full-length amelogenins and their degraded polypeptides, were isolated from the secretory enamel of male and female pigs. To identify the amelogenins among the separated pools of male- and female-matrix proteins, rabbit anti-C13 and C25 peptide sera were used, which reacted specifically with the conserved C-terminal domain. Immunoblotting showed that a few extra members of the amelogenins, sharing common epitopes at the C-terminus, were recognized in male products. The apparent yield of the male amelogenins was only marginal, on the basis of their stained intensities on the gel, but the secreted male amelogenins demonstrated selective (probably the strongest among the amelogenins) adsorption properties onto apatite crystals. Reflecting the general symmetric electrophoretic profiles of the male- and female-enamel proteins in toto, there were no sex-linked differences in the protein-crystal interaction and the resulting regulatory function of crystal precipitation.

Journal Article↗
PubMed2001

The secretion of amelogenins is associated with the induction of enamel and dentinoid in an ameloblastic fibro-odontoma.

Ameloblastic fibro-odontoma is the unique entity of epithelial-ectomesenchymal odontogenic tumors, which is characterized by enamel formation in addition to dentine. We examined immunohistochemically a case of this tumor in which enamel having prism structures was developed in the absence of odontoblast differentiation but was in contact with mesenchymal matrices. Histological examination showed diverse morphological features of epithelial tumor cells, e.g., cuboidal cells comprising tooth bud-like projections, ameloblast- and stellate reticulum-like cells, and residual cells in forms of extended cords or islands of odontogenic epithelium. Immunostaining with anti-amelogenin sera proved that the intracellular production of amelogenins was initiated at the tooth bud-like stage. The secreted amelogenins were detected almost exclusively in the induced enamel and dentinoid areas, as well as in the core region of cementicle-like spheres deposited in the encapsulating stroma. The results obtained indicate that the odontogenic tumor epithelia and its products, i.e., amelogenins, participate in multifaceted aspects of dental hard tissue formation that takes place during oncogenesis.

Adult↗
PubMed2001

Histopathological studies on virulence of dipeptidyl aminopeptidase IV (DPPIV) of Porphyromonas gingivalis in a mouse abscess model: use of a DPPIV-deficient mutant.

To elucidate the role of dipeptidyl aminopeptidase IV (DPPIV) in the virulence of Porphyromonas gingivalis, mice were infected with either a wild-type strain or a DPPIV-deficient mutant using an abscess model. Histopathological analysis of the resulting lesions indicated that DPPIV participates in virulence through the destruction of connective tissue and the less effective mobilization of inflammatory cells.

Abscess↗
PubMed1998

Small-angle X-ray scattering and computer-aided molecular modeling studies of 20 kDa fragment of porcine amelogenin: does amelogenin adopt an elongated bundle structure?

Amelogenins, which are major matrix constituents in the developing tooth, play a regulatory role in the process of enamel crystal formation. Porcine amelogenin with 173 amino acid residues is rich in proline, glutamine, leucine, and histidine. We utilized the small-angle X-ray scattering (SAXS) technique to examine the solution structure of porcine amelogenin. Samples used were two porcine amelogenins with apparent molecular weights of 20 kDa (amino acids 1 to 148) and 13 kDa (amino acids 46 to 148) on SDS-PAGE. Prior to SAXS measurements, the protein samples were dissolved in 2% (v/v) acetic acid to give a concentration range up to 10 mg/ml. Comparison between Rg (the overall radius of gyration) and Rc (the cross-sectional radius of gyration) revealed that the 20 kDa amelogenin exists in this solution as asymmetric particles with a length of about 15 nm, presumably corresponding of dimers. Based on these experimental data and computer-aided molecular modeling studies, we propose that the 20 kDa amelogenin adopts an elongated bundle structure which mainly consists of extended structures similar to polyproline II and/or beta-strand, interspersed with beta-turn or loop.

Amelogenin↗
PubMed1998

Enamel mineralization and an initial crystalline phase.

In this communication, we summarized our recent experimental approaches to an unsettled issue, i.e., the nature and role of an acidic precursor in enamel mineralization. The objectives we specially focused our attention on are: the composition, structure and high resolution images of enamel crystals at various developmental stages, thermodynamic and kinetic consideration of octacalcium phosphate (OCP) vs hydroxyapatite (HA) precipitation in physiological media simulating the enamel fluid, reversible changes in the composition and structure of OCP, effects of fluoride at low concentrations and enamel proteins on OCP hydrolysis, and adsorption of enamel proteins onto OCP and fluoridated hydrolysates at neutral pH and room temperature. On the basis of all experimental evidence, we propose that enamel crystal growth comprises two events: the two-dimensional growth of an OCP-like precursor in a narrow outermost zone adjacent to the ameloblasts and the subsequent overgrowth of apatite units on the template under discrete fluid environment in the underlying region distant from the cell layer. The experimental data also support the concept that the whole process of enamel mineralization is modulated substantially through interaction between enamel proteins and crystals including the acidic precursor.

Adsorption↗
PubMed1997

The effect of fluoride on apatite structure and growth.

Fluoride participates in many aspects of calcium phosphate formation in vivo and has enormous effects on the process and on the nature and properties of formed mineral. The most well-documented effect of fluoride is that this ion substitutes for a column hydroxyl in the apatite structure, giving rise to a reduction of crystal volume and a concomitant increase in structural stability. In the process of enamel mineralization during amelogenesis (a unique model for the cell-mediated formation of well-crystallized carbonatoapatite), free fluoride ions in the fluid phase are supposed to accelerate the hydrolysis of acidic precursor(s) and increase the driving force for the growth of apatitic mineral. Once fluoride is incorporated into the enamel mineral, the ion likely affects the subsequent mineralization process by reducing the solubility of the mineral and thereby modulating the ionic composition in the fluid surrounding the mineral, and enhancing the matrix protein-mineral interaction. But excess fluoride leads to anomalous enamel formation by retarding tissue maturation. It is worth noting that enameloid/enamel minerals found in vertebrate teeth have a wide range of CO3 and fluoride substitutions. In the evolutionary process from elasmobranch through enameloid to mammalian enamel, the biosystems appear to develop regulatory functions for limiting the fluoridation of the formed mineral, but this development is accompanied by an increase of carbonate substitution or defects in the mineral. In research on the cariostatic effect of fluoride, considerable emphasis is placed on the roles of free fluoride ions (i.e., preventing the dissolution and accelerating the kinetics of remineralization) in the oral fluid bathing tooth mineral. Fluoride also has been used for the treatment of osteoporosis, but much still remains to be learned about maximizing the benefit and minimizing the risk of fluoride when used as a public health measure.

Amelogenesis↗
PubMed1996

Recent observations on enamel crystal formation during mammalian amelogenesis.

BACKGROUND: Enamel mineralization taking place during amelogenesis is a unique model to investigate carbonatoapatite formation in vivo. The abundance of proteinaceous crystal growth inhibitors, in particular amelogenins, contributes significantly to the mineralization process. Their putative roles are to prevent random proliferation of crystal nuclei and to regulate the growth kinetics and orientation of the formed enamel crystals. METHODS: The enamel fluid surrounding the forming enamel crystals contains high concentrations of carbonate and magnesium ions, both of which seem to modulate the mineralization process. Particularly, Mg ions can adsorb onto enamel crystal surfaces in a manner to compete with Ca ions. Enamel mineral formed during amelogenesis is featured as calcium-deficient, acid phosphate-rich carbonatoapatites. Currently the most putative stoichiometry model for enamel mineral is (Ca)5-x(HPO4)v(CO3)w(PO4)3-x (OH)1-x. RESULTS: Very significant changes in the morphology, stoichiometry, and solubility of enamel crystals occur during the various stages of amelogenesis. The early enamel mineralization comprises two events: the initial precipitation of the well-documented thin ribbons and the subsequent overgrowth of apatite crystals on those templates. The thin ribbons precipitated in the vicinity of the secretory ameloblasts have the highest contents of acid phosphate, particularly in the form of exchangeable species, whereas both the exchangeable and unexchangeable acid phosphate decrease concomitantly with the progress of the apatite overgrowth and the appearance of elongated hexagonal crystals in the late secretory stages. CONCLUSIONS: Those morphological and compositional features seem to be consistent with the formation of precursors, such as octacalcium phosphate.

Amelogenesis↗
PubMed1996

Disturbed enamel mineralization in a rat incisor model.

Possession of full-thickness hard enamel appears to be one of the indispensable life-saving characteristics of rats. Previous studies by Suga and his colleagues and by others demonstrated that various types of malformation are evoked in continuously erupting rat incisors. In the current report, we directed our effort to oversee various types of enamel malformation caused experimentally in rat incisors. We surveyed the specimens collected by Suga and his colleagues, as well as specimens we obtained. From the results, it is conceivable that perturbation of the programmed sequential events during enamel development is a major factor in the establishment of enamel malformation. Animal studies with either 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) or a multidentate phosphonic acid (EDTPO) confirmed that dentin mineralization provides a certain inductive effect on the secretion of enamel matrix and subsequent enamel crystallization. Our recent studies using anti-microtubular agents led to the conclusion that the acceleration of mineralization in outer enamel is a type of enamel malformation, most likely due to disruption of the cellular regulation of calcium transport under severe toxic regimens. In future work, experimental approaches combining measurements of kinetic factors with static observation of enamel lesions are required before we can gain a comprehensive understanding of the pathogenesis of disturbed enamel mineralization. The kinetic factors to be considered include the rates of tissue apposition and tooth eruption which determine the total volume of tooth substance formed, and the rate of mineral accretion. Furthermore, information as to the composition, crystallinity, solubility, and mechanical properties of enamel defects is needed before we can assess the susceptibility of teeth having those lesions to caries and other physico-chemical attacks in the oral environment.

Ameloblasts↗
PubMed1995

Computer-assisted reconstruction of enamel fissures and carious lesions of human premolars.

The origin of cariogenesis in occlusal fissures remains elusive because of limited information about both the fissure structure and the morphologic details of carious lesions occurring there. The present study was aimed at reconstructing the coronal structure of human premolars with the aid of computer-assisted image analysis and, on the basis of the reconstructed structures, investigating the configuration of fissures, their proximity to the enamel-dentin junction (EDJ), and the occurrence and extension of carious lesions around the fissure walls. The coronal portions of the teeth were embedded in polyester resin and then ground off stepwise by hand. Serial images of the enamel contour on successive ground planes were recorded by objective microscopy after it was distinctly demarcated with either 0.5 w/v% carbol fuchsin or 1 w/v% basic fuchsin. Usually, from 150 to 250 photoprints were collected from a single enamel crown to reproduce the details of the fissure structures. The enamel outline (the occlusal and lateral surfaces, fissure walls, and EDJ) and, if present, the outline of caries-attacked areas were traced accurately on those photoprints by means of a digitizing system. The enamel contour data in series were input into an image analyzing system (Nikon COSMOZONE, 2SA). Observation of the computer-drawn coronal enamel allowed us to view the morphologic features of the fissures (shape, divarication, and proximity to EDJ) and to scrutinize the pathway of enamel caries along fissure walls.(ABSTRACT TRUNCATED AT 250 WORDS)

Dental Enamel↗
PubMed1995

Reversible structural changes of octacalcium phosphate and labile acid phosphate.

Acid phosphate is one of the major impurities incorporated into bioapatites, and its quantity and environment in forming mineral have been used as diagnostic probes to pursue acidic precursor(s). Currently, little is known about the structural feature of nonstoichiometric octacalcium phosphate (OCP), which has been advocated to be, most plausibly, mineral salt initially formed during amelogenesis. In the present report, we attempt to define the state of acid phosphate in OCP crystals which were Ca-deficient and contained 40% total phosphate as acid phosphate. We assessed fractions of acid phosphate in discrete environments by extracting the crystals in either deionized water, 10 mmol/L NaOH solution (initial pH 11), or 150 mmol/L Tris buffer at pH 7.4. Solid samples before and after the treatments were examined by chemical analyses and x-ray diffraction. The results indicated that successive extractions with use of the alkaline solution brought about a reversible change (not hydrolysis) in the interior structure of OCP, which accompanied a marked decrease in acid phosphate. A substantial part of the lost acid phosphate was restored during subsequent treatments at neutral pH, and, intriguingly, this restoration accompanied a re-ordering of OCP structure. The data suggested that the acid phosphate in OCP is separated into three pools: (a) a stable pool corresponding to roughly 50 to 60% of the total acid phosphate, (b) a reversibly exchangeable pool corresponding to 25 to 30% of the acid phosphate which may exist either in the water layer or on crystal surfaces, and (c) an unstable (or irreversibly lost) pool corresponding to 15 to 20% of the acid phosphate, a part of which might be explained by the presence of excess hydrogen in OCP. The present work supports the concept that protons and, to a lesser magnitude, phosphate species can diffuse into and out of the OCP lattice prior to initiation of its hydrolytic transition into apatite.

Apatites↗
PubMed1995

Mechanistic understanding of enamel mineralization under fluoride regime.

In order to learn more about how the microenvironment for enamel mineralization is modified by fluoride at low concentrations (0 through 1 ppm) and how excess fluoride retards the degradation and removal of amelogenins, we studied precipitation reactions in an in vitro model utilizing a dialysis chamber. The results showed that, with the limited supply of Ca ions through the ultrafiltration membrane, the solution composition surrounding the seed crystals showed a proximity to the steady-state condition after 12-24 h equilibration. Major findings were that (a) fluoride overcame partially the inhibition of precipitation and growth reactions by enamel proteins and (b), with this accelerating effect of fluoride, the steady-state Ca concentrations in the media surrounding the seed crystals decreased substantially as a function of fluoride concentration. The overall results support the concept that the presence of fluoride in the mineralizing milieu can modify markedly the steady-state concentrations of mineral lattice ions, particularly decreasing free Ca2+ concentrations, which in turn may modulate protease activities in situ.

Amelogenin↗
PubMed1995

Confocal laser scanning microscopic studies on alveolar bone remodeling with orthodontic tooth movement and retention.

Alveolar bone reconstruction in growing dog during the retention period following orthodontic tooth movement was studied. Three beagle dogs (8-10 kg body weight, about one-year-old) were used and two of the animals were subjected to histological observation. The upper 2nd and lower 3rd premolars on both sides were extracted prior to the orthodontic treatments. After a healing period of one month, the upper 3rd premolar and the lower 4th premolar on the right side were moved mesially with a conventional orthodontic force for 8 weeks, and then retained in their new position for 4 weeks. The contralateral corresponding premolars were used as control. The alveolar bone was double-labeled with tetracycline (TC) during the movement and calcein (Cal) during the retention period. Alveolar bone structure and labeling patterns were examined by contact microradiography, conventional fluorescence microscopy, and confocal laser scanning microscopy (CLSM). Optimizing the separation of TC and Cal labelings in the alveolar bone was attained by the simultaneous use of ultraviolet (364 nm) and argon (488 nm) laser sources for excitation of TC and Cal, respectively. Cal labeling, indicative of new bone deposition showed two distinct patterns: lamination at the periodontal surface and rings circumscribing the vascular canal. The cementum surface also exhibited active deposition during the experimental period. Bone formation was affected by slight changes in magnitude and direction of orthodontic or occlusal forces. CLSM is valuable in deciphering the process of alveolar bone remodeling.

Alveolar Process↗
PubMed1994

Isolation and characterization of a mouse amelogenin expressed in Escherichia coli.

A mouse cDNA encoding a 180 amino acid amelogenin was subcloned into the pET expression plasmid (Novagen, Madison, WI) for production in Escherichia coli. A simple growth and purification protocol yields 20-50 mg of 95-99% pure recombinant amelogenin from a 4.5-liter culture. This is the first heterologous expression of an enamel protein. The expressed protein was characterized by partial Edman sequencing, amino acid composition analysis, SDS-PAGE, Western blotting, laser desorption mass spectrometry, and hydroxyapatite binding. The recombinant amelogenin is 179 amino acids in length, has a molecular weight of 20,162 daltons, and hydroxyapatite binding properties similar to the porcine 173 residue amelogenin. Solubility analyses showed that the bacterially expressed protein is only sparingly soluble in the pH range of 6.4-8.0 or in solutions 20% saturated with ammonium sulfate. The purified protein was used to generate rabbit polyclonal anti-amelogenin antibodies which show specific reaction to amelogenins in both Western blot analyses of enamel extracts and in immunostaining of developing mouse molars.

Amelogenin↗
PubMed1994

Strategies for improving the assessment of dental fluorosis: focus on chemical and biochemical aspects.

In order to assess fluoride accumulation and effects in developing dental tissues, one must determine the concentration profile of fluoride in the tissue and to assess separately the labile (i.e., free ions in fluid and ions associated with organic matter) and stable (i.e., incorporated into apatite lattice) pools of fluoride. Free fluoride ions in the mineralizing milieu markedly affect the driving force for precipitation and, as a result, the nature of precipitating crystals. The fluoride incorporated into the crystalline lattice increases the stability of the formed mineral. Improvement in the understanding of the mechanism of dental fluorosis requires more comprehensive information about the effects of fluoride on the ionic composition of the fluid phase, the nature of the initially precipitating mineral(s), the interactions between crystals and matrix proteins, and the enzymatic degradation of the proteins. Recent observations relevant to the role of fluoride in enamel formation include: (1) that there are threshold concentrations of fluoride below which the precipitation and hydrolysis of thin-platy octacalcium phosphate is facilitated but beyond which de novo apatite precipitation prevails; (2) that the presence of fluoride in the mineralizing milieu most likely affects the steady-state concentrations of mineral lattice ions; (3) that incorporation of fluoride into the stable pool is retarded by the presence of matrix proteins, particularly amelogenins, which inhibit the growth of apatite crystals; (4) that increasing the degree of fluoridation of apatite crystals enhances the adsorption of amelogenins onto the crystal surface, and (5) that amelogenins pre-adsorbed onto apatite crystals are more resistant to enzymatic cleavages by trypsin (used as a prototype of amelogeninases).

Ameloblasts↗
PubMed1993

Characterization of the mineralization process in cultures of rabbit growth plate chondrocytes.

The present study was undertaken to investigate the mineralization process in chondrocyte cultures. Chondrocytes were isolated from the growth plate of ribs of 4-week-old rabbits. The nature and properties of mineral crystals precipitated in chondrocyte cultures were compared with those of crystals formed in the hypertrophic zone and bone of rabbit rib growth plates in vivo. The chondrocytes were maintained at high density on type II collagen-coated dishes in Eagle's medium, alpha-modification, with 10% fetal bovine serum and 50 micrograms/ml of ascorbic acid. These cells differentiated into hypertrophic cells 10 days after seeding and produced alkaline phosphatase and 1,25-dihydroxyvitamin D3 receptors on Days 30-70 at levels as high as those in the lower hypertrophic zone in vivo. Mineralization was initiated between Days 20 and 30 and advanced progressively throughout the culture period. However, mineralization was suppressed by the addition of parathyroid hormone (2 x 10(-8) M) or by the presence of fibroblasts. Examination by electron microscopy and Fourier transform infrared (FTIR) spectroscopy verified that mineralized nodules formed in vitro were composed of small apatite crystals. Importantly, FTIR spectral features of the apatite crystals (e.g., the prominent PO4 bands at 1125 and 1032 cm-1) were similar to those of cartilage apatites formed in vivo and differed markedly from those of carbonated bone apatites. These results suggest that growth plate chondrocytes cultured on collagen-coated dishes are an appropriate model for studies on cartilage mineralization.

Alkaline Phosphatase↗