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Effects of dentin proteins, transforming growth factor beta 1 (TGF beta 1) and bone morphogenetic protein 2 (BMP2) on the differentiation of odontoblast in vitro.

We have studied the effects of dentin proteins, of Transforming Growth Factor beta 1 (TGF beta 1) and Bone Morphogenetic Protein (BMP2) on the differentiation of odontoblasts in vitro. The total EDTA-soluble fraction of dentin proteins, prepared from rabbit incisors was further separated by chromatography on DEAE-Cellulose and heparin-agarose columns. While the total EDTA-soluble fraction of dentin had no effect on cultured dental papillae, fractions retained on both columns were able to initiate functional differentiation of preodontoblasts of isolated day-17 first lower mouse molar dental papillae cultured in vitro. TGF beta 1 and BMP2, both stimulated the matrix secretion by dental papillae cells. TGF beta 1 and BMP2, combined with the inactive total EDTA-soluble fraction, stimulated odontoblast differentiation. An active fraction retained on DEAE-Cellulose completely lost the inductive activity after incubation with a neutralizing anti-TGF beta antibody. These results demonstrate that a TGF beta-like molecule present in dentin could interact with some component which acts as a modulator of its activity on the initiation of the cytological and functional differentiation of odontoblasts.

Animals↗

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↗

Fluoride retention in dentin after topical application of aminefluoride.

The retention of KOH-soluble and structurally bound fluoride in dentin after topical application of aminefluoride was investigated. One cylindrical dentin sample was prepared from each of 40 extracted third molars. All samples received one topical application of 1 microL of aminefluoride solution (1.23% F) for three min. Ten of them were analyzed immediately after fluoridation. The remaining 30 samples were mounted in the buccal aspects of a special mouth application which was carried by a person for five days. After one, three, and five days, 10 of the 30 samples were removed from the appliance. In all samples, the amount of structurally bound fluoride was determined in three layers (20 microns, 20 microns, 20 microns) by use of a special grinding technique. Before each grinding step, the dentin specimens were analyzed for KOH-soluble fluoride. The total amount of KOH-soluble fluoride in dentin was 48.7 +/- 14 micrograms/cm2. It decreased drastically during the experimental period. Structurally-bound fluoride increased one day after fluoridation in the first and second layers. Thus, topical application of aminefluoride resulted in a significant accumulation of KOH-soluble fluoride in dentin. This precipitate was unstable, but fluoride ions released from it could perhaps increase the amount of structurally bound fluoride.

Amines↗

[Insolubilization of deciduous tooth enamel by the two-step method with fluoride and lanthanum treatment].

The acid-resistance of the tooth surface is strengthened to form noncrystal lanthanum fluoride on its surface by the two-step treatment (F-La treatment) with acidulated-phosphate fluoride and lanthanum chloride solutions as previously reported by Fujiwara, Negishi, Miyagi et al. This study has been made to examine some clinical applications of the treatment. The acid-resistance effect and the actual condition of the F-La treatment were investigated on the extracted human deciduous dentin. Analysis was made using the electron probe X-ray microanalyzer before and after the acid solubility test. The results were as follows: 1) With F-La treatment, the acid-solubility of deciduous tooth dentin was reduced to about one-third of that without treatment, and to about two-third compared both with F-treatment and with diamine silver fluoride application. 2) Applications of 3 to 5 minutes with F and 3 minutes with 0.5 to 1.0% La sufficed for this F-La treatment. 3) An etching procedure was not necessary previous to the treatment in contrast with the application to the enamel. 4) Fluorine and lanthanum penetrated up to approximately 10 microns from the dentin surface with this treatment. Even after the acid-solubility test, a sufficient quantity of the stated chemicals remained, thus protecting the tooth substance. 5) The acid-solubility of the deciduous dentin was higher than that of the deciduous enamel. After the F-La treatment, however, the former diminished more markedly than the latter. Based on these experimental results, one can confirm the fact the chemical reactivity of the deciduous dentin is higher than that of the deciduous enamel.

Acidulated Phosphate Fluoride↗

[On the acid solubility of the dental enamel of rabbits after internal administration of various antibiotic substances (author's transl)].

Sections of etched enamel and dentinal surfaces were examined to compare the acid solubility of normal hard tissue and determine the effects of various antibiotic substances. Considerable differences were observed, of which the correlation with cariogenesis and with the progression of caries are being discussed in detail. The results of this study show that treatment with antibiotics may lead to reductions in quality in infants, young children, and nursing mothers.

Acid Etching, Dental↗

[Effect of sodium chloride treatment on remineralization of human root dentine caries in vitro].

OBJECTIVE: To investigate the influence of soluble phosphoprotein removal from artificial dentin caries lesions by pretreatment of NaCl on subsequent remineralization. METHODS: Human root dentin samples were demineralized in an acidic gel (pH = 4.4) at 37 degrees C for 48 hours. Samples were pretreated with 0.5 mol/L NaCl solution and 0.5 mol/L disodium EDTA solution respectively. The histomorphologic changes of root caries surface were examined by scannings electronic microscope. Mineral profiles were assessed by means of microradiography. RESULTS: No histomorphologic changes were present on root caries surface after treated with 0.5 mol/L NaCl solution, while many opened dentin tubules were seen after treated with 0.5 mol/L disodium EDTA solution. The remineralization results showed that pretreatment with 0.5 mol/L NaCl solution increased more remineralization than pretreated with 0.5 mol/L disodium EDTA solution. CONCLUSION: 0.5 mol/L NaCl solution has no damage on the root caries surface. The in vitro results suggested that removal of soluble phosphoprotein from dentin lesions is an interesting approach to enhance remineralization.

Dental Caries↗

Age estimation from racemization rate using heated teeth.

The racemization ratio (D/L ratio) of aspartic acid was determined in soluble peptide and total amino acid extracted from the dentine of experimentally-heated and unheated teeth. The ratio was higher in heated than in unheated teeth for total amino acid, but approximately the same for soluble peptide. Although the estimated age for total amino acid was much greater than the true age for heated teeth, it was approximately the same for total amino acid in unheated teeth and for soluble peptide in heated teeth and unheated teeth. These results suggest that age calculated from the D/L ratio of dentinal soluble peptide from burned bodies is sufficiently accurate for this parameter to be used to estimate age in teeth from burned bodies.

Adult↗

Molecular biological studies on teeth, and inquests.

Reviewed here are three dental approaches to identification work. One is that using racemization of amino acids. Age estimation of a high correlation of 0.99 was achieved from using the whole dentine from the central vertical section. The highest correlation was also seen when soluble peptide from dentine was used, suggesting its usefulness for age estimation. The racemization rate was highest for Asp, followed by Glu and Ala in that order. The other approach reviewed in DNA typing. DNA is extracted from dental pulp for DNA fingerprinting with Y-specific probe. DNA typing is especially useful for sex determination, allowing valid determination from teeth extracted up to 21 months before the examination. A 3-cycle repetition of PCR provided an accurate sex determination from a considerably degraded DNA specimen, comparable to one freshly sampled. The other approach discussed is the content and scope of the inquests performed by dentists who participate in the identification work in mass disasters etc. In this context, the antemortem dental information including dental records and radiological records provided by families of the victims, play a major role in speedy identification.

Age Determination by Teeth↗

Efficacy of a dentifrice containing potassium nitrate, soluble pyrophosphate, PVM/MA copolymer, and sodium fluoride on dentinal hypersensitivity: a twelve-week clinical study.

The effect on dentinal hypersensitivity from the use of a dentifrice containing 5.0% potassium nitrate, 1.3% soluble pyrophosphate, 1.5% PVM/MA copolymer, and 0.243% sodium fluoride in a silica base over a twelve-week period was compared to a matching placebo dentifrice without potassium nitrate. A total of sixty-seven subjects were entered into the study, and stratified into two balanced groups according to their baseline mean thermal (threshold) temperature (Thermodontic Stimulator) and baseline mean tactile (Yeaple Probe) sensitivity scores. The two groups were randomly assigned to use either the potassium nitrate/copolymer/pyrophosphate dentifrice or the placebo dentifrice. The two groups were well balanced with regard to their mean baseline thermal and tactile sensitivity scores, sex and age. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice and a commercially available soft-bristled toothbrush. Dentinal hypersensitivity examinations, which included tactile sensitivity, threshold thermal sensitivity, pain thermal sensitivity, air blast, and a visual analog scale were conducted at baseline, six weeks, and twelve weeks. All examinations were conducted by the same dental examiner. After six weeks' use of their assigned products, those subjects in the potassium nitrate/copolymer/pyrophosphate dentifrice group demonstrated statistically significant improvements (p < 0.01), as compared to the placebo dentifrice, in the following parameters: 1) tactile; 2) thermal (threshold and pain); and 3) air blast. After twelve weeks' use of their assigned products, those subjects in the potassium nitrate/copolymer/pyrophosphate dentifrice group again demonstrated statistically significant improvements (p < 0.01), in tactile, thermal (threshold and pain) and air blast sensitivity, as compared to the placebo dentifrice. It was concluded from this study that a dentifrice containing 5.0% potassium nitrate, 1.3% soluble pyrophosphate, 1.5% PVM/MA copolymer, and 0.243% sodium fluoride in a silica base is a clinically effective treatment for reducing dentinal hypersensitivity.

Adult↗

Protective effects of cavity liners on dentin.

Smear layers are very soluble in acidic solutions. Removal of smear layers increases dentin permeability and the potential for pulpal irritation. In this study, smear layers were treated with either saline, Barrier, Copalite, LC-Scotchbond, Hydroxyline, or DDS 1 & 2 to determine how well they protected the dentin from subsequent acid attack with 37% phosphoric acid for two minutes. Using both SEM and permeability measurements, the materials were ranked from least to most effective as: Saline less than Barrier = Copalite less than Hydroxyline = Scotchbond = DDS 1 & 2.

Acid Etching, Dental↗

Current concepts on the theories of the mechanism of action of fluoride.

The caries-preventive effect of fluoride is mainly attributed to the effects on demineralization/remineralization at the tooth oral fluids interface. Sub ppm levels of fluoride in saliva are effective in shifting the balance from demineralization, leading to caries, to remineralization. This is attributed to the fluoride-enhanced precipitation of calcium phosphates, and the formation of fluorhydroxyapatite in the dental tissues. Low fluoride levels are found in saliva after toothbrushing with fluoride containing dentifrices. Similar concentrations are ineffective in interfering with processes of growth and metabolism of bacteria, and also do not result in a significantly reduced dissolution of tooth mineral as a result of (firmly bound) fluoride incorporation. Comparative studies of fluoride efficacy have shown that higher concentrations in solution are needed in pH-cycling studies of dentine than in enamel to maintain the mineral balance or to induce remineralization. This is attributed to the greater solubility of the dentine and the smaller size of the dentine crystallites compared to enamel. Fluoride slow-release devices, in the form of fluoride-releasing restorative materials, may serve to increase the fluoride levels in saliva and plaque to levels at which caries can be prevented, also in high-risk patients. Research questions for the next millennium and future perspectives for fluoride applications should be found in the retention and slow release of fluoride after various combinations of fluoride treatment, the combination of fluoride and anti-microbial treatment, the individualization of caries prevention, and the combination of preventive schemes with new developments in caries diagnosis.

Cariostatic Agents↗

Reactionary dentinogenesis after applying restorative materials and bioactive dentin matrix molecules as liners in deep cavities prepared in nonhuman primate teeth.

OBJECTIVE: The aim of this in vivo study was to evaluate the response of the pulp-dentin complex following application of resin-modified glass-ionomer cement, calcium hydroxide hard-setting cement and EDTA-soluble preparation of dentine matrix proteins (ESDP) in deep cavities prepared in non-human primate teeth. METHODS: Eighteen deep Class V buccal cavities were prepared in premolars of four capuccin monkeys. In Groups 1 and 2, the cavity floor was lined with ESDP or a resin-modified glass-ionomer cement (Vitrebond - 3M ESPE), respectively. In Group 3 (control), the cavity was lined with a hard setting calcium hydroxide cement (Dycal - Dentsply). The cavities were subsequently filled with amalgam. After 6 months, the animals were sacrificed and the teeth were prepared for microscopic assessment. Six-micron thick serial sections were stained with H/E, Masson's trichrome and Brown & Brenn techniques. RESULTS: No inflammatory pulpal response was observed for all experimental and control Groups. However, the amount of reactionary dentin deposition differed between groups in the rank order ESDP (Group 1) > calcium hydroxide (Group 3) > resin-modified glass-ionomer (Group 2). These differences were statistically significant. CONCLUSIONS: All materials were biocompatible when applied in deep cavities. ESDP stimulated higher deposition of reactionary dentin matrix than Vitrebond and Dycal.

Animals↗

Study on acid resistance of human dental enamel and dentin irradiated by semiconductor laser with Ag(NH3)2F solution.

OBJECTIVE: The purpose of this study was to investigate the acquired acid resistance of human dental enamel and dentin treated by 38% diamine silver fluoride [Ag(NH3)2F] solution and semiconductor (diode) laser irradiation in vitro. BACKGROUND DATA: There have been no reports on the acid-resistant effect by combined use of fluoride and semiconductor laser. METHODS: Sixty crowns of extracted human molars were divided into two groups for enamel and dentin samples, and each group was subdivided into three subgroups of 10 each. Each subgroup of enamel and dentin samples served as a control; one was treated with Ag(NH3)2F and the other was treated with Ag(NH3)2F and semiconductor laser irradiation at 2 W for 30 sec. Then all samples were immersed in 5 ml of 0.1 M lactic acid (pH 4.8) at 37 degrees C for 24 h. The concentration of calcium ion (Ca2+) dissolved in the solution was measured by atomic absorption spectrophotometry, and the samples were observed by stereoscopy and scanning electron microscopy (SEM). RESULTS: In both enamel and dentin samples, dissolved Ca2+ concentration in Ag(NH3)2F- or Ag(NH3)2F- and laser-treated group was significantly decreased compared with that in the control (p < 0.01). SEM findings showed that numerous cubic particles ranging from 0.1 to 0.5 mcirom were observed only in the combined treated groups of both enamel and dentin samples. CONCLUSIONS: A significantly decreased solubility of human enamel and dentin was acquired after treatment by Ag(NH3)2F and semiconductor laser irradiation, which suggested that this combined use has the capability of a more efficient acid-resistant effect on human dental hard tissues.

Cariostatic Agents↗

An extracellular matrix protein of dentine, enamel, and bone shares common antigenic determinants with keratins.

EDTA-soluble proteins extracted from rabbit dentine were injected into mice and antibodies were prepared by the hybridoma technique. A monoclonal antibody, MC9B5, was found to react with a 68 kd protein, a major component of the EDTA-soluble proteins present in rabbit and rat dentine. Weston immunoblotting demonstrated that the antigen was also present in rat enamel and bone. By indirect immunofluorescence, MC9B5 was found to specifically stain epithelial filamentous cytoskeletal structures. Double staining experiments using MC9B5 and antikeratin antibodies showed extensive co-localization on the epithelial cell cytoskeleton. Furthermore, the 68 kd extracellular matrix protein was recognized in dentine, enamel, and bone extracts by antikeratin antibodies as shown by immunoblotting. These data support a structural relationship between a mineralized tissue extracellular protein and keratins.

Animals↗

Fluoride levels in dentin after iontophoresis of 2% NaF.

The fluoride in successive layers of dentin (from 3-6 micron were removed per etch) has been measured following topical or iontophoretic application of 2% aqueous sodium fluoride to coronal exposed dentin of freshly extracted human third molar teeth. The fluoride contents of the topical and iontophoresed first etches were, respectively, 12 and 22-31 times greater than that of control (untreated) teeth. Similarly, the fourth etches were 12 and 50-70 times greater, and the seventh (last sampled) etches were 5 and 28-41 times greater. There was no apparent change in acid solubility of the dentin as its fluoride content varied under the conditions of this study. This is the first report studying the fluoride content of extremely thin layers of dentin successively removed by acid etch and relating topical to iontophoretic uptake.

Acid Etching, Dental↗

In vivo morphogenetic activity of dentine matrix proteins.

Matrix proteins (EDTA-soluble and collagenase-released) from rabbit dentine have been demonstrated to show in vivo morphogenetic activity in exposed pulps of ferret canine teeth. In young adult ferrets, the reparative dentine resembled normal primary tubular dentine with odontoblast-like cells lining its formative surface. The reparative response in older animals was seen as osteo-dentine deposition. In both cases, the reparative response was characterized by its intensity within a comparatively short time period and also by the low grade or complete absence of any localized inflammatory response. Dentine matrix proteins implanted in exposed pulps of teeth provide a good experimental model for analysing the cell-matrix interactions and events taking place during odontoblast differentiation and dentinogenesis.

Aging↗