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Mineral element analysis of carious and sound rat dentin by electron probe microanalyzer combined with back-scattered electron image.

We recently demonstrated the advantages of back-scattered electron images (COMPO) in the visualization of dentinal caries, and the relationship of the change in the dentin fluorescence pattern in caries lesions. However, the exact nature of these changes is not known. In this paper, the nature of the changes in the areas with reduced mineral content in COMPO images was investigated. We examined the relation of changes in mineral elements and the appearance of soft carious and sound dentin in COMPO images using a scanning electron microscope (SEM) equipped with an electron probe microanalyzer (EPMA). Rat molars with small dentinal caries lesions just under the DEJ were chosen for the study. The Ca, P, Na, Mg, Zn, F, and total contents were determined by EPMA from five different dentin sites, and the Ca/P and Mg/Ca ratios were calculated. Generally, the lowest contents were found in caries lesions and highest in mantle dentin, with the exceptions of Mg and Zn. The Ca/P ratio was lowest in mantle dentin and highest in carious dentin. The results confirm that the change in fluorescence in the dentinal caries lesion is correlated with the very initial changes in mineral content, and that EPMA used in combination with COMPO images is a useful tool for determining small changes in mineral elements in the carious and adjacent areas of dentin.

Animals↗

Transmission electron microscopy of cementum crystals correlated with Ca and F distribution in normal and carious human root surfaces.

Root-surface caries, like enamel caries, develops as a subsurface type of mineral loss. Very little is known about the composition of the surface zone covering the body of the lesion, and the ultrastructure and composition of carious cementum are not known. The aim of this study was to correlate the ultrastructure and arrangement of the cementum crystals with the distribution of fluoride and calcium in root cementum from human teeth with sound, unexposed, or exposed root surfaces as well as in early stages of root-surface caries. Microradiographically, unexposed specimens showed a relatively homogeneous mineral distribution contrasting with the formation of an apparently highly mineralized surface layer in exposed and, in particular, in carious cementum. The electron-probe findings showed a substantial fluoride peak corresponding to the surface layers in carious tissues in particular, whereas the calcium profile in the surface did not reflect the apparent increase in mineralization. A substantial increase in size of the cementum crystals was found in specimens with formation of the fluoride-rich, well-mineralized surface zone. The crystal lattice intervals when observed along the (001) plane showed a hydroxyapatite spacing. The findings indicated that a significant crystal growth can be achieved in human cementum concomitant with fluoride accumulation.

Adult↗

Ultrastructural properties of laser-irradiated and heat-treated dentin.

Previous studies using scanning electron microscopy and infrared absorption spectroscopy reported that laser irradiation causes compositional changes in enamel. The purpose of this study was to evaluate the ultrastructural and compositional changes in dentin caused by irradiation with a short-pulse laser (Q-switched Nd:YAG). The irradiated and non-irradiated areas of the lased dentin samples were investigated by scanning (SEM) and transmission electron microscopy (TEM), micro-micro electron diffraction, and electron microprobe analysis of dispersive energy (EDX). Heat-treated dentin was similarly investigated. This study demonstrated that laser irradiation resulted in the recrystallization of dentin apatite and in the formation of additional calcium phosphate phases consisting of magnesium-substituted beta-tricalcium phosphate, beta-TCMP, beta-(Ca,Mg)3(PO4)2, and tetracalcium phosphate, TetCP, Ca4(PO4)O. TEM analyses of the modified and unmodified zones of the irradiated areas showed two types of crystal populations: much larger crystals from the modified zone and crystals with size and morphology similar to those of dentin apatite in the unmodified zone. The morphology of crystals in the modified zones in the irradiated dentin resembled those of dentin sintered at 800 or 950 degrees C. In the irradiated areas (modified and unmodified zones), the Ca/P ratio was lower compared with that in the non-irradiated dentin. The Mg/Ca ratio in the modified zones was higher than that in the unmodified zones and in the non-irradiated dentin. In sintered dentin, the Mg/Ca ratio increased as a function of sintering temperature. The ultrastructural and compositional changes observed in laser-irradiated dentin may be attributed to high temperature and high pressure induced by microplasma during laser irradiation. These changes may alter the solubility of the irradiated dentin, making it less susceptible to acid dissolution or to the caries process.

Calcium↗

Antibacterial activity of zinc modified titanium oxide surface.

Titanium-based implants are successfully used for various biomedical applications. However, in some cases, e.g. in dental implants, failures due to bacterial colonization are reported. Surface modification is a commonly proposed strategy to prevent infections. In this work, titanium oxide, naturally occurring on the surface of titanium, was modified by promoting the formation of a mixed titanium and zinc oxide, on the basis of the idea that zinc oxide on titanium surface may act as the zinc oxide used in pharmaceutical formulation for its lenitive and antibacterial effects. The present work shows that it is possible to form a mixed titanium and zinc oxide on titanium surfaces, as shown by Scanning Electron Microscopy and XPS analysis. To this end titanium was preactivated by UV on crystalline titanium oxide, both in the anatase form or in the co-presence of anatase and rutile. By performing antibacterial assays, we provide evidence of a significant reduction in the viability of five streptococcal oral strains on titanium oxide surfaces modified with zinc. In conclusion, this type of chemical modification of titanium oxide surfaces with zinc might be considered a new way to reduce the risk of bacterial colonization, increasing the lifetime of dental system applications.

Anti-Bacterial Agents↗

Mineralogical analysis of the respiratory tract in aluminium oxide-exposed workers.

A retrospective study was conducted in order to characterize the retention of fibrous and nonfibrous mineral particles in the respiratory tract in subjects with previous occupational exposure in the aluminium industry. Bronchoalveolar lavage (BAL) fluid (three samples) or lung parenchyma (two samples) were studied using analytical transmission electron microscopy in five patients. A high concentration of aluminium fibres (> 10(7) fibres.g-1 dry lung) was identified in two lung tissue samples, and aluminium fibres were also identified in BAL fluid in three patients. All fibres were short (mean length: 1-2 microns), with no fibre longer than 5 microns. Some biopersistence of these fibres in the respiratory tract is suggested from these observations, since fibres were identified in biological samples collected more than 4 yrs after cessation of exposure in four out of five patients. Occupational physicians should be aware of possible exposure to short, thin aluminium fibres during primary aluminium production. Further studies are needed to assess the potential health effects of these fibres. Industrial hygiene measurements should also be performed to document the potential sources of exposure to aluminium fibres in this industry.

Adult↗

Elemental modifications and polycyclic aromatic hydrocarbon metabolism in human fibroblasts.

Mineral fibers and particulates represent one of the best documented, economically important, and ubiquitously occurring categories of human carcinogens. Yet, while a wealth of information exists concerning the mechanism of action of physical, chemical, and viral carcinogens, virtually nothing is known relative to the mechanism of action of this economically important class of carcinogenic compounds known as mineral fibers and particulates. While the length and diameter of various forms of asbestos have been associated with both cellular toxicity in vitro and tumor occurrence in vivo, nothing is known about whether or not these same physical properties are responsible for the purported synergistic interaction between cigarette smoking and asbestos exposure relative to the induction of bronchogenic carcinoma. Thus, while the risk of bronchogenic carcinoma for nonsmokers exposed to asbestos appears to be only slightly greater than that for unexposed nonsmoking populations, the risk of occurrence of this same tumor in asbestos workers who also smoke is approximately 100-fold greater than in nonsmoking asbestos workers. The risk of bronchogenic carcinoma is increased approximately 8-fold in asbestos workers who smoke over non-exposed smokers. Since it is clear that cigarette smoke contains over 150 polycyclic aromatic hydrocarbons, some of which are known animal carcinogens, and since other laboratories have reported that metals associated with asbestos redirect the metabolism of these agents, it was of interest to us to investigate the effects of mineral fibers on the metabolism and biochemistry of polycyclic aromatic hydrocarbons.

9,10-Dimethyl-1,2-benzanthracene↗

Effect of zinc upon cell attachment and proliferation on periodontally-diseased root surfaces.

The purpose of this study was to determine the effect of zinc ions on fibroblast attachment to periodontally-diseased root surfaces in vitro. Extracted periodontally-diseased teeth were treated with 0.5% ZnCl2 by iontophoresis at 0.5 mA for 2 to 6 minutes. Control groups were untreated diseased and untreated healthy teeth. Sections of the root underlying the pocket were cut from the diseased teeth. The specimens were incubated for 18 hours with L929 mouse fibroblasts, then transferred and incubated in cell-free medium for 48 hours. Cell attachment was evaluated by cell counts and scanning electron microscopy (SEM). Root surfaces were sampled with an acid-etch technique and zinc was measured with an atomic absorption spectrophotometer. Two samples from each group were examined for cell attachment with SEM. Data were analyzed using the appropriate statistical methods. The results showed that diseased, untreated root surfaces had significantly fewer cells attached; however, zinc iontophoresis did not significantly improve cell attachment to the diseased root surfaces. Zinc analysis showed that diseased, untreated root surfaces had a higher zinc content than healthy ones. SEM examination showed striking differences in cell attachment to healthy versus diseased root surfaces. The data indicated that zinc iontophoresis did not significantly enhance cell attachment to root surfaces of diseased teeth.

Cell Adhesion↗

Root surface characteristics associated with subgingival placement of monolithic tetracycline-impregnated fibers.

The purpose of this investigation was to inspect and characterize the subgingival root surface after a 10-day exposure in vivo to 25% tetracycline hydrochloride by weight in an ethylene vinyl acetate copolymer fiber matrix with and without root planing therapy. The root surfaces were examined by fluorescent-light microscopy (FLM), scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS). Thirty-two teeth were selected for study, 4 from each of 8 patients. The teeth of each patient were randomly assigned to one of four treatment groups: non-treated control (C), scaling and root planing only (RP), tetracycline-impregnated fiber only (F), and scaling and root planing with tetracycline-impregnated fiber application (RP/F). SEM revealed a visible reduction in the subgingival microbial flora in both the F and RP/F treatment groups in comparison with the C group specimens. Many of the residual bacteria observed in F and RP/F specimens appeared non-viable, exhibiting obvious loss of membrane integrity. In contrast, the RP specimens exhibited randomly distributed areas of residual subgingival plaque and calculus with newly developing plaque fronts; the plaque fronts undoubtedly having formed during the 10 days post-therapy. All RP/F specimens exhibited an incomplete removal of adsorbed root surface pellicle and demineralization of the subsequently exposed root surface. EDS analysis of large crystals adhering to root surfaces of F and RP/F specimens revealed high chloride peaks, suggesting the presence of residual tetracycline. FLM examination of F and RP/F treated specimens showed a superficial penetration of tetracycline into the root surface of about 10 microns. Areas of demineralized root showed slight tetracycline penetration into exposed dentinal tubules.

Adsorption↗

Effect of a ceramic and a non-ceramic hydroxyapatite on cell growth and procollagen synthesis of cultured human gingival fibroblasts.

BACKGROUND: Ceramic hydroxyapatites and non-ceramic hydroxyapatites have been used extensively as alloplastic materials for bone reconstruction. However, different forms of hydroxyapatite induce different types of tissue response. METHODS: Human gingival fibroblasts (FMM1 cells) were used to analyze ceramic and non-ceramic hydroxyapatite biocompatibility. The cells were grown on surfaces covered either by collagen (control group), collagen plus ceramic hydroxyapatite, or collagen plus non-ceramic hydroxyapatite. Scanning electron microscopy, growth and cell viability curves, and procollagen immunoprecipitation were obtained. For the growth and viability curves, 10(4) cells were seeded on 60 mm dishes. Cells from each group were counted, in triplicate, at 1, 3, 4, 5, and 6 days after seeding using the Trypan blue dye exclusion assay. RESULTS: The cells grew in close contact with both types of hydroxyapatite particles. No differences were found in the amount of procollagen synthesis among any experimental group. The cultures treated with ceramic hydroxyapatite had a growth delay for the first 5 days. There was no difference in cell viability between the control group and the non-ceramic hydroxyapatite group. However, cultures treated with ceramic hydroxyapatite showed significantly lower viability percentages than the other groups. CONCLUSIONS: Hydroxyapatite supports cell growth and fibroblast metabolism including collagen production, and hence is biocompatible. Cell viability and structural studies showed that non-ceramic hydroxyapatite has relevant physical and biological properties as an implant material.

Algorithms↗

Internal contamination of a 2-component implant system after occlusal loading and provisionally luted reconstruction with or without a washer device.

BACKGROUND: Microbial internal contamination of oral implants has been suggested as being responsible for inflammatory infiltration of peri-implant tissues at the fixture-abutment interface of 2-stage and 2-component implant systems; it is also considered as a potential source of pathogens. The present in vivo study evaluated contamination of the inner abutment-implant retaining screw, after occlusal loading and provisionally-luted reconstruction with or without a washer device. METHODS: Eight 2-component oral implants sealed with an o-ring silicon washer device and 9 without a seal were placed in 7 patients with high oral hygiene standards. Two months after prosthetic reconstruction, crowns and internal screws were removed, and organic and inorganic screw contamination was examined by scanning electron microscopy and energy dispersive x-ray spectroscopy (EDS) analysis. RESULTS: An amorphous and crystalline contamination, suggestive of calcium and phosphate compounds, was seen on all screw surfaces. Microbial contamination was more frequently observed in the unsealed group. No differences in bacterial morphotypes were observed between the sealed and unsealed implants. Cocci were the most representative morphotypes, while rods were seldom seen. CONCLUSIONS: In clinical situations, leakage occurs at the implant-abutment interface, although bacterial contamination is limited in patients with high oral hygiene standards. Contamination may be reduced by using a washer device.

Analysis of Variance↗

The absence of correlations between a clinical classification and ultrastructural findings in amelogenesis imperfecta.

This study was performed to examine whether a clinical classification of different phenotypes of amelogenesis imperfecta could be discernible at the ultrastructural level. Seventeen primary teeth from 16 children with hypomineralization, hypomaturation, or hypoplastic variants of the disease were collected for histologic studies of the enamel by means of polarized light microscopy, scanning electron microscopy (SEM), and secondary ion mass spectrometry (SIMS). Polarization microscopy showed that the enamel was hypomineralized; in six teeth a wavy configuration of the enamel prisms also appeared. Three histomorphologic main types could be discerned. In 10 of the teeth extensive hypomineralization of the bulk of the enamel was found. One tooth had an unusually thick enamel with only a thin normally mineralized surface layer. SIMS images showed less pronounced signals from Ca2+ and Na+ but with stronger signals from Cl- and CN-, representing the organic component of enamel. The SEM images showed an irregular prism pattern with marked interprismatic areas. Irrespective of the clinical appearance or the hereditary pattern the main findings were hypomineralized enamel with or without wavy bands. Neither of the analytical methods used in this paper distinguishes between the clinical phenotypes of amelogenesis imperfecta.

Amelogenesis Imperfecta↗

Development of hereditary tapetal degeneration in the beagle dog.

Laboratory beagle dogs with an apparent absence of a tapetum lucidum were identified by ophthalmoscopic examination. Breeding experiments demonstrated a probable autosomal recessive mutation. Studies of the development of the tapetal abnormality showed that up to postnatal day 21 the tapetum was normal by light and ultrastructural morphology. Subsequent to that time the tapetal rodlets failed to accumulate electron-dense material, did not accumulate zinc, and degenerated primarily into spherical inclusion bodies of varying electron density. In the early phases of the degeneration the rough endoplasmic reticulum formed large whorls of membrane denuded of ribosomes. With time, the inclusions became electron lucent, and the entire tapetal cell degenerated, ending with almost total loss of the tapetum lucidum by approximately one to two years of age. The structure of the retina was normal. Retinal function measured by electroretinography was normal except for a slight elevation of dark adapted white light thresholds. It is speculated that the hereditary defect may be defective synthesis of the tapetal rodlet matrix or of the zinc-complexing substance of the tapetum.

Animals↗

In vivo wear pattern of experimental composite resins based on different resin monomers.

This study investigated the effects of various monomer systems on composite resin wear in vivo. Experimental light-cured composite resins were prepared employing four different monomer systems: (1) Bis-GMA type, (2) D-2. 6E type, (3) UDMA type, (4) UTMA type. The resin monomers consisted of 70wt% main monomer and 30wt% TEGDMA. These composites contained 80wt% fine quartz. The resins were placed in 2 mm diameter cylindrical cavities located in the occlusal contact area or the contact free area in cast crowns, temporarily set in a mouth. The crowns were removed at monthly intervals, for longitudinal SEM observation. Two months after setting, wear was analyzed, using an electron probe surface roughness analyzer. Microabrasion of the resin matrix and loss of filler particles were observed for all types of monomer systems. The effect of matrix resin systems on occlusal wear was smaller than that of filler systems.

Bisphenol A-Glycidyl Methacrylate↗