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Column-like structures following the course of dentinal tubules in bovine, red deer, and rat teeth.

A material of bovine, red deer, and rat incisors was investigated, with emphasis on the bovine teeth. Both ground sections and paraffin wax sections of demineralized material were used. The sections were investigated by ordinary and polarized light microscopy and contact microradiography. In the bulk of the dentin irregular column-like light structures, alternating with largely similar, but dark, columns, followed the course of the dentinal tubules from the pulpal border to just below the mantle dentin. Each column, 10-100 micron in diameter, contained a highly varying number of tubules. The difference between light and dark columns was mainly related to the intertubular dentin of the globules of which they were composed. Light columns were slightly more mineralized than the dark ones and seemed to contain fewer GAGs, but more glycoproteins. A difference in fiber direction between light and dark columns was also evident. To the authors' knowledge these structures have not attracted the attention of previous workers.

Animals↗

Chemical and structural challenges in remineralization of dental enamel lesions.

The aim of the present paper was to identify some chemical and structural factors which may prevent a full remineralization of caries lesions and to study whether it is possible to overcome such obstacles. Samples of powdered enamel apatite were equilibrated with solutions metastably supersaturated with respect to enamel hydroxyapatite and fluorapatite. After 10 min and 60 min of equilibration at 20 degrees C the suspensions were centrifuged and the calcium and phosphate concentrations and the pH were determined in the supernatant. In parallel studies, 50 75-microns-thick sections of 27 fluorotic teeth of a severity of 5-7 according to Thylstrup and Fejerskov's classification were examined by microradiography and in polarized light using distilled water, and Thoulett's media or seen dry in air. Five obstacles inhibiting remineralization were identified: 1) Although remineralizing solutions or saliva are supersaturated with respect to enamel apatite the total amount of calcium and phosphate dissolved in it is small, so that after precipitation of the dissolved mineral only 1/20,000-1/30,000 of the volume of the mineralizing solution is occupied by mineral. 2) The concentration gradients from the mineralizing solution into the enamel is small, which indicates a slow diffusion into and out of the lesion. 3) The uptake of calcium and phosphate by the enamel apatite crystals is so rapid that the aqueous phase within the pores can be presumed to be only marginally supersaturated in the deeper parts of the lesion. 4) The surface layer of the enamel lesions was found to be a serious obstacle to remineralization so that a subsurface area remains hypomineralized after exposure to salivary remineralization even for a lifetime. 5) Nucleation of new apatite crystals to substitute lost crystals is an unsolved problem.

Apatites↗

A fiber-optic setup for quantification of root surface demineralization.

A fiber-optic fluorescence observation (FOFO) technique has been developed for the quantification of demineralized root dentin. The method was tested on 40 specimens of in vitro demineralized parts of human root dentin. Fluorescein sodium salt was used as a penetrating dye. The fluorescein sodium salt was excited using light around 465 nm. The fluorescence signal around 527 nm measured for demineralized dentin was corrected for lamp output and then divided by the corrected fluorescence signal for the sound control dentin on the same root, resulting in a FOFO-value. FOFO-values correlate linearly (r = 0.91) with mineral loss measured by transverse microradiography. Therefore, it was concluded that the FOFO-technique provides a good, non-destructive measure of the severeness of in vitro demineralized human root dentin, and is applicable on bulk dentin. Future work on natural lesions in vivo will be needed to investigate the applicability of the technique in a clinical situation.

Adolescent↗

Morphological appearance and chemical composition of enamel in primary teeth from patients with 22q11 deletion syndrome.

Patients with 22q11 deletion syndrome have many and complex medical problems, including hypocalcemia and/or hypoparathyroidism. Odontological findings include enamel aberrations in both dentitions. In order to describe enamel morphology, chemical composition in primary teeth, and to investigate the relationship between medical history and morphological appearance, dental enamel was investigated in 38 exfoliated primary teeth from 15 children and adolescents. Morphology was studied by the use of a polarized light microscope, microradiography, scanning electron microscopy, X-ray microanalysis, and secondary ion mass spectrometry. The morphological findings were compared with medical history. The teeth showed, in principle, a normal morphological appearance with regard to prism structure. A high frequency of aberrations, such as hypomineralization, hypoplasia and extra incremental lines, were found. The majority of the aberrations were found around the neonatal line. There was a relationship between high numbers of medical problems in the patients and enamel deviations. The result supports the hypothesis of under-reporting of both hypocalcemia and hypoparathyroidism in patients with 22q11 deletion syndrome.

Adolescent↗

Cataract in retinitis pigmentosa. An analysis of cataract surgery results and pathological lens changes.

The result of cataract surgery and lens pathoanatomical examination were presented in patients with retinitis pigmentosa (RP). A retrospective study of 24 cataract extractions in 15 RP patients was carried out, with special regard to visual outcome and surgical complications. The results were compared to an age matched randomly selected control group. In the retinitis pigmentosa group the post-operative visual acuity was generally good, and the surgical complications except for one case of central venous thrombosis were not more frequent than in the control group. Seven lenses from 5 patients with RP were analysed using transmission electron microscopy and quantitative microradiography. In the lenses with a progressive subcapsular edema, extensive extracellular vacuolization was found in the lens epithelium. Focal degeneration of epithelial cells was seen both in the more stationary posterior subcapsular cataracts and the progressive subcapsular cataracts. Different degrees of mitochondrial swelling was found in the epithelial cells of the stationary cataracts. The subcapsular lens fiber changes consisted of an extensive swelling of hydrated cells together with a significant reduction in their dry mass content. Migrating elongated nuclei-containing cells were found on the posterior capsule in both stationary and progressive cataracts.

Adult↗

Corneal alkali wound healing in the monkey.

Healing of a corneal alkali wound was followed in the monkey eye (Macaca fascicularis) for 4 months. A corneal alkali wound was inflicted in one eye in each of 30 monkeys under general anesthesia. A round filter paper, 5.5 mm in diameter, soaked in 1 N NaOH was put centrally on the cornea for 60 seconds. The alkali wounded all layers in the cornea underlying the filter paper, including the endothelium. Epithelial and endothelial healing was assessed morphometrically from photographs and micrographs, respectively. Stromal healing was documented using quantitative microradiography. During the observation period the wounds regained complete transparency. The epithelium showed few complications after primary healing during the first three days. The stroma regained normal thickness and dry mass content within one week. The endothelium resurfaced within one week and then remained intact. The role of the endothelial healing behaviour for the wound healing process is emphasized.

Alkalies↗

Exercise-induced pulmonary haemorrhage in the horse: results of a detailed clinical, post mortem and imaging study. VIII. Conclusions and implications.

This paper reviews a series of clinical, post mortem and imaging studies on exercise-induced pulmonary haemorrhage (EIPH) performed on 26 Thoroughbred racehorses. Post mortem techniques included routine gross, subgross and histological examination; coloured latex perfusions of pulmonary and bronchial circulations; and microradiography and computerised tomography scans of lungs with contrast injected vasculature. The major lesions were multiple, separate and coalescing foci of moderately proliferative small airway disease accompanied by intense neovascularisation of the bronchial circulation. As a result of bronchial artery angiogenesis, the systemic circulation dominated the vascular supply of the air exchange structures in affected areas, producing an apparent left to right shunt. Extensive areas of sequestered haemosiderophages indicated previous haemorrhage from vessels apparently supplied by the bronchial arteries. Diffuse and focal parenchymal destruction and connective tissue reactions in affected areas were considered to be secondary to localised haemorrhage and macrophage-induced damage. The aetiology of EIPH was not determined, but the multifocal, small airway-centred lesions indicated that low grade bronchiolitis, possibly of viral origin, was a factor. Gravitational effects also appear to contribute to dorsal distribution of the lesions. The mild focal and subclinical lesions confined to secondary lobules are thought to evolve into the serious lung pathology observed in EIPH cases through the effects of localised hypoxia induced by maximal exercise and partial airway obstruction. Once initiated, a vicious cycle of increasing inflammatory damage and further local bleeding is set in motion.

Animals↗

Remineralization of in vitro dental caries assessed with polarization-sensitive optical coherence tomography.

Polarization-sensitive optical coherence tomography (PS-OCT) is potentially useful for imaging the nonsurgical remineralization of dental enamel. This study uses an all-fiber-based PS-OCT system operating at 1310 nm to image demineralized and fluoride-enhanced remineralized artificial lesions. PS-OCT images of lesions before and after remineralization are compared with the relative mineral loss DeltaZ (%vol x microm), obtained from high resolution digital microradiography (DM), and chemical composition changes by infrared spectroscopy. Severe early artificial caries show a significant increase in perpendicular-axis integrated reflectivity after remineralization. After sectioning the samples, DM demonstrates that the lesions remineralized with new mineral and the lesion surface zone show significant restoration of mineral volume. PS-OCT and DM both do not show a major change in lesion depth. For less severe artificial caries, the perpendicular-axis image resolves the scattering and depolarization of an outer growth layer after remineralization. This outer layer has a mineral volume close to that of sound enamel, and spectroscopic analysis indicates that the layer is a highly crystalline phase of apatite, without carbonate substitutions that increase the solubility of sound enamel. This study determines that PS-OCT can image the effects of fluoride-enhanced remineralization of mild and severe early artificial in vitro caries.

Bone Density↗

Light scattering properties of natural and artificially demineralized dental enamel at 1310 nm.

A fundamental understanding of how near-IR light propagates through sound and carious dental hard tissues is essential for the development of clinically useful optical diagnostic systems, since image contrast is based on changes in the optical properties of these tissues on demineralization. During the caries (decay) process, micropores are formed in the lesion due to partial dissolution of the individual mineral crystals. Such small pores behave as scattering centers, strongly scattering visible and near-IR light. The optical properties of enamel can be quantitatively described by the absorption and scattering coefficients, and the scattering phase function. Our aim is to measure the optical scattering behavior of natural and artificial enamel caries. Near-IR attenuation measurements and angular-resolved goniometer measurements coupled with Monte Carlo simulations are used to determine changes in the scattering coefficient and the scattering anisotropy on demineralization at 1310 nm. An ultra-high resolution digital microradiography system is used to quantify the lesion severity by measurement of the relative mineral loss for comparison with optical scattering measurements. The scattering coefficient increases exponentially with increasing mineral loss. Natural and artificial demineralization increases the scattering coefficient more than two orders of magnitude at 1310 nm, and the scattering is highly forward directed.

Dental Caries↗

Predominant cultivable microflora of human dental fissure plaque.

Plaque developed in 10 occlusal fissures from unerupted third molars during implantation for 200 to 270 days in lower molars of dental students was studied. To characterize the predominant cultivable flora, 592 isolates (51 to 67 from each fissure) were subcultured from anaerobic roll tubes. Twenty-eight of the isolates were lost. Streptococci constituted 8 to 86% (median, 45%) of the isolates, Streptococcus mutans constituted 0 to 86% (median, 25%) and S. sanguis constituted 0 to 15% (median, 1%). A few isolates of "S. mitior" and "S. milleri" were found, but no S. salivarius. Staphylococci made up 0 to 23% (median, 9%). Gram-positive rods constituted 6 to 59% (median, 35%). Of these, 0 to 46% (median, 18%) were Actinomyces naeslundii and A. viscosus, but no anaerobic actinomyces were isolated. Arachnia and propionibacteria made up small proportions, lactobacilli were isolated from two fissures, constituting 10 and 29%, and eubacteria were isolated from one fissure (27%). Gram-negative cocci made up 0 to 46% (media, 4%). Only two isolates of gram-negative rods were found, both facultative anaerobes. Although 8 of the 10 fissures had large proportions of S. mutans, lactobacilli, or both, no caries was found even with microradiography. The large individual variation probably reflects differences in initial colonization from saliva and in growth conditions in each fissure.

Bacteria↗

Microflora associated with experimental root surface caries in humans.

This study describes the microflora from actively progressing root surface caries lesions, in which mineral loss had been determined by quantitative microradiography. The caries lesions were produced experimentally in root surface specimens from human molars inserted in lower partial dentures carried for 3 months by six elderly individuals. A total of 780 bacterial isolates were identified from 13 plaque samples, collected with a punch technique, and six dentin samples. The composition of the microflora showed distinct individual differences. The microflora from plaque samples associated with the highest mineral loss was dominated by either Actinomyces viscosus or a combination of mutans streptococci (serotypes c, d, and f) and Lactobacillus species (L. casei and L. brevis). Plaque from root surfaces with less pronounced mineral loss harbored a more complex microflora comprising gram-positive rods, mutans streptococci, Streptococcus mitis biovar 1, Veillonella spp., gram-negative rods, and low numbers of lactobacilli. In the latter samples, individual variations in the proportions of mutans streptococci (serotypes c, d, and g), Actinomyces species (A. viscosus and A. naeslundii), and Veillonella parvula biotypes were observed. These findings suggest that certain species or combinations of species are more cariogenic than others and that dominance of single acidogenic species in particular is conducive to high caries activity.

Aged↗

Osteoid osteoma: operative confirmation of complete removal by bone scintigraphy.

The high sensitivity of bone scintigraphy in detecting osteoid osteoma throughout the skeleton has been documented. Recently, radioactive surgical specimens of osteoid osteomas were studied with microradiography, autoradiography, and well-counter scintimetry and were shown to concentrate most of the radionuclide within the nidus of the lesion. These techniques also proved helpful to surgeons in achieving cure with conservative bone excision. We present two cases of osteoid osteoma, one of them recurrent, in which the patients were injected with bone-seeking agent just prior to surgery. The excised specimens were immediately imaged and the entire nidus was seen, confirming the complete removal of the lesion. This simple and rapid procedure is suggested whenever difficulty is anticipated in removal of an osteoid osteoma.

Adolescent↗

In vitro fluoride dose-response study of sterilized enamel lesions.

Currently our intra-oral model uses enamel specimens that have been disinfected by soaking in buffered formalin (pH 6.8). However, because of increasing emphasis on infection control, it is important to identify a way to sterilize these specimens. The aim of this study was to determine if autoclaved, or gas sterilized, lesioned enamel responds to fluoride (F) in the same way alcohol-disinfected enamel lesions do. Seventy-two formalin-disinfected, human enamel specimens (3 mm) were lesioned in demineralizing solution for 96 h and were then divided into three groups. One group was autoclaved; one group was gas sterilized (ethylene oxide), and the remaining 24 specimens were further disinfected in 70% ethanol for 10 min. Specimens in each group were then treated 4 times/day for 4 weeks with 0, 250 or 1,100 ppm F dentifrice slurries in an in vitro cycling, remin/demin model. Following treatment, fluoride uptake was analyzed by microdrill biopsy, and lesion depth and mineral content changes (DeltaM) were determined by transverse microradiography. Data were analyzed by one-way ANOVA analysis. In all three groups of specimens there were significant (p<0.05) differences in fluoride uptake in response to different fluoride treatments. Autoclaved lesions failed to provide dose- response data with regard to changes in lesion mineral content. Because formalin and 70% alcohol are only disinfectants, and autoclaving altered the responsiveness of enamel lesions, results from this study suggest that, of the methods tested, gas sterilization is the preferred method for sterilizing enamel specimens that will be used in intra-oral studies.

Analysis of Variance↗

Migration, matrix production and lamellar bone formation of human osteoblast-like cells in porous titanium implants.

The goal of this study was to characterize growth, mineralization and bone formation of osteoblast-like cells in titanium pore channels of defined diameter. Titanium implants with continuous drill channels of diameters of 300, 400, 500, 600 and 1,000 microm were inserted into human osteoblast-like cell cultures. The ingrowth of the cells into the drill channels was investigated by transmitted-light microscopy and scanning electron microscopy. Immunofluorescence and histological analysis of 15-channel sections of each diameter were used to investigate the growth behavior and the matrix protein patterns. Mineralization was evidenced by Alizarin red staining and high-resolution microradiography. The ingrowth of human osteoblast-like cells in the drill channels occurred in a sequence of four characteristic stages. In stage 1, osteoblast precursor cells adhered to the wall of the channel and migrated three-dimensionally into the channel by forming foot-like protoplasmic processes. For all 15 sample drill channels that were investigated, the cell ingrowth over 20 days amounted on average to 793 microm (+/- 179) into 600-microm-diameter channels, where they migrated significantly faster than in all the other channels. In stage 2, approximately on day 5-7, the osteoblast-like cells began to anchor on the substrate wall by matrix proteins and to build up a dense network of matrix proteins in the drill channel. The mineralization of the extracellular matrix, while depending on cell stimulation, was initiated in stage 3, on average after 4 weeks. In drill channels of a diameter of 1,000 microm the cell growth was incomplete and no mineralization was found by radiological assessment. Starting in week 6, in the drill channels of diameters ranging from 300 to 600 microm, the network of extracellular matrix proteins and osteoblast-like cells began to form an osteon-like structure. Neither the highly developed migration behavior of osteoblastic cells nor the reorganization from a fiber-like matrix to a lamellar structure have so far been described for cell cultures.

Bone Matrix↗

Acid resistance of enamel subsurface lesions remineralized by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate.

The aim of this clinical study was to investigate the acid resistance of enamel lesions remineralized in situ by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP: Recaldent). The study utilized a double-blind, randomized, crossover design with two treatments: (i) sugar-free gum containing 18.8 mg of CPP-ACP, and (ii) sugar-free gum not containing CPP-ACP as control. Subjects wore removable palatal appliances with insets of human enamel containing demineralized subsurface lesions and chewed the gum for 20 min 4 times per day for 14 days. After each treatment the enamel slabs were removed and half of each lesion challenged with acid in vitro for 8 or 16 h. The level of remineralization was determined using microradiography. The gum containing CPP-ACP produced approximately twice the level of remineralization as the control sugar-free gum. The 8- and 16-hour acid challenge of the lesions remineralized with the control gum resulted in 65.4 and 88.0% reductions, respectively, of deposited mineral, while for the CPP-ACP-remineralized lesions the corresponding reductions were 30.5 and 41.8%. The acid challenge after in situ remineralization for both control and CPP-ACP-treated lesions resulted in demineralization underneath the remineralized zone, indicating that the remineralized mineral was more resistant to subsequent acid challenge. The results show that sugar-free gum containing CPP-ACP is superior to an equivalent gum not containing CPP-ACP in remineralization of enamel subsurface lesions in situ with mineral that is more resistant to subsequent acid challenge.

Acetic Acid↗

Effect of saliva composition on experimental root caries.

The aim of this study was to determine the effect of saliva composition on caries lesion development independently of the flow rate of unstimulated whole saliva (UWS) and other caries-related variables such as lesion progression time, oral hygiene level, and fluoride exposure. We hypothesized that this could be done by developing experimental root caries under carefully controlled conditions in situ in test subjects with UWS flow rates within a narrow window of normalcy. Fifteen female and 5 male subjects (66 +/- 6 years) were selected for the study according to their UWS flow rates between 0.2 and 0.4 ml/min. All subjects developed experimental root caries lesions during a 62-day period in which UWS as well as stimulated whole saliva (SWS) were repeatedly collected and analysed for flow rate, pH, buffer capacity, inorganic, and organic composition. Caries lesion development was determined by quantitative microradiography. The mean UWS flow rate was 0.30 +/- 0.05 ml/min. Significant negative correlations were obtained between UWS total phosphate concentration and mineral loss (DeltaZ; r(s) = -0.72, p < 0.001) and UWS total protein concentration and DeltaZ (r(s) = -0.70, p < 0.01). SWS and its constituents had only limited or no effect on DeltaZ. Qualitative UWS protein analysis (SDS-PAGE) revealed that subjects with low DeltaZ values had broader and more stained amylase bands than subjects with high DeltaZ values. These findings were confirmed quantitatively by HPLC. We conclude that, within a group of subjects with normal UWS flow rates, the UWS composition was more important for caries lesion development than the SWS composition. Furthermore, high UWS concentrations of phosphate, protein, and amylase were caries-protective.

Aged↗

Caries-preventive agents induce remineralization of dentin in a biofilm model.

The aim was to study remineralization in dentin underneath a biofilm. This was done in a constant depth film fermentor (CDFF) which was modified so that two treatments can be applied simultaneously in one experiment. Forty-five Streptococcus mutans biofilms were grown in grooves in dentin. Growth medium (3.7 g/l BHI, 1.5 mM calcium and 25 mM PIPES) was administered alternately with 2% sucrose pulsing 4 x 30 min/day. Fluoride at 135 ppm as NaF only or in a mixture with 0.2% chlorhexidine was applied for 2 x 5 min/day. The treatments started 5 days after inoculation and lasted 15 days. Five specimens per group were removed at various time points. The biofilms were checked for viability (by plating) and acid content (by capillary electrophoresis). The dentin specimens were analysed for mineral loss and lesion depth (by transversal microradiography). Fluoride treatment had no effect on the viability but reduced lactic acid production by 75%. The mixture treatment reduced the viability by 80% and the lactic acid content by 93% on the first day and later reduced the two parameters to below the detection limits. Significant differences in changes in mineral loss and lesion depth were observed between the treatment groups. Partial remineralization but deeper lesions were observed in the fluoride group, while nearly complete remineralization was seen in the mixture group. In conclusion, the CDFF S. mutans biofilm model can be used as a de- and remineralization biofilm model, and the split mode is particularly suitable for testing caries-preventive agents.

Animals↗

Profilometric and microradiographic studies on the effects of toothpaste and acidic gel abrasivity on sound and demineralized bovine dental enamel.

The objective of this study was to assess the abrasive effects of toothpastes and acidic F gels on sound and demineralized enamel. Pairs of enamel specimens were cut from bovine incisors, embedded in epoxy resin and polished. An artificial subsurface lesion of 80-90 microm depth was created in one specimen from each pair. The samples were covered with adhesive tape, thereby exposing the enamel for abrasivity testing. All samples were divided into six groups of 15 and brushed with a slurry (1:3) of F gel or toothpaste and human saliva. Brushing with water (control) or with slurry was carried out (16,000 strokes) using a medium toothbrush (load 275 g) mounted in a brushing machine. Abrasion was evaluated using laser profilometry, and was about 50% less on sound than on demineralized enamel (p < 0.001). In the latter, brushing with water (0.09 +/- 0.03 microm) or with fluoride-free gel (0.08 +/- 0.03 microm) resulted in negligible wear. With a medium-abrasive paste (1.76 +/- 0.85 microm) and an acidic F gel (2.48 +/- 0.72 microm), brushing abrasion was significantly greater (p < 0.001) than with a low-abrasive paste (0.84 +/- 0.38 microm). The greatest wear (16.6 +/- 10.8 microm) was observed with high-abrasive paste (p < 0.001), and here transversal microradiography revealed a complete loss of the pseudointact surface after brushing. In vitro formed caries-like lesions can be abraded (by toothbrushing) more easily than sound enamel; hence, initial white spot lesions should preferably be brushed with oral hygiene products of low abrasivity.

Amines↗