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Contact microradiography of dentine under wet conditions to prevent lesion shrinkage.

A considerable shrinkage of dentinal lesions is the result of desiccation of dentine sections during contact microradiography. We introduce a simple modification which allows the dentine to be wet during the microradiographic procedure. The volume stability of both sound and demineralized dentine sections microradiographed under wet conditions is compared to sections microradiographed while being exposed to the air. Under wet conditions no shrinkage could be detected of both sound and demineralized sections. Within 15 min exposure to the air, demineralized sections showed their lesion depth to be significantly reduced by 21%, resulting in an underestimation of the mineral loss of 44%. No shrinkage was observed in the air-dried sound dentine. Especially when high-resolution plates are used, which require an extended exposure time, microradiography of dentine sections under wet conditions is recommended. In longitudinal de- and remineralization studies, the use of water instead of impregnation with low-volatility liquids is to be preferred because of the possible effects of these liquids on the mineralization processes.

Absorptiometry, Photon↗

Major topics in quantitative microradiography of enamel and dentin: R parameter, mineral distribution visualization, and hyper-remineralization.

Three aspects of quantitative transverse microradiography are described and discussed: (1) the average mineral loss parameter of enamel or dentin lesions (R) in vol%; (2) mineral distribution visualization of lesions by means of a computer-assisted videodensitometric (CAV) method; and (3) the microradiography of hyper-remineralized lesions. "R", defined as the average amount of mineral lost (or gained) in a lesion per unit lesion length, is a useful parameter in dental caries. The results show that, in vitro, R is about constant over the demineralization period for enamel and dentin. This was found even for quite different dissolution kinetics. R is strongly reduced by the presence of small amounts of fluoride in the demineralizing system. R of dentin lesions is smaller than for enamel lesions. We conclude that the mineral loss value (delta Z) and the lesion depth (ld) are in general dependent parameters during in vitro or in situ studies on enamel or dentin. During lesion formation, mineral loss from the lesion and acid penetration in the tissue are obviously linked. This is possible only if the mineral content in the lesion does not change very much after passage of the lesion front. The CAV method presented is illustrated for enamel lesions formed in vitro and in situ.

Absorptiometry, Photon↗

Application of transverse microradiography for measurement of mineral loss by acid erosion.

This paper describes a novel application of transverse microradiography for the detection and quantification of mineral loss due to acid erosion in thin tooth sections. Sixty-four specimens were randomly divided into eight equal-sized groups and exposed to an orthophosphoric-acid-based erosive fluid (pH = 3) for 0, 0.25, 0.5, 1, 2, 5, 12, or 24 hrs. We made microdensitometric scans separately across both enamel and dentin to derive data for the total mineral loss and the minimum mineral content within the eroded area. We then analyzed specimens using a profilometer to determine (1) the area above a plot of penetration depth against distance and (2) the maximum depth of penetration. Correlation coefficients for comparisons between microradiographic and profilometric data for both enamel and dentin specimens varied between 0.87 and 0.96. Two-sample t tests demonstrated that the microradiographic technique could detect early erosion, i.e., discriminate between erosion times of less than 1 hr. It was concluded that this application of transverse microradiography was a useful and acceptable method for the measurement of early mineral loss in vitro, occurring as a result of acid erosion.

Absorptiometry, Photon↗

Comparison of backscattered scanning electron microscopy and microradiography of secondary caries.

Carious lesions are usually studied using light microscopy and/or microradiography which require preparation of thin sections. Backscattered scanning electron microscopy (BSEM) has received little attention although it provides information similar to that obtained with microradiographs, with the potential for higher resolution. Recently, microscopes have been introduced that can be used to study wet or nonconducting specimens, offering techniques for studying specimens without desiccation or preparation of thin sections. This investigation sought to determine if secondary carious lesions have the same characteristics when studied by microradiography as when using the "wet" BSEM mode. Microradiographs were made of thin sections from restored teeth with secondary caries induced in an artificial caries system. The thin sections were also studied by BSEM with a partial pressure in the specimen chamber to prevent specimen charging. Comparisons of the lesion size and shape were made using the two methods. Lesion depth measurements in enamel were the same; lesions that penetrated into dentin appeared to be of similar size and shape, but lesion depths measured by BSEM were slightly greater (paired t-test, p less than .05). This was a result of cracks at the carious enamel-dentin interface that probably developed during storage of the samples. Variations in the surface enamel rod structure and the development of subsurface lesions were apparent. Several zones were also apparent in the carious dentin, demonstrating loss of dentinal tubule detail in the depth of the lesion, collapse of tubules, and hypermineralized regions near the advancing front of the lesion. Several additional samples of natural carious teeth were examined. They demonstrated the characteristic structural features of the carious process.(ABSTRACT TRUNCATED AT 250 WORDS)

Dental Caries↗

[Contribution of microradiography in search of calcifications of valvular bioprostheses].

Implantation of a valvular bioprosthesis is one of the best treatments for valvular disease, particularly in children. Unfortunately, their use is limited as calcifications develop over time. Prevention of these calcifications is still an unresolved problem that is under study by Professor Carpentier and coworkers. These investigators have developed an experimental model in which accelerated development of calcifications occurs: fragments of porcine valves implanted under the skin of rats become calcified in a few days. Prior to implantation, the fragments are pretreated in an attempt to delay or prevent the development of calcifications. Two, four, six and eight months after implantation, the fragments are removed and calcifications are looked for using microradiography. Two hundred fragments have been studied. The material and technique used in our study are described. Results are semi-quantitative, based on the volume of calcifications. We conclude that microradiography is faster and more reliable than histology (the only technique used up till now) as it allows to study the entire fragments.

Animals↗

[[Studies of Ossification of the posterior longitudinal ligament in the cervical spine using microradiography and histochemistry (author's transl)].

ossification of the posterior longitudinal ligament (OPLL) is seen by X-ray as a condition which shows a heterotopically formed bony shadow at the posterior part of vertebral bodies of the cervical spine. OPLL causing severe myelopathy is called a Japanese disease. Recently, it has been shown that this condition is not only seen in Japan, but also in other countries especially in Asia. In order to obtain information of pathogenesis of this condition, the present author carried out histochemical investigations on 2 autopsy cases which had been treated as OPLL myelopathy, and 12 surgical materials. Totally 30 spinal vertebrae were examined. They were prepared for study with undemineralized ground sections mainly. The following results were obtained. 1) Microradiography revealed that the calcified tissue on the ligament had two marked structures, one was highly calcified tissue, and the other was bony tissue with lower density. 2) Most of the highly calcified tissue with variable distribution ranging from a sparse to a dense one contained round radiolucent spots probably consisting of cartilage-like cells. This type of calcified tissue is accompanied by the organic matrix, which was stainable with van Gieson stain, PAS reaction, Alcian blue stain and toluidin blue metachromasia. 3) A part of highly calcified tissue showed linearly distributed pattern without radiolucent round spots. This type of calcified tissue was also stained with van Gieson stain and PAS reaction. However, this organic matrix was not accompanied by marked reaction of acid mucopolysaccharides. 4) No essential histochemical difference was found between the highly calcified tissue on the ligament and other various types of heterotopically formed calcified tissue. 5) Microradiography revealed the remnant of resorption process at the boundary between the highly calcified tissue and the bony tissue. This finding of the resorption lacunae indicated that the highly calcified tissue was formed initially, followed by bone tissue formation. The process of internal remodeling was also observed in the bony tissue. 6) The distribution of the highly calcified tissue, bony tissue and resorption cavities indicated several patterns of remodeling process which was characterized by the new bone formation at the region near the vertebral body. These characteristics were apparently due to the mechanical stresses acting on this bony structures, as seen in some other heterotopic bone formations such as the healing process of fracture or tooth extraction socket.

Adult↗

Histomorphometrical analysis of the bone-implant interface: comparison of microradiography and brightfield microscopy.

Section thickness has been shown to affect the histomorphometrical measurement of bone-implant contact when analysed under brightfield microscopy. This study investigated whether microradiography of the bone-implant interface eliminated the errors associated with thick section analysis. Seven implant containing sections were utilized. Microadiographs of the thick (approximately 100 microns) sections were taken and the sections were subsequently ground to thicknesses of 50 microns and 25 microns. Photomicrographs were taken of the microradiographs and of the sections at each thickness (100, 50 and 25 microns) under brightfield microscopy. The photomicrographs were analysed for direct bone-implant contact in the cortical passage region and along the total length of the implant. The effect of section thickness on multiple fluorochrome labelling in 10 rabbit femur specimens was also examined. Centre-to-centre interlabel distance was measured for each label pair at a thickness of 100 microns and then again after the sections were ground to 50 microns and 25 microns. The thick (100 microns) sections showed a significantly greater amount of bone-implant contact than either the thin sections or the microradiographs. There was no difference in direct bone-implant contact measured by microradiography or thin sections. However, the microradiographic analysis showed a much lower variability of the bone-implant contact than the sections evaluated under brightfield microscopy. In addition, they have the added benefit of providing information on bone mineral density. Centre-to-centre interlabel distance was not significantly different for any label pair owing to section thickness. Data from this study provides evidence that the use of microradiographs for histomorphometrical analysis of the bone-implant interface is superior to brightfield analysis of thin sections owing to the lower variability of microradiographical data and the ability to obtain bone mineral density measures. Additionally, given that interlabel distance was not significantly affected by section thickness, the use of 100 microns thick sections for analysis of fluorochrome labels in cortical bone is supported.

Analysis of Variance↗

Longitudinal microradiography: a non-destructive automated quantitative method to follow mineral changes in mineralised tissue slices.

This paper presents a new powerful method, longitudinal microradiography (LMR), with which it is possible to follow the mineral content in a de- or remineralizing carious tooth sample as a function of position on the surface of the mineralizing tissue independent of the sample thickness. Planoparallel tooth slices (thickness 300-400 microns) are cut parallel to the labial tooth surface and then fixed on a PMMA holder. Repeated microradiographic images of the same tissue slice together with an aluminium step wedge are made on photographic film at different times of mineralisation. The images are scanned under a densitometer with a microcomputer and the mineral content per unit tooth area at the same discrete tooth-slice surface positions is calculated (maximum predefined area is 5 mm x 5 mm, 15 x 15 positions). The change of mineral in one tooth slice per unit tooth surface area can be calculated at different times of mineralisation locally, as well as averaged over the total tooth-slice surface area. The SD in average mineral change is 0.009 kg m-2, which corresponds approximately to the mineral loss involved with a 8 microns deep tooth enamel carious lesion. All results are stored on disc. In an example, the enamel surfaces (6 mm2) of five human tooth enamel slices were exposed to an acetic acid buffer solution (pH 4.8) containing Ca, P and F. The mineral content variations in the enamel sample were followed in time by longitudinal microradiography. Mineral content evaluation shows in all cases a gradual increase in mineral loss. 3D plots of the mineral loss per unit area against tooth surface position at different stages of demineralisation of the same sample show an excellent uniform local behaviour in time with all tooth slices.

Dental Caries↗

Microradiography as a tool to detect heavy metal uptake in plants for phytoremediation applications.

In this paper, an application of contact microradiography with soft X-rays for detecting the uptake site of heavy metal in the whole plant leaves is investigated. The X-ray source is a laser-plasma one based on an Nd:glass laser. The soft X-ray radiation emitted from the plasma laser targets of magnesium, iron, and copper can be strongly absorbed in the leaves' regions rich in iron, magnesium, and copper. This absorbance could point to structures in the leaves where these heavy elements are found. In this work, leaves treated with copper sulfate diluted in water at 1, 2, and 5% were imaged by using a copper target, in order to evaluate differences with untreated control leaves. Our results showed that this methodology highlighted the presence of copper in the treated leaves. This new methodology should detect heavy element pollutants inside plants and it should also be a useful analytic tool in phytoremediation studies.

Copper↗

Cariostatic effect of a light-cured, resin-reinforced glass-ionomer for bonding orthodontic brackets in vivo. A combined study using microradiography and confocal laser scanning microscopy.

The aim of this study was to evaluate in vivo the cariostatic potential of the resin-reinforced glass-ionomer (Vitremer core build-up restorative; 3M Dental Product Division) when used as a bonding agent for orthodontic brackets. The mineral distribution and topography of the enamel surface adjacent to the bracket base was determined by quantitative microradiography (TMR) and confocal laser scanning microscopy (CLSM). The study was designed in split-mouth technique using 9 pairs of premolars to be extracted for orthodontic reasons. One tooth of each pair was bonded with the resin-reinforced glass-ionomer, and the control contralateral premolar with the non-fluoridated composite (Concise, 3M Dental Products Division). After 4 weeks all teeth were extracted and stored until analysis. The lesion depths and mineral loss values in enamel adjacent to brackets bonded with Vitremer were significantly lower than in teeth bonded with the composite, indicating that the resin-reinforced glass-ionomers significantly reduced caries lesion development in vivo. CLSM images show a severe cariogenic challenge around orthodontic brackets and support TMR measurements.

Bicuspid↗

The degree of mineralization of bone tissue measured by computerized quantitative contact microradiography.

Strength of bones depends on bone matrix volume (BMV), bone microarchitecture, but also bone mineralization, and we have recently shown in osteoporotic patients treated with alendronate that fracture risk and bone mineral density (BMD) were changed without modifications of BMV or bone microarchitecture. Mineralization of bone matrix implies two successive steps: a rapid primary mineralization on the calcification front followed by a slow process of secondary mineralization progressively adding about one-half of the mineral content on bone matrix. These two steps are clearly illustrated by microradiographs of compact and cancellous bone tissue from transiliac human biopsies. Our working hypothesis is based on the impact of changes in bone remodeling rate on the degree of mineralization of bone, i.e., on the BMD measured at the tissue level. Contact quantitative microradiography using a computerized microdensitometric method, is described and allows the measurement of the mean degree of mineralization of bone (MDMB). This parameter may be quantitatively evaluated by exposing an aluminum calibration step-wedge and a plane-parallel calcified tissue section simultaneously to the same beam of X-rays, then determining, from the resulting microradiograph, the thickness of aluminum that produces the same X-ray absorption as a given region of the bone tissue section. To be used as a control group, iliac bone samples were taken at necropsy from 43 subjects (30 women aged 48.4 +/- 3.7 years and 13 men aged 66.0 +/- 4.4 years) who died suddenly showing no apparent bone disease. A control MDMB, which does not change with age, and a control distribution of these values are thus established. These control values are necessary for interpreting the changes in MDMB observed in bone conditions untreated or treated.

Absorptiometry, Photon↗

Microradiography and electron-microprobe analysis of some caries-like lesions of enamel prepared in vitro in human teeth.

Six lesions were made on the buccal surfaces of premolars. The volume percentage of mineral was determined as a function of depth by microradiography. Using the electron microprobe, the signals for Ca, Na, Mg, P and Cl were recorded as a function of depth both through the lesions and through the adjacent sound enamel. In the demineralized parts, there was a preferential loss of Na and a preferential retention of chloride. In the surface layers, the Na:Ca and Cl:Ca ratios were almost the same as in the adjacent sound enamel, indicating that the surface layers were not formed by gross dissolution of the original mineral followed by gross reprecipitation of another, less-soluble calcium phosphate, but remained probably because their microcrystals were protected by a thin layer of precipitated fluorapatite or fluoridated hydroxyapatite. The same had been found for surface layers in natural caries.

Calcium↗

Mineralization differences between human deciduous and permanent enamel measured by quantitative microradiography.

The mineralization levels of erupted buccal enamel from 24 deciduous teeth were compared to those of 28 permanent teeth. Sections were prepared in a defined plane using a lapping machine which gave plano-parallel sections. Mineralization levels were recorded by quantitative microradiography at 25 equivalent anatomical sites in each section. Deciduous incisors and canines were compared with their homologous successors: overall mineralization levels were lower in the deciduous dentition, with no significant differences being found close to the amelo-dentinal junction, but highly significant differences being found in the outermost sites. Deciduous molars were compared with premolars, and were also relatively less mineralized. However, deciduous molars did not show the consistent diminishing occlusocervical gradient observed in all other tooth types tested; on the contrary, they showed a cervical reversal with higher values than permanent premolar enamel. These results confirm the generally lower mineral levels in deciduous enamel, and provide quantitative information on site-specific mineralization levels.

Cuspid↗

Intra-operative specimen analysis using faxitron microradiography for excision of mammographically suspicious, non-palpable breast lesions.

Recent advances in digital imaging have made Faxitron microradiography an attractive alternative to intra-operative conventional specimen radiography (CSR) for the excision of wire-localized breast lesions. Faxitron specimen analysis time, usefulness of digital image manipulation and re-excision rates were evaluated in comparison to CSR in 299 consecutive wire-localized excisions for mammographically suspicious non-palpable breast lesions (172 procedures with Faxitron, 127 with CSR) in a non-randomized study. The corresponding mean operation times were 34.7 vs. 42.7 min and the respective re-excision rates were 19.8% vs. 31.5% (no significant difference on chi analysis P < 0.1). Faxitron digital image manipulation led to cavity biopsies in 50% (60/121) of the cancer excisions. In 19 of these (16%), histological excision margins were converted from incomplete to complete. The shorter Faxitron mean operating time enables an additional wire-localized operation per theatre list. Digital imaging guides the surgeon for additional cavity biopsies, resulting in re-excision rates as good as CSR.

Adult↗

Preliminary quantitative microradiography study into the distribution of bone mineralization within the basal bone of the human edentulous mandible.

Six edentulous human hemimandibles (three male, three female) of similar ages were sectioned transversely at the midline, mental foramen, midway between mental foramen and angle (body), and angle of the mandible. Planoparallel sections were prepared of the cortical plate on buccal and lingual surfaces and microradiographed alongside an aluminium step-wedge for computerized quantitative microradiography. Mean mineralization values and mineralization frequency distribution curves were calculated. Gender had no effect on the mean mineralization values, but did show a significant difference in the mineralization frequency distribution. There was no difference in mean mineralization between lingual and buccal cortical plates, but the distribution curves differed, with the lingual cortex distributions being more uniform. Each sample site was significantly different from all others, with the mental foramen and body sites showing the greatest variation in distribution of mineralization level.

Aged↗

Renal microradiography of experimental ureteral obstruction in the rabbit.

Microradiography of nephrons in kidneys perfusion-fixed with glutaraldehyde permits examination of large numbers of nephrons. This technique was applied to rabbits between one and 14 days following unilateral ureteral ligation. Kidneys without ureteral occlusion served as controls. By two days after ureteral obstruction there was dilatation of the ducts of Bellini and papillary collecting ducts. At three to four days there was splaying and tortuosity of the loops of Henle. By eight to 10 days the proximal straight tubules were noted to be dilated and helically twisted. After two weeks of ureteral obstruction there was dilatation of Bowman's space with encroachment on the glomerular capillary tuft. At this time many proximal convoluted tubules began to show atrophic changes. These morphologic alterations may due in part to back pressure on the nephrons, with retrograde progression as the duration of urinary tract obstruction is increased. The distal convoluted tubule and the descending limb of the loop of Henle were not noted to be abnormal during the study.

Animals↗

An in vitro assessment using transverse microradiography of the effect on mineral loss of etching enamel for in situ studies.

OBJECTIVES: To test the hypothesis that etching enamel with 37% phosphoric acid for 30 s does not lead to detectable mineral loss when measured with transverse microradiography (TMR). DESIGN: An in vitro laboratory investigation. EXPERIMENTAL VARIABLE: Forty bovine incisors were used in the experiment. The crowns of the teeth were covered with acid resistant varnish except for a rectangular area on the labial surface approximately 10 x 12 mm. On the exposed labial surface of 20 teeth an enamel lesion similar to that used in the in situ caries model was induced. Twenty teeth were left without a lesion. The exposed area was divided into three areas of equal size. The control area (C) was covered with acid resistant varnish throughout the experiment. The first experimental area (E1) was etched with 37% phosphoric acid for 30 s and a simulated bracket was bonded to the surface with composite resin. The second experimental area (E2) was left exposed for the remainder of the experiment. The teeth were placed in a demineralizing solution for 24, 48, 72 or 96 h to replicate different cariogenic challenges. OUTCOME MEASURE: Mineral loss as measured with TMR. RESULTS: There were no significant differences in the mineral loss between etched (E1) and etched (C) areas of enamel. There were significant differences in mineral loss between E1 and E2 for the 48 h (p < 0.001) and 72 h (p = 0.001) exposures without a pre-formed enamel lesion. CONCLUSION: There is no detectable mineral loss with TMR when enamel has been etched for 37% phosphoric acid for 30 s. The use of in situ enamel specimens with acid etch retained simulated brackets to investigate demineralization during orthodontics will not significantly affect the outcome compared with unetched specimens.

Acid Etching, Dental↗

Studies in histochemistry. LVII. Determination of the total dry mass of human erythrocytes by interference microscopy and x-ray microradiography.

The total dry mass of human erythrocytes was determined by both interference microscopy and x-ray microradiography. The determination of mass per unit area, and calculation of total dry mass per cell were simplified by changing the shape of the cells to spheres which were then flattened to discs of constant thickness when smeared on glass slides for measurement of fixed cells by interferometry, and to oblate spheroids when smeared on parlodion-coated slides for measurement of fixed cells by x-ray absorption. From x-ray measurements of 100 smeared and alcohol-fixed cells a mean dry mass per cell of 33.7 x 10(-12) g was obtained. Interference measurements of 100 fresh cells suspended in isotonic saline gave a mean value of 32.4 x 10(-12)g while interference measurement of 100 smeared and alcohol-fixed cells gave a mean value of 30.8 x 10(-12)g. The first two values compare well with a mean corpuscular hemoglobin of 31.2 x 10(-12)g, obtained from determinations of erythrocyte count and hemoglobin, since 95 per cent of the dry mass of the cell is hemoglobin. The difference in interference values between the fixed and fresh cells is possibly due to a difference between the specific refractive increment of alcohol-denatured hemoglobin and that of the unmodified substance. The value for the latter was used since that of the former is unknown.

Erythrocyte Indices↗