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Three-dimensional ultrasound biomicroscopy, environmental and conventional scanning electron microscopy investigations of the human zonula ciliaris for numerical modelling of accommodation.

PURPOSE: Biomechanical modelling of the accommodation process is a useful tool for studying the mechanism of accommodation and presbyopia and can aid in the development of accommodative lens-replacing materials. Existing biomechanical models, however, use a very simplified zonula structure. The aim of this study was to use three-dimensional ultrasonic imaging and scanning electron microscopy to provide a more detailed, three-dimensional description of the structure of the human zonula to improve the modelling of accommodation. METHODS: Five human eyes were examined without invasive manipulation using a custom-made three-dimensional ultrasonic imaging technique that allows scanning of features with a spatial resolution of 30 microm. Environmental and conventional scanning electron microscopy (SEM) provided information to complement the ultrasonic images for use in development of a more anatomically correct finite-element model of the zonula structures. These data along with the material properties of the ocular tissue structures were used to construct an advanced geometric model for finite-element simulation of the accommodation process. RESULTS: Images were obtained through three-dimensional ultrabiomicroscopy (3D-UBM) of anatomical features heretofore not directly imagable in their native state. Ciliary processes and zonula structures were clearly separated by both the 3D-UBM and the SEM methods. It was found that fibres inserting on the anterior and posterior lens capsule emerge anteriorly at the ciliary body. Fibres emerging near the pars plana insert on the lens and the ciliary body. No X-shaped crossing fibres were found. Modelling of the accommodation process with both the simple and the more complex geometric models produced refractive power changes comparable with in vivo findings. CONCLUSIONS: The 3D-UBM allowed examination of zonula structures in their native state with minimized preparation artefacts. While these data were incorporated into a complex and more anatomically correct finite-element simulation of intraocular features including lens, zonular system and ciliary body it was found that a simplified zonular model is sufficient for the numerical simulation of the accommodation process.

Accommodation, Ocular↗

Air abrasion of failed bonded metal brackets: a study of shear bond strength and surface characteristics as determined by scanning electron microscopy.

Failed bonded orthodontic brackets generally require their replacement with a new bracket. The introduction of air abrasion (microetching) technology to orthodontics may allow for immediate use of these failed brackets. This study compared the in vitro shear bond strengths of previously failed bonded metal brackets (experimental group) subjected to air abrasion with new untreated brackets (control group). All bonding used a light-cured orthodontic bonding system. In addition, representative samples from each group were examined by scanning electron microscopy. The results of this study found no significant differences in shear bond strengths between the two groups. Scanning electron microscopy examination of the air abraded brackets revealed a roughened mesh surface with residual bonded material in the bracket base. This simple technique should allow for the immediate reuse of previously failed bonded metal brackets.

Acid Etching, Dental↗

Apical tubular network in the rat kidney proximal tubule cells studied by thick-section and scanning electron microscopy.

The apical cytoplasm of several absorbing epithelia contains well-developed apical tubules (AT) which contribute to membrane recycling from endocytic vacuoles to the apical cell membrane. In this study, we examined three-dimensional structures of the AT in rat kidney proximal tubule cells by transmission and scanning electron microscopy. In thin sections, the AT appeared as straight tubules with a rather constant diameter (70-90 nm), but others were curved and, occasionally, branching. No AT were labeled with the marker for the external cell surface (ruthenium red) or exhibited histochemical enzyme activity for lysosomal hydrolase (acid phosphatase). After intravenous injection of horseradish peroxidase, it was absorbed in the kidney proximal tubule cells and the AT were labeled with HRP reaction products. Stereo-viewing of the labeled AT in thick sections revealed that they formed an interconnected tubular network. Scanning electron microscopy allowed a three-dimensional view of the AT, in which a network of branching and anastomosing tubules was revealed. These observations indicate that the AT are intracellular endosomal compartments which form an extensive tubular network in the apical cytoplasm. The possibility that this apical tubular network serves as a large membrane store for membrane recycling is discussed.

Acid Phosphatase↗

The ultrastructure of the lung after exposure to ionizing radiation as seen by transmission and scanning electron microscopy.

Male mice of the BALB/c strain were exposed at an age of 12-14 weeks, to different doses of X-rays, either to the entire thorax or to the right hemithorax. At various times after exposure mice were sacrificed and the lung was examined by transmission or scanning electron microscopy. Radiation symptoms following exposure to 15-20 Gy can be divided into three phases: Early, from a few hours to a few weeks after exposure, changes in capillary permeability and morphological alterations in all types of lung cells are prominent. At intermediate times, from a few weeks to about 7 months after exposure, the symptoms of radiation pneumonitis arise essentially, as a consequence of the action of radiation on the epithelium of the alveoli, and are characterized by a large increase in size, and probably in number of type II epithelial cells. Scanning electron microscopy brings additional arguments that the lesions in the epithelial cells, mainly of type II, are the principal cause of radiation pneumonitis and that capillary damage probably plays a secondary role.

Animals↗

Campylobacter pyloridis in peptic ulcer disease: microbiology, pathology, and scanning electron microscopy.

After the recent successful isolation of spiral organisms from the stomach this paper presents the bacteriological and pathological correlation of gastric antral biopsies from 51 patients endoscopied for upper gastrointestinal symptoms. Campylobacter pyloridis was cultured from 29 patients and seen by either silver staining of the biopsy or scanning electron microscopy in an additional three. The organism was cultured from 23 of the 33 (69%) patients with peptic ulcer disease and from within this group 17 (80%) of the 21 patients with duodenal ulceration. It was cultured only once from the 12 normal biopsies in the series but from 27 of the 38 (71%) biopsies showing gastritis. C pyloridis was also cultured from five out of seven of the 14 endoscopically normal patients, who despite this had biopsy evidence of gastritis. It was the sole organism cultured from 65% of the positive biopsies and scanning electron microscopy invariably revealed it deep to the surface mucus layer. C pyloridis persisted in the three patients with duodenal ulcers after treatment and healing. The findings support the hypothesis that C pyloridis is aetiologically related to gastritis and peptic ulceration though its precise role still remains to be defined.

Campylobacter↗

Investigation of surface changes in different types of ventilation tubes using scanning electron microscopy and correlation of findings with clinical follow-up.

OBJECTIVES: Like all biomaterials, ventilation tubes are subject to formation of bacterial biofilm on their surfaces. There might also be surface changes. This increases the risk of complications associated with ventilation tubes. In this study, we examined two groups of ventilation tubes using scanning electron microscopy (SEM) to investigate biofilm growth and surface deformations. METHODS: There were two different types of ventilation tubes used, the first group consisted of 30 silicone tubes and the second group consisted of 16 ionized, processed silicone tubes. The tube samples included those that were either removed or those that were extruded into the ear canal. We investigated the association between scanning electron microscopy findings and the complications that developed during the treatment. RESULTS: As a result of this study, it is found that the ionized, processed silicone tubes are superior to other silicon ventilation tubes in regard to biofilm growth (z=-3.925, p=0.000, <0.0001) and surface deformations (chi(2)=9.120, p=0.003, <0.01). Furthermore, we observed that as the duration of the ventilation tube application increases, bacterial biofilm growth (chi(2)=10.718, p=0.005, <0.01) and surface deformations (z=-2.940, p=0.003, <0.01) increase. We also observed that biofilm growth and occurrence frequency were related to "otorrhea" (chi(2)=10.258, p=0.036, <0.05) and "plugging" (chi(2)=7.952, p=0.019, <0.05) complications. CONCLUSION: In this study, we show that ionized, processed silicone ventilation tubes are more robust to bacterial biofilm growth compared to other silicone ventilation tubes used in this study and that the "otorrhea" and "plugging" complications are reduced with the decrease of bacterial biofilm growth.

Biocompatible Materials↗

Some "sealants" seal--a scanning electron microscopy (SEM) investigation.

White spot lesions of enamel around orthodontic brackets as a result of demineralization have been well documented in the orthodontic literature. Various methods of treatment have been attempted to reduce or eliminate this danger. The purpose of this study was to evaluate, by means of scanning electron microscopy, the polymerization of the sealant layer around orthodontic brackets with direct and indirect methods of bonding. Twenty-four sound human lateral maxillary incisor teeth were collected, cleaned, divided equally into four groups A through D, and stored in 70% ethyl alcohol. Their buccal surfaces were pumiced, etched with 37% phosphoric acid for 1 minute, and washed under running water for 30 seconds. Metal brackets were bonded with the chemically cured BIS-GMA resin, Ortho Concise, as follows: group A, indirectly bonded with coping; group B, indirectly bonded without coping; and group C, directly bonded; light activated Transbond was used in group D, directly bonded brackets. After washing in alcohol for 20 seconds, all teeth were dried, and sectioned longitudinally, through the middle of the bracket. All were subjected to 5% hydrochloric acid for 30 seconds and then washed under running water for 30 seconds. After drying and sputter coating, the teeth were viewed under scanning electron microscopy. Groups A and D showed a sealant layer surrounding the brackets and covering the buccal enamel. Groups B and C showed total absence of a cured sealant layer around the brackets or surrounding enamel.(ABSTRACT TRUNCATED AT 250 WORDS)

Bisphenol A-Glycidyl Methacrylate↗

Fetal circulation of the placenta: a comparative study of human and baboon placenta by scanning electron microscopy of vascular casts.

This study was undertaken to compare the ultrastructural appearance of the fetal microvasculature of the baboon and human placenta by the latex injection corrosion-cast technique. The general pattern of distribution of blood vessels in the placenta and the microvasculature were examined primarily by scanning electron microscopy using single and double latex (Cementex) injections of the placentae. The results of these latex injections of the baboon placenta demonstrate that (1) all the fetal arterial vessels of the baboon placenta at about three-quarters of the gestation period have a characteristic tortuosity while the venous vessels have a straight appearance, with no distinct coiling, (2) the arterial vessels exhibit a rough surface contour, (3) on the venous side the wide-calibre vessels are fairly straight and exhibit a smooth luminal surface contour, and (4) the capillary network on the arterial side is dense and highly convoluted, while on the venous side the capillary network is less tortuous. Moreover, no arterial-venous anastomosis was observed by scanning electron microscopy (SEM). The main findings based on observations of the SEM appearance of the human placental casts at three-quarter gestation and at term include the following: (1) the arterial vessels do not exhibit a marked tortuosity but do retain a rough surface contour, (2) stem vessels appear to branch at a horizontal plane before dividing into numerous smaller branches, (3) the venous side has straight, wide-calibre vessels with smooth surface contour, and (4) the capillary bed is densely tortuous on the arterial side and less tortuous on the venous side.

Animals↗

Different patterns of collagen-proteoglycan interaction: a scanning electron microscopy and atomic force microscopy study.

The extracellular matrix of unfixed, unstained rat corneal stroma, visualized with high-resolution scanning electron microscopy and atomic force microscopy after minimal preliminary treatment, appears composed of straight, parallel, uniform collagen fibrils regularly spaced by a three-dimensional, irregular network of thin, delicate proteoglycan filaments. Rat tail tendon, observed under identical conditions, appears instead made of heterogeneous, closely packed fibrils interwoven with orthogonal proteoglycan filaments. Pre-treatment with cupromeronic blue just thickens the filaments without affecting their spatial layout. Digestion with chondroitinase ABC rids the tendon matrix of all its interconnecting filaments while the corneal stroma architecture remains virtually unaffected, its fibrils always being separated by an evident interfibrillar spacing which is never observed in tendon. Our observations indicate that matrix proteoglycans are responsible for both the highly regular interfibrillar spacing which is distinctive of corneal stroma, and the strong interfibrillar binding observed in tendon. These opposite interaction patterns appear to be distinctive of different proteoglycan species. The molecular details of proteoglycan interactions are still incompletely understood and are the subject of ongoing research.

Animals↗

[New results in the visualization of the spinal dura mater with scanning electron microscopy].

UNLABELLED: Although there are various published descriptions of the dura mater spinalis [4, 7, 9, 11, 16], some points relating to the texture of the collagen fibres in the dura have still not been adequately explained. In this study the orientation of the collagen fibril bundles was revealed with the aid of scanning electron microscopy, and our observations have yielded new insights into the three-dimensional structure of the human dura mater spinalis. MATERIALS AND METHODS: The preparations used were taken from the bodies of four persons who had died of acute cardiac infarct at the ages of 70-78 years. The histories of these patients gave no indications of earlier neurological, endocrine or septic illnesses. The tissue examined was taken 8-12 h after death in all cases; it was immediately fixed in glutaraldehyde and then processed for scanning electron microscopy in the usual way. RESULTS: In the outermost (epidural) layer of the dura mater spinalis the collagen fibres are bunched together in bands that run in all directions. Elastic fibres 2 mm thick are woven into this three-dimensional network of collagen systems. On the inside (the arachnoid side) thin collagen fibres are fused into layers in such a way that the innermost layer resting on the arachnoid has a smooth, shiny appearance comparable to that of a serosa. It is attached to the actual dura with a supporting band of connective tissue. Rests of the subdural neuroepithelium could contribute to the smooth appearance of the superficial aspect. CONCLUSIONS: The outermost layer of the dura is made up mainly of collagen fibres, which run in all three directions--longitudinal, horizontal and transverse--both singly and in groups. These findings are at odds with "classic" descriptions, according to which the fibres in the dura mater spinalis all have a parallel course with a longitudinal orientation in tangential sections.

Aged↗

Conjunctival tissue examination in severe eye burns: a study with scanning electron microscopy and energy-dispersive X-ray analysis.

BACKGROUND: Long-lasting inflammation is a major problem in treatment after severe eye burns and may find expression in an altered elemental composition of the conjunctiva. Particulate contamination of biological tissue induces such inflammatory processes. In the anterior eye segment, trauma or subsequent therapy may give rise to such contamination. Scanning electron microscopy and energy-dispersive X-ray analysis are able to detect traumatic residues of submicron size and changes of the elemental composition. METHODS: Conjunctival specimens from first-time peridectomy of three healthy and nine severely burnt-eyes were examined with scanning electron microscopy and energy-dispersive X-ray analysis. The samples were prepared as cryo- or paraffin sections, mounted on carbon blocks and coated with evaporated elemental carbon. RESULTS: The samples of healthy conjunctiva showed higher concentrations of Na, P and CI. These elements showed lower concentrations in conjunctival stroma of burnt eyes excised before the 20th day after trauma than in material obtained subsequently. In two burnt conjunctival specimens there was severe traumatic contamination with Ca in Ca(OH)2 and CaO burns, and in one case the traumatic substance was Si, in a peroxide plus silicone spray burn. In the remaining six cases, particulate contamination with Fe, Al, Ni, Zn, Cu, Ti and other substances was present in the burnt conjunctivas, while no contamination was detected in the specimens of healthy conjunctivas. CONCLUSIONS: The origin of the contaminant particles is assumed to be the trauma itself and the subsequent therapy. These investigations stress the importance, for clinical purposes, or early peridectomy and contamination-free therapy.

Burns, Chemical↗

Scanning electron microscopy of black stain on human permanent teeth.

Black stain may develop on the coronal surfaces of human teeth, and this type of stain is common in the Hong Kong Chinese population. The present study was undertaken to ascertain if the deposit conforms in composition to the black stain found elsewhere, and to describe its ultrastructure using the scanning electron microscope. Gram-stained smears were made from black stain on the teeth of 11 adult Hong Kong Chinese and studied microscopically. From another 15 persons extracted teeth exhibiting black stain were obtained and fixed. Two ground sections were made from each tooth, one was stained with toluidine blue, the other was dried and prepared for scanning electron microscopy. The gram-stained smears demonstrated predominantly gram-positive filamentous microorganisms with an admixture of gram-positive cocci and rods. The ground sections revealed a deposit on the outer surface of the enamel, which was clearly divided into two distinctly different layers: an inner yellow opaque layer, and an outer layer of microorganisms. Scanning electron microscopy demonstrated that the deposit consisted entirely of microorganisms, and that in portions close to the enamel they were often obscured by a substance indicative of calcification. Thus the black stain found on the teeth of Hong Kong Chinese is similar in composition and structure to that reported to occur in other populations. The black stain is a special type of dental plaque characterized by its simple flora and its tendency to calcify.

Adult↗

Cause of spontaneous echocardiographic contrast as assessed by scanning electron microscopy.

Spontaneous echo contrast ("echocardiographic smoke") is known to occur in low-flow states and to require the presence of red cells and plasma proteins. Limited morphologic information regarding the microanatomic structure of red cells exhibiting spontaneous echo contrast is known. The study was designed to evaluate the microanatomic features of red cells exhibiting spontaneous echocardiographic contrast with scanning electron microscopy. With human blood, a beaker, and a stirring bar, a simple model for demonstration of spontaneous echo contrast and its reversal was devised. Blood elements were "sampled" from within this model at times of high and low spontaneous echogenicity by adherence of blood elements to poly-L-lysine-coated slides that were subsequently fixed and examined with scanning electron microscopy. Spontaneous echo contrast was maximal at complete stasis or low-flow states and could be abolished by agitation of blood with continuous stirring. Sampling during low-flow states with high echogenicity showed a preponderance of clumped red cells, whereas at high-flow and low echogenicity states red cells were dispersed and usually solitary. No morphologic features suggestive of activation of the coagulation system were noted. Spontaneous echo contrast is caused by reversible red blood cell clumping, which occurs in fresh human blood at low-flow (low shear rate) states and can be abolished by increasing flow. This phenomenon is independent of activation of the clotting system.

Blood Specimen Collection↗

Sealing root canals with low-viscosity resins in vitro: a scanning electron microscopy study of canal cleansing and resin adaption.

Low-viscosity resins of the fissure sealant type have been suggested in the literature as having potential for use as root canal filling materials. A low-viscosity resin may seal a root canal by flowing into clean dentinal tubules after smear layer removal. This investigation with scanning electron microscopy examines the efficacy of two methods of root canal preparation and the effectiveness of different chemicals on smear layer removal. Ultrasonic preparation with 0.25% sodium hypochlorite solution and final agitation with 50% citric acid solution were found to produce a very clean canal wall, free of smear layer in coronal and middle parts. However, low-viscosity resin used in conditions that aimed to simulate in vivo conditions failed to penetrate open dentinal tubules to a significant extent. On the basis of these observations made with scanning electron microscopy, low-viscosity resins would not seem suitable as root canal filling materials, because they are unlikely to form a satisfactory adaption to the canal wall. In addition, if treatment fails, these resins are impossible to remove from a root canal without much destruction of tooth substance.

Citrates↗

Morphometry, scanning electron microscopy and X-ray spectral microanalysis of protostylid pits on human lower third molars.

Protostylid pits on 50 lower third molars were studied. The depth and width of the pits, the thickness of the enamel at the pit bottom and the concavity of the dentino-enamel junction under the pit were measured by light microscopy. The pit content was examined by scanning electron microscopy and X-ray spectral microanalysis. The results show that the morphometric variables of protostylid pits are similar to those of occlusal fissures and pits. The protostylid pits were seen to contain globular formations, belonging mainly to dental calculus; organic rod-shaped bodies, which could represent remnants of the enamel organ--or more probably--foreign bodies, were frequently observed. The study suggests that the development of the most common form of protostylid is basically similar to that of an occlusal cusp, with variations attributable to the low expression of the protostylid and particularities of cusp development in the lateral tooth wall.

Calcium↗

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↗

Starch granule surface imaging using low-voltage scanning electron microscopy and atomic force microscopy.

High resolution imaging of wheat and potato starch granule surfaces has been performed using low-voltage scanning electron microscopy and atomic force microscopy. The complimentary images of uncoated granules demonstrate that the two starch types possess substantially different surface topologies; potato starch has many protrusions (100-300 nm in diameter), above a flatter surface containing 20-50 nm structures, whilst wheat starch possesses far fewer protrusions and generally has a smoother surface composed of approximately 20 nm structures. The protrusions are believed to be carbohydrate in nature and thus represent the ends of amylopectin side-chain clusters at the granule surface.

Microscopy, Atomic Force↗

The role of scanning electron microscopy in the diagnosis of human laryngeal cancer.

Malignant and non-malignant human laryngeal tissue samples (31 cases) were examined first by light microscopy (LM) and then the specimens were reprocessed for scanning electron microscopy (SEM). The presence of LM and SEM signs of malignancy were compared with each other. In all the 22 histologically unambiguous cases the LM and SEM results were in accordance with each other. From the 9 histologically doubtful cases, 2 were found malignant and 3 benign by retrospective SEM. The case histories of these 5 patients have confirmed the SEM results. This preliminary study suggests the application of SEM in histologically doubtful laryngeal biopsies.

Carcinoma, Squamous Cell↗