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Physical and mechanical characterization and the influence of cyclic loading on the behaviour of nickel-titanium wires employed in the manufacture of rotary endodontic instruments.

AIM: To analyse the influence of cyclic loading on the mechanical behaviour of nickel-titanium (NiTi) wires employed in the manufacture of ProFile rotary endodontic instruments. METHODOLOGY: Nickel-titanium wires, 1.2 mm in diameter, taken from the production line of ProFile rotary endodontic instruments before the final machining step, were tensile-tested to rupture in the as-received condition and after 100 load-unload cycles in the superelastic plateau (4% elongation). The wires were characterized by X-ray energy-dispersive spectroscopy, X-ray diffraction and by differential scanning calorimetry and compared with new size 30, .06 taper ProFile instruments. The fracture surfaces of the wires were observed by scanning electron microscopy. RESULTS: The mechanical properties of the as-received wires, their chemical composition, the phases present and their transformation temperatures were consistent with their final application. Only small changes, which decreased after the first few cycles, took place in the mechanical properties of the cycled wires. The stress at maximum load and the plastic strain at breakage remained the same, while the critical stress for inducing the superelastic behaviour, which is related to the restoring force of the endodontic instruments, decreased by approximately 27%. CONCLUSIONS: The mechanical behaviour of the NiTi wires was modified slightly by cyclic tensile loading in the superelastic plateau. As the changes tended towards stabilization, the clinical use of rotary NiTi ProFile instruments does not compromise their superelastic properties until they fracture by fatigue or torsional overload, or are otherwise discarded.

Calorimetry, Differential Scanning↗

Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy.

As atomic force microscopy (AFM) imaging of live specimens becomes more commonplace, at least two important questions arise: 1) do live specimens remain viable during and after AFM, and 2) is there transfer of membrane components from the cell to the AFM probe during probe-membrane interactions? We imaged live XR1 glial cells in culture by single- or dual-pass contact or tapping-mode AFM, examined cell viability at various postimaging times, and report that AFM-imaged live XR1 cells remained viable up to 48 h postimaging and that cell death rates did not increase. To determine if nonlethal, transient interactions between the AFM probe and cell membrane led to transfer of XR1 cell membrane phospholipid components on the probe, we treated the scanned probes with the lipid-binding fluorophore FM 1-43. Confocal microscopy revealed that phospholipid membrane components did accumulate on the probe, and to a generally greater extent during contact-mode imaging than during tapping-mode imaging. Moreover, membrane accumulations on the probe were greater when live XR1 cells were damaged or perturbed, yet membrane did not accumulate in fluorescently detectable quantities during repeated "force curves" during control experiments. Taken together, our data indicate that although AFM imaging of live cells in culture does not affect long-term cell viability, there are substantial probe-membrane interactions that lead to transfer of membrane components to the probe.

Animals↗

Blood and cell compatibility of gelatin-carrageenan mixtures cross-linked by glutaraldehyde.

Mixtures of gelatin and iota-carrageenan cross-linked by glutaraldehyde were prepared and their physical properties and blood and cell compatibility were compared to gelatin as a control material. According to scanning electron microscopic observation of fracture surfaces, the mixtures were composed of dispersed and continuous domains which might be generated by phase separation of carrageenan. The thermal degradation temperature of iota-carrageenan in the mixtures rose with increasing gelatin content. The swelling process in the mixtures proceeded slower than in gelatin. Tensile strengths of the mixtures, except that containing 50% iota-carrageenan, increased with increased amounts of iota-carrageenan in the mixtures. The iota-carrageenan contents at the surfaces of the mixtures were generally higher than those admixed originally. Static friction coefficients of the mixtures were lower than that of gelatin. Plasma recalcification times of the mixtures were longer than that of gelatin. Platelet adhesion of the mixtures was lower than that of gelatin, while cell adhesion and growth assays using Chinese hamster ovary cells showed that cell adhesion and growth were not dependent on adding iota-carrageenan. It was concluded that blood compatibility of the mixtures increased and cell compatibility did not decrease, compared to gelatin.

Animals↗

The application of physico-chemical procedures in the analysis of urinary calculi.

All physico-chemical techniques used in the analysis of urinary calculi have inherent advantages and limitations. Although x-ray powder diffraction can identify constituents unambiguously, certain minor components can be missed. Infrared spectroscopy is more sensitive but band assignment at low concentrations is difficult. Scanning electron microscopy together with energy dispersive x-ray analysis permits the simultaneous investigation of morphology and chemical microstructure. However, microanalysis of elements lighter than sodium is not possible and constituents are prone to irradiation damage. With the electron microprobe, minor constituents can be detected but tedious sample preparation procedures are required. Transmission electron microscopy is extremely useful in determining constituent inter-relationships and ultrastructure but ultramicrotomy is very difficult. Thermal gravimetric analysis gives quantitative information easily but does not satisfactorily distinguish between struvite and brushite. In an attempt to assess the accuracy of chemical analyses, 62 calculi were investigated applying several chemical tests. Those for Mg2+, PO4(3-), NH4+ and uric acid proved highly reliable while that for Ca2+ often yielded an incorrect result. The test for oxalate was totally unsatisfactory. Investigators of stone composition and structure should include x-ray diffraction (or infrared spectroscopy) and scanning electron microscopy as their methods of first choice. In addition, chemical or thermogravimetric analyses should be utilized in an auxiliary capacity.

Chemistry, Physical↗

Backscattered electron imaging for the life sciences: introduction and index to applications - 1961 to 1980.

Backscattered electron imaging (BEI) in a scanning electron microscope has been used in biological investigation since 1961. Originally used for imaging specimen surfaces during x-ray microanalysis, applications of this method accelerated rapidly with the recognition of its potential for selective imaging of heavy metal stained biological structures, including those labelled by cytochemical methods. This use of backscattered imaging, to view heavy metal stained structures in cells and tissues, has become the predominate BEI application in the life sciences. This paper presents a brief introduction to BEI as it relates to the use of the scanning microscope for histo - and cytochemistry. Biological applications reported in the years 1961-1980 are cited and indexed by subject and author(s).

Animals↗

Distribution of concanavalin A binding sites on normal human urinary bladder mucosa and bladder tumors by transmission and scanning electron microscopy and X-ray microanalysis.

We used concanavalin A (con A)-peroxidase-iron dextran-diaminobenzidine (DAB) technique for the electron microscopic detection of con A binding sites on cell membranes. Normal bladder mucosa showed a sparse distribution of con A binding sites with both transmission (TEM) and scanning (SEM) electron microscopy, but bladder tumors showed a higher concentration in the distribution of con A binding sites in proportion to the histopathological grade of transitional cell carcinoma. Quantitative estimation of the con A binding sites was attempted using scanning X-ray pulse analysis of iron elements contained in the reaction complexes. Con A binding sites were quantitatively the smallest in normal mucosa, increasing proportionate to the grade of the bladder tumor. Some specimens were compared by the ferritin-labelled method and the pattern of ferritin conjugates distribution was similar to that seen with the con A-peroxidase-iron dextran method.

Carcinoma, Transitional Cell↗

Hydroxyapatite-based porous aggregates: physico-chemical nature, structure, texture and architecture.

At the request of medical teams from the maxillofacial sector, a highly porous ceramic support based on hydroxyapatite of around 70-80% porosity was produced with a pore size distribution similar to bone texture (< 10 microns, approximately 3 vol%; 10-150 microns, approximately 110 vol%; > 150 microns, approximately 86 vol%). The ceramic substrates were conceived not only as a fillers for bone cavities, but also for use as drug dispensers and as supports to host cells to produce particular therapeutic agents. A method is suggested to obtain a substrate of high porosity, exploiting the impregnation of spongy substrate with hydroxyapatite ceramic particles. X-ray and scanning electron microscopy analyses were carried out to evaluate the nature of the new ceramic support in comparison with the most common commercial product; pore size distribution and porosity were controlled to known hydroxyapatite ceramic architecture for the different possible uses.

Biocompatible Materials↗

Scanning Single-Electron Transistor Microscopy: Imaging Individual Charges

A single-electron transistor scanning electrometer (SETSE)-a scanned probe microscope capable of mapping static electric fields and charges with 100-nanometer spatial resolution and a charge sensitivity of a small fraction of an electron-has been developed. The active sensing element of the SETSE, a single-electron transistor fabricated at the end of a sharp glass tip, is scanned in close proximity across the sample surface. Images of the surface electric fields of a GaAs/AlxGa1-xAs heterostructure sample show individual photo-ionized charge sites and fluctuations in the dopant and surface-charge distribution on a length scale of 100 nanometers. The SETSE has been used to image and measure depleted regions, local capacitance, band bending, and contact potentials at submicrometer length scales on the surface of this semiconductor sample.

Journal Article↗

[Relation between changes in platelet shape, calcium balance and the separating effect of cytochalasin B].

Platelets were stimulated in vitro by different ADP concentration (10(-7)--10(-9) M). Scanning electron microscopy, light transmission recording and the Ca-sensitive fluorescent probe, chlortetracycline, were used to study the time course of changes in the shape, aggregation and the amount of membrane-bound calcium during ADP activation. It was found that ADP (10(-7)--10(-8)M) causes reversible changes in the shape of platelets without disturbing their aggregation. The changes in question were accompanied by phasic alterations of the amount of membrane-bound calcium.

Adenosine Diphosphate↗

[Effect of new photosensitizer CDHS801-mediated photodynamic therapy on bladder cancer: an experimental study].

OBJECTIVE: To investigate the effects of dynamic photodynamic therapy (PDT) on bladder cancer. METHODS: Human bladder cancer cells of the line T24 were co-cultured with CDHS801, a photosensitizer, and MitoTracker RED CMXRose and MitoTracker GREEN FM, mitochondria specific fluorescence probe dyes. Laser scanning confocal fluorescence microimaging system was applied to collect the fluorescence of the photosensitizer and the probes. T24 cells were cultured and divided into 4 groups: Group 1 as blank control group, Group 2 undergoing laser irradiation, Group 3, added with CDHS801 for 6h, and Group 4 (PDT group), added with CDHS801 and undergoing laser irradiation. The survival of the cells was examined by MTT colorimetric assay. The morphological changes and apoptosis of the photo-activated T24 cells were investigated by transmission electron microscopy, confocal laser scan microscopy, and flow cytometry. RESULTS: The fluorescence of the photosensitizer and that of the probes were detected in the cytoplasm and in the peri-nuclear region, mainly in the mitochondria, of the T24 cells. 2. The inhibitory rates of PDT on T24 cells were 0%, 7.3%, 10.8%, and 71.4% in the control group, Group 1, Group 2, and Group 3 respectively. T24 cell photo-activated with CDHS801 showed cell size shrinkage, condensed chromatin and formation of apoptotic body. Flow cytometry showed that apoptosis was seen in 55.31% of the photo-activated cells and peaked in the sub-G1 phase. However, no such changes were seen in the control group. CONCLUSION: CDHS801-based PDT can kill bladder cancer T24 cells. CDHS801 is localized in the cytoplasm and peri-nuclear region, mainly mitochondria, of tumor cell. CDHS801 based PDT maybe eliminate the T24 cell by the induction of apoptosis.

Apoptosis↗

Numerical dominance and phylotype diversity of marine Rhodobacter species during early colonization of submerged surfaces in coastal marine waters as determined by 16S ribosomal DNA sequence analysis and fluorescence in situ hybridization.

Early stages of surface colonization in coastal marine waters appear to be dominated by the marine Rhodobacter group of the alpha subdivision of the division Proteobacteria (alpha-Proteobacteria). However, the quantitative contribution of this group to primary surface colonization has not been determined. In this study, glass microscope slides were incubated in a salt marsh tidal creek for 3 or 6 days. Colonizing bacteria on the slides were examined by fluorescence in situ hybridization by employing DNA probes targeting 16S or 23S rRNA to identify specific phylogenetic groups. Confocal laser scanning microscopy was then used to quantify and track the dynamics of bacterial primary colonists during the early stages of surface colonization and growth. More than 60% of the surface-colonizing bacteria detectable by fluorescence staining (Yo-Pro-1) could also be detected with the Bacteria domain probe EUB338. Archaea were not detected on the surfaces and did not appear to participate in surface colonization. Of the three subdivisions of the Proteobacteria examined, the alpha-Proteobacteria were the most abundant surface-colonizing organisms. More than 28% of the total bacterial cells and more than 40% of the cells detected by EUB338 on the surfaces were affiliated with the marine Rhodobacter group. Bacterial abundance increased significantly on the surfaces during short-term incubation, mainly due to the growth of the marine Rhodobacter group organisms. These results demonstrated the quantitative importance of the marine Rhodobacter group in colonization of surfaces in salt marsh waters and confirmed that at least during the early stages of colonization, this group dominated the surface-colonizing bacterial assemblage.

Colony Count, Microbial↗

Bonding of chemically treated titanium implants to bone.

A study was undertaken in rabbit tibiae to determine the effects of chemical treatments and/or surface-induced bonelike apatite on the bone-bonding ability of titanium (Ti) implants. Smooth-surfaced plates (10 x 10 x 2 mm) of pure Ti, alkalil- and heat-treated Ti, and bonelike apatite-formed Ti after the treatments were implanted into the tibial metaphyses of mature rabbits. The tibiae containing the implants were harvested at 4, 8, and 16 weeks after implantation and subjected to a tensile testing and histologic evaluation. Biomechanical results showed that both treated implants exhibited significantly higher failure loads compared with untreated Ti implants at all time periods. Histologic examination by Giemsa surface staining, contact microradiography (CMR), and scanning electron microscopy (SEM) in backscatter mode revealed that both treated Ti implants directly bonded to bone tissue during the early postimplantation period, whereas untreated Ti implants formed direct contact with the bone only at 16 weeks. SEM-electron-probe microanalysis (EPMA) examination showed a Ca-P-rich layer at the interface between the treated implants and bone, although the Ca-P-rich layer was not detected on the surface of untreated implants during observation periods. The results of this study suggest that chemical treatments may accelerate the bone-bonding behavior of titanium implants and enhance the strength of bone-implant bonding by inducing a bioactive surface layer on Ti implants.

Animals↗

Study of cystine urinary calculi in dogs.

The composition and structure of 48 canine cystine urinary stones were determined by infrared spectroscopy, scanning electron microscopy and electron dispersive X-ray analysis. The infrared analysis showed that about 45% of the specimens were composed of pure cystine. The remainder also contained calcium oxalate (mono and/or dihydrate), magnesium ammonium phosphate hexadydrate (struvite), calcium hydrogen phosphate dihydrate (brushite) and complex urates (ammonium, ammonium potassium and/or potassium enriched ammonium urate). The infrared study of several samples heated at 620 degrees C and 750 degrees C revealed the presence of apatitic calcium phosphate. This compound was difficult to detect in the spectrum of the original samples due to the small proportion of phosphate contained in the calculi and to band overlapping. The examination of a series of selected samples by means of scanning electron microscopy and energy dispersive X-ray analysis complemented the infrared results.

Animals↗

Aortic endothelium of alloxan diabetic rabbits: a quantitative study using scanning electron microscopy.

Scanning electron microscope studies of the aorta and other major arteries have been performed in alloxan-induced diabetic rabbits. After 5 weeks, a variety of structural abnormalities of the endothelial lining were detected including a significant increase in the number of argyrophilic cells and an increased number of craters or openings in the endothelial junctional region. Evidence of more extensive micro-damage was present after 5 months duration of diabetes. These zones with structural changes in the endothelial lining of major vessels seem to be areas of high predilection to atherosclerosis in diabetes.

Animals↗

Physical and chemical characterization of mn phosphate/sulfate mixture used in an inhalation toxicology study.

The use of methylcyclopentadienyl manganese tricarbonyl (MMT) in unleaded gasoline has given rise to numerous debates on the potential public health risk associated with manganese emissions. In fact, combustion products are mainly Mn phosphate, Mn sulfate, and Mn phosphate/sulfate mixture. Our research group did several inhalation studies in order to assess the toxicity of each Mn species. The objective of this study is to determine the physical and the chemical characteristics of a mixture of Mn phosphate/sulfate used in one of these inhalation toxicology studies. First, the mixture was analyzed by X-ray diffraction in order to obtain the specific peak of Mn phosphate and Mn sulfate. These peaks were used as reference. Second, samples of the mixture were collected on filters in the inhalation chamber at a concentration level of 3000 microg/m(3). They were analyzed by scanning electron microscopy (SEM), analytical transmission electron microscopy (ATEM), and x-ray energy-dispersive spectrometry (EDS) to show their size, morphology, and chemical composition. Results indicate that 33% of the particles were found to be agglomerated, while free particles accounted for 44% for Mn phosphate and 23% for Mn sulfate.

Electron Probe Microanalysis↗