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The objective of this study was to examine the osteoinductive capacity of different concentrations of BMP-2 on bone marrow stromal cells in vitro. Further, we intended to determine whether titanium provided with an increased surface roughness is more efficient in osteoblast differentiation than machined titanium. Therefore, 20,000 cells/ml were seeded and cultured on machined and grit-blasted titanium discs for 4, 8 and 16 days. Different concentrations of rhBMP-2 (0, 10, 100, 1000 ng/ml) were supplemented to the medium for 8 days of culturing. To evaluate cellular proliferation and differentiation, specimens were examined for DNA, alkaline phosphatase activity, and calcium content. Morphological appearance of the specimens at 8 and 16 days of incubation was evaluated using scanning electron microscopy. Two separate experimental runs were performed. Evaluation of the DNA and alkaline phosphatase data revealed that a significant difference existed for these data between both experimental runs. Further analysis of the DNA figures learned that roughening of the titanium surface and addition of BMP-2 had no effect on cell proliferation. The alkaline phosphatase analysis and calcium measurements revealed that BMP-2 stimulated the early differentiation of osteogenic cells on machined titanium substrates in a dose-dependent manner. After 16 days of culture, no significant differences in calcium content could be observed anymore between machined and roughened titanium surfaces. Further, the data revealed that the machined surfaces showed a significant increase in calcium deposition when 100 and 1000 ng/ml BMP-2 were supplemented to the medium. However, the roughened surfaces showed this significant enhancement in calcium content only with 1000 ng/ml BMP-2. In addition, SEM evaluation revealed a dose-dependent response to BMP-2. Increasing BMP-2 concentrations resulted in more calcified globular accretions on bone surfaces than when no BMP-2 was added. On the basis of our results, we conclude that (1) due to the heterogeneous nature of bone marrow, experimental results with primary rat bone marrow cells are difficult to reproduce from one experiment to the other, and (2) addition of rhBMP-2 in the medium stimulates the early differentiation and matrix mineralization of osteogenic cells on machined titanium surfaces in a dose-responsive manner. Further, we concluded that our roughened titanium surfaces had no effect on proliferation and differentiation of primary derived rate bone marrow cells.
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PMID:Observations on the effect of BMP-2 on rat bone marrow cells cultured on titanium substrates of different roughness. 1261 75

This study evaluated the shear bond strength of repairs made with a composite (Herculite/Kerr) when two different surface treatment methods and two new generation bond materials were used. The bonding surfaces were prepared by sanding with 500-grit aluminum oxide sandpaper or air abrading with 25-micron aluminum oxide. Treated resin surfaces were acid etched, washed for one minute and air dried. Before adding the composite Herculite, either Optibond Solo (Kerr), Solobond M (Voco) bonding agent or no bonding agent (control group) was applied. The specimens were evaluated for shear bond strength after thermocycling. Fracture surfaces of some samples were also evaluated with SEM. Light microscope and SEM examination of fractured repair surfaces indicated mostly cohesive failure within the air-abraded group. The results showed that surface treatment with air abrasion plus Optibond Solo application had the highest shear bond strength.
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PMID:The effect of air abrasion with two new bonding agents on composite repair. 1267 70

This study evaluated the microtensile bond strength (pTBS) of ethanol/water- and acetone-based, one-bottle adhesive systems to enamel (E) and dentin (D) in the presence (P) or absence (A) of their respective solvents. Thirty-two freshly extracted third molars were flattened with 600-grit SiC paper and restored with Single Bond (SB) or Prime&Bond 2.1 (PB) according to the manufacturers' instructions and after full solvent elimination. The molars were divided into eight test groups (n = 4): G1-SB-E-P, G2-SB-E-A, G3-PBE-P, G4-PB-E-A, G5-SB-D-P, G6-SB-D-A, G7-PB-D-P and G8-PB-D-A. After applying the adhesive resins, composite crowns of approximately 8 mm were built up with TPH Spectrum composite. After 24-hour water storage, the specimens were serially sectioned bucco-lingually to obtain 0.8 mm slabs that were trimmed to an hourglass shape, approximately 0.8 mm2 at the bonded interface. The specimens were tested in tension using a universal testing machine (0.5 mm/minute). The results were statistically analyzed by ANOVA and Tukey test. The frequency of fracture mode was compared using the Kruskal-Wallis test. There were no statistically significant differences in mean bond strength among the groups restored with or without solvent for enamel. However, the results were significantly different for the dentin groups (MPa): G5-26.2 +/- 8.6a; G7-23.6 +/- 11.3ab; G6-12.8 +/- 2.1bc; G8-6.2 +/- 3.1c. SEM examination indicated that the dentin group failure modes were significantly different from the enamel groups. The results suggest that the presence of organic solvents does not influence microTBS to enamel. However, microTBS to dentin was significantly affected by the absence of solvents in the adhesive system.
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PMID:The effect of organic solvents on one-bottle adhesives' bond strength to enamel and dentin. 1465 83

In the present study, the bone response of titanium implants at early bone healing stages, was evaluated using a tensile test. Test surface of coin-shaped cp. titanium implants were standardized by grit blasting with TiO(2), grain size 180-220 microm. The surface topography of the implant specimens was examined by SEM, and by a confocal laser scanner for evaluation of S(a), S(t) and S(dr). The implants were placed onto the leveled site on the tibia of 12 New Zealand White rabbits, 4 implants in each animal. The rabbits were divided into three groups with different observation times i.e. 2, 4 and 6 weeks. The retention of 12 implants were tested by measuring the pull-out force needed to detach the implant from the bone. There was a significant increase in implant retention from 2 to 4 and to 6 weeks healing time (p<0.05). Four implants from each time point were randomly chosen for histological evaluation. The histological appearance of the implant-bone interface at the different healing times showed noticeable differences in the degree of bone healing and maturation, suggesting that, in rabbits, 6 weeks healing time is a suitable observation point for tensile testing of surface optimized osseointegrating implants.
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PMID:Tensile force testing of optimized coin-shaped titanium implant attachment kinetics in the rabbit tibiae. 1534 20

The biocompatibility of material plays an important role in the bone-implant interface for the prosthetic implant fixation. The biocompatibility of implants is associated with the chemical composition, surface topography, surface energy and surface roughness of biomaterials. The effects of two factors, surface roughness and serum contents, on osteoblast behavior at the surface of Ti-6Al-4V and plasma sprayed HA coating were investigated in the experiment. The osteoblasts derived from neonatal rat calvarial were cultured in Dulbecco's modified Eagle medium (DMEM) with fetal bovine serum (FBS) on the surface of polished Ti-6Al-4V (Ti-p), grit-blasted Ti-6Al-4V (Ti-b), polished HA coating (HAC-p), and as-sprayed HA coating (HAC). Under culture medium containing 4% FBS, the level of cell attachment to the polished surface is significantly higher than the rough surface of the same experimental materials during all culture periods. Increasing the contents of FBS up to 10%, the difference of osteoblast attachment is not found between Ti-p and Ti-b. Under 4% serum condition, the cell morphology attached to smooth surfaces (Ti-p and HAC-p) is spread faster and are more flattened than the one to rough surface of the same experimental materials by SEM. After 24 h culture, the corroded cracks are easily observed at the surface of polished HA coatings, and the cell morphology on HAC-p coatings are elongated and less flattened compared with Ti-p. The result is consistent with statistical difference of cell attachment between Ti-p and HAC-p under 4% serum condition.
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PMID:The cell attachment and morphology of neonatal rat calvarial osteoblasts on the surface of Ti-6Al-4V and plasma-sprayed HA coating: effect of surface roughness and serum contents. 1534 8

This study examined the effect of different acidic treatments and the role of a phosphate monomer in a silane coupling agent on the durability of the dual-cure resin cement/silicon oxide bond. Ceramic blocks (Vita Celay Blanks) were cut into multiple 3 mm-thick slices and polished using 600 grit SiC paper. Two pairs were left untreated [controls (CTRL)], two pairs were treated with 40% phosphoric acid and rinsed with water for 30 s (PA), and another two pairs treated with 20% hydrofluoric acid followed by 30 s water rinsing (HF). Half the specimens were silanated with Tokuso Ceramics Primer (TCP) (Tokuyama) and the other half with TCP formulated without phosphate monomer (TCP-NoPM). All the pairs were bonded with Bistite II dual-cure resin cement (Tokuyama) and light cured. After 24 h water storage at 37 degrees C, 0.7 mm-thick slabs were serially sectioned. Immediately, after 6 months and after 1 year of water storage, two slabs were randomly selected from each subgroup, and sliced into beams (6 x 0.7 x 0.7 mm) for the microtensile bond strength (muTBS) test. The muTBS data were statistically analyzed using multiple Wilcoxon Signed Rank tests (p < 0.05). Failure modes were determined using a confocal laser-scanning microscope. Ceramic surface morphology after the different acidic treatments was examined using an SEM. After 1 day, in the case of silane treatment with TCP, there were no significant differences in muTBS between the control and acid-treated groups (p > 0.05), whereas with TCP-NoPM, the muTBS of the control was significantly lower than the acid-treated groups (p < 0.05). All the TCP and acid-treated TCP-NoPM groups exhibited significant reductions in muTBS after 6 months (p < 0.05). After 1 year, the muTBS of the acid-treated TCP groups were not significantly different from the control TCP group (p > 0.05). There was also no significant difference between the HF-treated TCP and TCP-NoPM groups (p > 0.05) after 1 year, all exhibiting greater than 10 MPa tensile bond strength. It is suggested that acidic pretreatment of the ceramic surface does not improve the durability of the dual-cure resin cement/silicon oxide ceramic bond when an acidic phosphate monomer is present as an activator in a ceramic primer.
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PMID:Effect of acidic pretreatment combined with a silane coupling agent on bonding durability to silicon oxide ceramic. 1562 74

The aim of this study was to compare the nanoleakage patterns of the resin-dentin interfaces of three dentin bonding systems at both TEM and field emission in lens SEM (FEI-SEM) levels. A standardized smear layer was created with 180-grit silicon carbide paper (SiC) on dentin disks obtained from 18 noncarious human third molars. Specimens were randomly divided into three groups and bonded with a two-step total etching adhesive (Single Bond, SB), a two-step, self-etching adhesive (Clearfil SE BOND, SEB), and a one-step, self-etching adhesive (XENO III, XEIII). Nanoleakage was evaluated by using an ammoniacal silver-nitrate solution. Specimens were processed for TEM and FEI-SEM observation. The TEM of SB revealed silver deposits in adhesive and hybrid layers (HL). High-magnification FEI-SEM micrographs clearly identified these deposits as spherical clusters mainly associated with nonembedded collagen fibrils. TEM and FEI-SEM examination of SEB revealed some clusters of silver deposits within porosities and small channels of the HL. Additional silver deposits were observed between the peritubular dentin walls and the resin tags. XEIII revealed very fine and diffuse silver grains throughout the entire HL. SEM visualization of nanoleakage at a high level of resolution has not been previously described. FEI-SEM technology supported the TEM visualization with three-dimensional morphological data of the relations between the HL constituents and nanoleakage. The results of the present study confirm the hypothesis that both total- and self-etch adhesives are not able to fully infiltrate the dentin substrate.
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PMID:Nanoleakage within the hybrid layer: a correlative FEISEM/TEM investigation. 1567 95

In dentistry, adhesion promotion with 3-methacryloyloxypropyltrimethoxysilane is usually sufficient, but its hydrolytic stability is a continuous concern. The hydrolytic stability of an alternative, 3-isocyanatopropyltriethoxysilane, was compared with that of conventional 3-methacryloyloxypropyltrimethoxysilane. Two silanes, both in 0.1 and 1.0 vol-% in ethanol-water, were evaluated in the attachment of an experimental bis-phenol-A-diglycidyldimethacrylate (Bis-GMA) resin to grit-blasted (with two different systems) titanium. Silane hydrolysis was monitored by FTIR spectrometry. Bis-GMA resin was applied and photo-polymerized on titanium. The specimens were thermocycled (6000 cycles, 5-55 degrees C). Surface analysis was carried out with scanning electron microscopy. Statistical analysis (ANOVA) showed that the highest shear bond was achieved with 0.1% 3-isocyanatopropyltriethoxysilane (12.5 MPa) with silica-coating, and the lowest with 1.0% 3-methacryloyloxypropyltrimethoxysilane (3.4 MPa) with alumina-coating. The silane, its concentration, and the grit-blasting method significantly affected the shear bond strength (p < 0.05). SEM images indicated cohesive failure of bonding, and, in conclusion, 3-isocyanatopropyltriethoxysilane is a potential coupling agent.
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PMID:Isocyanato- and methacryloxysilanes promote Bis-GMA adhesion to titanium. 1579 Jul 44

This in vitro study morphologically evaluated the effect of some current surface pre-treatments on dentin, using scanning electron microscopy, and related these morphological alterations to clinical implications. The labial surfaces of 30 bovine lower incisors were ground to obtain a flat dentin surface and were finished with 600-grit SiC paper to produce standardized smear layers. The teeth were randomly divided into six groups of five each. Group 1 was the control group, smear layer covered dentin; Group 2 was etched with 37% phosphoric acid (PA) for 15 seconds; Group 3, 37% PA for 15 seconds, followed by 10% NaOCl for 60 seconds; Group 4, 10% NaOCl for 60 seconds; Group 5, a self-etching primer (Clearfil SE Bond, CSEB-primer) was applied for 20 seconds; Group 6, CSEB-primer for 20 seconds, followed by NaOCl for 60 seconds. The specimens were fixed, dehydrated, dried and analyzed by SEM. Treatment with 37% PA removed the smear layer, funneled the tubules and resulted in a collagen-rich surface which appeared to have collapsed in its outermost part, producing a dense surface layer covered with silica particles. When 37% PA treatment was followed by 10% NaOCl, the collagen network was removed to reveal an eroded, rough mineral surface with numerous lateral branches and larger than normal tubular orifices. The action of 10% NaOCl on the smear layer-covered dentin showed no significant alteration in surface morphology. The treatment with CSEB-primer dissolved the smear layer but only partially dissolved the smear plugs. The tubules did not present the typical funnel shape seen following PA treatment. These morphological aspects on dentin surface must influence bonding results. The dentin surface alterations produced by PA appeared to be a very severe demineralization pattern, quite irregular and less permeable to monomer infiltration, while the surface provided by the self-etching primer appeared to be a more uniform, less porous surface, and the association with simultaneous monomer infiltration may reduce the occurrence of mistakes in clinical bonding procedures.
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PMID:The in vitro morphological effects of some current pre-treatments on dentin surface: a SEM evaluation. 1585 6

The purpose of this study was to evaluate the effect of air-powder polishing with sodium bicarbonate (SB) or crystalline cellulose (CC) on the bond strength of a self-etching primer bonding system to dentin. Ground human dentin surfaces were prepared using 600-grit SiC paper. The teeth were divided into three groups according to dentin treatment: control, air-powder polishing with SB, and air-powder polishing with CC. The dentin surfaces were bonded with a self-etching primer bonding system, followed by a light-cured resin composite. microTBS test and SEM analysis were performed. The results of microTBS test indicated that air-powder polishing with SB affected bond strength to dentin, while that with CC did not influence bond strength. SEM observation of air-polished dentin surfaces showed that dentin surfaces air-polished with SB were roughened and covered with a smear layer, while those with CC were smooth and the smear layer was removed.
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PMID:Effect of air-powder polishing on dentin adhesion of a self-etching primer bonding system. 1588 Dec 9


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