PubMed · 13438916
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D H MOORE, P H GRIMLEY. 1957-03-25. Problems in methacrylate embedding for electron microscopy.. https://doi.org/10.1083/jcb.3.2.255
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We report the encapsulation of MIN6 cells, a pancreatic beta-cell line, using thermally induced gelable materials. This strategy uses aqueous solvent and mild temperatures during encapsulation, thereby minimizing adverse effects on cell function and viability. Using a 2:1 mixture of PNIPAAm-PEG-PNIPAAm tri-block copolymer and PNIPAAm homopolymer that exhibit reversible sol-to-gel transition at approximately 30 degrees C, gels were formed that exhibit mechanical integrity, and are stable in H(2)O, PBS and complete DMEM with negligible mass loss at 37 degrees C for 60 days. MTT assays showed undetectable cytotoxicity of the polymers towards MIN6 cells. A simple microencapsulation process was developed using vertical co-extrusion and a 37 degrees C capsule collection bath containing a paraffin layer above DMEM. Spherical capsules with diameters ranging from 500 to 900 microm were formed. SEM images of freeze-dried capsules with PBS as the core solution showed homogenous gel capsule membranes. Confocal microscopy revealed that the encapsulated cells tended to form small aggregates over 5 days, and staining for live and dead cells showed high viability post-encapsulation. A static glucose challenge with day-5 cultured microencapsulated cells exhibited glucose-dependent insulin secretion comparable to controls of free MIN6 cells grown in monolayers. These results demonstrate the potential use of these thermo-responsive polymers as cell encapsulation membranes.
OBJECTIVES: Assuming that color changes after aging are related to changes in translucency of materials, the purpose of the present study was to determine the correlation between the changes in color and the changes in scattering and absorption properties after accelerated aging with representative dental esthetic restorative materials: glass ionomer, resin-modified glass ionomer, compomer, and resin composite. METHODS: Color was measured according to the CIELAB color scale in the transmittance and reflectance modes and used to calculate changes in color (deltaE*(ab)), color coordinates (deltaL*, delta a*, and delta b*), translucency parameter (deltaTP), scattering coefficient (deltaS), absorption coefficient (deltaK), and light reflectivity (deltaRI) after accelerated aging. Simple correlations between each pair of the changes in optical values were calculated, and multiple regression analysis was used to determine the parameters influencing the changes in color and color coordinates (p = 0.05). RESULTS: In the resin composite and compomer, deltaS, deltaK, and deltaRI values were approximately zero, whereas deltaS was as high as 8.9 in the glass ionomer. For most comparisons, correlation coefficient (r) was between 0.700 and 0.997. DeltaL* was found to have a major influence on color changes, and deltaS, deltaTP, and deltaRI influenced deltaL*. Therefore, changes in scattering and absorption properties, after aging, were closely correlated with changes in color and color coordinates, especially in glass ionomer-based filling materials.
The purpose of this study was to evaluate the effect of the nanofiller in experimental composites on opacity (contrast ratio). Thirteen experimental composites were prepared with three different sizes of fillers: barium glass minifiller (1 microm; 69-76 wt %), silica microfiller (0.04 microm; 0-6 wt %), and silica nanofiller (7 nm; 0-7 wt %). After disk-type specimens were irradiated with a halogen light curing unit at 500 mW/cm(2) for 30 s, the specimens were aged for 6 h at room conditions and were stored in deionized water for 1, 7, 14, 21, 28, 56, and 84 days. The contrast ratios of the specimens were measured as a function of aging period using a spectrophotometer. The distribution morphology of the filler particles in the resin matrix was also examined using energy-filtering transmission electron microscopy. The experimental composites that contained more than 3% nanofiller had significantly lower contrast ratios (p < 0.05). The composites that contained 6 wt % nanofiller had contrast ratios 34-65% lower than the composite that did not contain nanofiller. The values of the contrast ratio from the composites that excluded microfiller were lower than the values from the composites that included microfiller. From the comparison with the 3 different sizes of filler, the contrast ratio of the composite that contained 70 wt % minifiller and 6 wt % microfiller was the highest, the contrast ratio of the composite that contained only 76 wt % minifiller was the median value, and the contrast ratio of the composite that contained 70 wt % minifiller and 6 wt % nanofiller was the lowest. When the microfiller content was decreased from 6 wt % to 0 wt %, the contrast ratio decreased 6-9%. Energy-filtering transmission electron microscopy images indicated that the contrast ratio of experimental composites is related to the distribution morphology of the filler particles in the resin matrix.