A Descriptive in vitro Electron Microscopic Study of Acidic Fluoride-Treated Enamel: Potential Anti-Erosion Effects.

This study aimed to investigate the surface zones of acidic fluoride-treated enamel. Human teeth were each divided into three or four enamel specimens that were treated for 10 min with solutions of 0.2 and 0.4% HF (pH 3.09 and 2.94), 1.74% SnF2 (pH 2.9), 0.68% TiF4 (pH 1.6) and 0.84% NaF (pH 4.5). Untreated specimens functioned as negative controls. The microstructure and elemental composition of the surface zones were studied by scanning electron microscopy/energy-dispersive X-ray (EDX) analysis, transmission electron microscopy (TEM) and nanospot-EDX following cross-sectional preparation using focused ion beam technology. TEM/EDX analyses of NaF-treated specimens showed a 500-nm-thick closed surface film containing 20-40 at% (atomic percent) F. HF-treated specimens had a distinct surface film 200-600 nm thick (dense, not globular) containing 45-47 at% F. TiF4-treated specimens had a surface film of 200-300 nm in thickness containing 8-11 at% Ti but no detectable fluoride. SnF2-treated specimens had a modified surface enamel layer varying in thickness from 200 to 800 nm with an inhomogeneous distribution of Sn. Local spots were detected with as high as 8 at% Sn (30 wt%, weight percent). The results suggest that the reaction mechanisms of SnF2 and TiF4 solutions with dental enamel differ from those occurring after enamel exposure to acidulated NaF and HF solutions. While the HF and NaF treatments resulted in the formation of CaF2-like material as shown by EDX, no significant surface fluoridation was found for SnF2 and TiF4 solutions within the TEM/EDX detection limits. These results suggest that the erosion-protective mechanisms of these latter compounds probably relate more to the formation of hardly soluble and acid-resistant reaction surface films and less to surface fluoride incorporation.

Morphological Characterization of the Self-Assembly of Virus Movement Proteins into Nanotubes in the Absence of Virus Particles.

One infection mechanism of plant viruses is the generation of nanotubes by viral movement proteins, allowing cell-to-cell virus particle transport. Previously, it was assumed that viral nanotubes extend directly from the host-cell plasma membrane. In virus-infected plants, these nanotubes reach an extraordinary diameter:length ratio (≈100 nm:µm or mm range). Here, viral nanotubes are produced in a transient protoplast system; the coding sequence for alfalfa mosaic virus movement protein is translationally fused to green fluorescent protein. The maximum extension of viral nanotubes into the culture medium is achieved 24-48 h posttransfection, with lengths in the micro- and millimeter ranges. Scanning electron microscopy and transmission electron microscopy show that strong inhomogeneous viral nanotubes are formed compared to particle-filled systems. The nanotubes have similar length, but fluctuating wall thickness and diameter and are susceptible to entanglement and recombination. Indirect methods demonstrate that movement proteins assemble independently at the top of the nanotube. These viral nanotubes grow distinctly from previously known natural particle-filled systems and are a unique biological tubular nanomaterial that has the potential for micro- or nanoapplications as a mechanically stable structural component.

Investigation of cell-substrate interactions by focused ion beam preparation and scanning electron microscopy.

Cell-substrate interactions, which are an important issue in tissue engineering, have been studied using focused ion beam (FIB) milling and scanning electron microscopy (SEM). Sample cross-sections were generated at predefined positions (target preparation) to investigate the interdependency of growing cells and the substrate material. The experiments focus on two cell culturing systems, hepatocytes (HepG2) on nanoporous aluminum oxide (alumina) membranes and mouse fibroblasts (L929) and primary nerve cells on silicon chips comprised of microneedles. Cross-sections of these soft/hard hybrid systems cannot be prepared by conventional techniques like microtomy. Morphological investigations of hepatocytes growing on nanoporous alumina membranes demonstrate that there is in-growth of microvilli from the cell surface into porous membranes having pore diameters larger than 200 nm. Furthermore, for various cell cultures on microneedle arrays contact between the cells and the microneedles can be observed at high resolution. Based on FIB milled cross-sections and SEM micrographs cells which are only in contact with microneedles and cells which are penetrated by microneedles can be clearly distinguished. Target preparation of biological samples by the FIB technique especially offers the possibility of preparing not only soft materials but also hybrid samples (soft/hard materials). Followed by high resolution imaging by SEM, new insights into cell surface interactions can be obtained.