The effect of aging on the nanostructure of murine alveolar bone and dentin.

INTRODUCTION: Alveolar bone, dentin, and cementum provide a striking example of structurally different collagen-based mineralized tissues separated only by periodontal ligament. While alveolar bone is strongly remodeled, this does not hold for dentin and cementum. However, additional dentin can be deposited on the inner surface of the pulp chamber also in older age. By investigating alveolar bone and molar of mice, the aim of our study is to detect changes in the mineral nanostructure with aging. MATERIALS AND METHODS: Buccal-lingual sections of the mandible and first molar from C57BL/6 mice of three different age groups (young 5 weeks, adult 22 weeks and old 23 months) were characterized using synchrotron small and wide-angle X-ray scattering. Local average thickness and length of the apatite particles were mapped with several line scans covering the alveolar bone and the tooth. RESULTS: In alveolar bone, a spatial gradient was seen to develop with age with the thickest and longest particles in the distal part of the bone. The mineral particles in dentin were found to be become thicker, but then decrease of average length from adult to old animals. The mineral particle characteristics of dentin close to the pulp chamber were not only different to the rest of the tooth, but also when comparing the different age groups and even between individual animals in the same age group. CONCLUSIONS: These results indicated that mineral particle characteristics were found to evolve differently between molar and alveolar bone as a function of age.

Osteoblastic lysosome plays a central role in mineralization.

Mineralization is the most fundamental process in vertebrates. It is predominantly mediated by osteoblasts, which secrete mineral precursors, most likely through matrix vesicles (MVs). These vesicular structures are calcium and phosphate rich and contain organic material such as acidic proteins. However, it remains largely unknown how intracellular MVs are transported and secreted. Here, we use scanning electron-assisted dielectric microscopy and super-resolution microscopy for assessing live osteoblasts in mineralizing conditions at a nanolevel resolution. We found that the calcium-containing vesicles were multivesicular bodies containing MVs. They were transported via lysosome and secreted by exocytosis. Thus, we present proof that the lysosome transports amorphous calcium phosphate within mineralizing osteoblasts.

Small Angle X-ray Scattering and Electron Spin Resonance Spectroscopy Study on Fragrance Infused Cationic Vesicles Modeling Scent-Releasing Fabric Softeners.

Industrially relevant systems for household and personal-care products often involve a large number of components. Such multiple component formulations are indispensable and effective for functionalization of the products, but may simultaneously provide more complex structural features compared to those in ideal systems comprising a smaller number of highly pure substances. Using cryogenic transmission electron microscopy (cryo-TEM), small angle X-ray scattering (SAXS), and electron spin resonance (ESR) spectroscopy, we have investigated effects of fragrance-incorporation into cationic vesicles on their bilayer structures and membrane-membrane interactions. Cationic vesicles were prepared from TEQ surfactant, whose major component was di(alkyl fatty ester) quaternary ammonium methosulfate, and fragrance components, l-menthol, linalool, and d-limonene, were infused into the vesicle membranes to model scent-releasing fabric softeners. The cryo-TEM images confirm formation of multilamellar vesicles (MLVs). Generalized indirect Fourier transformation (GIFT) analysis of the SAXS intensities based on the modified Caillé structure factor model reveals that incorporation of a more hydrophobic fragrance component leads to a more pronounced increase of the surface separation (water layer thickness). Furthermore, the fragrance-infused systems show longer-range order of the bilayer correlations and enhanced undulation fluctuation of the membranes than those in the TEQ alone system. The spin-label ESR results indicate different restricted molecular motions in the TEQ bilayers depending on the labeled position and their marked changes upon addition of the fragrance components, suggesting different mixing schemes and solubilization positions of the fragrance molecules in the TEQ bilayers. The present data have demonstrated how the infused fragrance molecules having different hydrophobicity and molecular architectures into the cationic vesicles affect the membrane structures and the intermembrane interactions, which may provide useful information for precisely controlling a fragrance-releasing property.

The nanostructure of murine alveolar bone and its changes due to type 2 diabetes.

Alveolar bone - the bony ridge containing the tooth sockets - stands out by its remodeling activity where bone is being formed and resorbed at a much higher rate than in any other bony tissue. Teeth that are anchored in the jaw through the periodontal ligament exert very large localized loads during mastication that could lead to a unique adaptation of the collagen/mineral structure in the bone. Our aim was to characterize the nanostructure of alveolar bone and to determine the influence of diabetes on structural characteristics of the mineralized matrix. Using small- and wide-angle X-ray scattering (SAXS/WAXS), we studied a spontaneous diabetic mouse model (KK+) and its corresponding healthy controls (KK-) (n=6) to determine the size and mutual alignment of the mineral nanoparticles embedded in the collagen matrix. On cross-sections (buccal-lingual) of the first molar multiple line scans with a spatial resolution of 30µm were performed on each sample, from the lingual to the buccal side of the mandible. Mineral particle thickness and length are decreasing towards the tooth in both buccal and lingual sides of alveolar bone. While mineral particles are well aligned with the long axis of the tooth on the buccal side, they are in a quarter of the measurements oriented along two preferred directions on the lingual side. These nanostructural differences can be interpreted as the result of an asymmetric loading during mastication, leading to a tilting of the tooth in its socket. In diabetic mice particle thicknesses are smaller compared to control animals.

In vitro analysis of the effect of alkyl-chain length of anionic surfactants on the skin by using a reconstructed human epidermal model.

We investigated the effect of the alkyl-chain length of anionic surfactants on the skin using an in vitro model. The evaluated anionic surfactants were sodium alkyl sulfate (AS) and sodium fatty acid methyl ester sulfonate (MES), which had different alkyl-chain lengths (C8-C14). Skin tissue damage and permeability were examined using a reconstructed human epidermal model, LabCyte EPI-MODEL24. Skin tissue damage was examined by measuring cytotoxicity with an MTT assay. Liquid chromatography/tandem mass spectrometry (LC/MS-MS) and liquid chromatography/mass spectrometry (LC/MS) were used to detect surfactants that permeated into the assay medium through an epidermal model. To assess the permeation mechanism and cell damage caused by the surfactants through the epidermis, we evaluated the structural changes of Bovine Serum Albumin (BSA), used as a simple model protein, and the fluidity of 1,2-dipalmitoyl-sn-glycero-3-phosphpcholine (DPPC) liposome, which serves as one of the most abundant phospholipid models of living cell membranes in the epidermis. The effects of the surfactants on the proteins were measured using Circular Dichroism (CD) spectroscopy, while the effects on membrane fluidity were investigated by electron spin resonance (ESR) spectroscopy. ET50 (the 50% median effective time) increased as follows: C10 < C12 < C8 < C14 in AS and C8, C10 < C12 < C14 in MES. The order of permeation through the LabCyte EPI-MODEL24 was C10 > C12 > C14, for both AS and MES. For both AS and MES, the order parameter, which is the criteria for the microscopic viscosity of lipid bilayers, increased as follows: C10 < C12 < C14, which means the membrane fluidity is C10 > C12 > C14. It was determined that the difference in skin tissue damage in the LabCyte EPI-MODEL24 with C10 to C14 AS and MES was caused by the difference in permeation and cell membrane fluidity through the lipid bilayer path in the epidermis.

Amphiphilic sulfamide as a low-molecular-mass hydrogelator: A novel mode of 3-D networks formed by hydrogen-bond-directed 2-D sheet assemblies.

Asymmetrically substituted amphiphilic sulfamide, N-tetradecyl-N'-(6-dimethylaminohexyl)-sulfamide (3c) having a dimethylamino group at one end of the side chain, showed a strong ability to form two-dimensional (2-D) sheet-like assemblies by the 2-D hydrogen-bond networks between sulfamide moieties. Upon protonation of the amino group with acid, the cationic ammonium form of 3c induced effective hydrogelation (minimum gelation concentration: 0.5wt%) to yield a translucent, self-standing hydrogel. Infrared (IR) spectroscopy, X-ray diffraction (XRD), atomic force microscopy (AFM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) studies confirmed formation of the fibrous assemblies of the hydrogen-bond-directed 2-D nanosheets in the hydrogel. A novel mode of three-dimensional (3-D) networks was formed by branching and recombination of hydrogen-bond networks and knit-like linkages between the assemblies. The storage and loss moduli of the hydrogel (2wt%) were measured to be in the range of 10(2) and 10(3)Pa, showing relatively high mechanical stability.