Matrix Recruitment and Calcium Sequestration for Spatial Specific Otoconia Development

Otoconia are bio-crystals anchored to the macular sensory epithelium of the utricle and saccule in the inner ear for motion sensing and bodily balance. Otoconia dislocation, degeneration and ectopic calcification can have detrimental effects on balance and vertigo/dizziness, yet the mechanism underlying otoconia formation is not fully understood. In this study, we show that selected matrix components are recruited to form the crystal matrix and sequester Ca2+ for spatial specific formation of otoconia. Specifically, otoconin-90 (Oc90) binds otolin through both domains (TH and C1q) of otolin, but full-length otolin shows the strongest interaction. These proteins have much higher expression levels in the utricle and saccule than other inner ear epithelial tissues in mice. In vivo, the presence of Oc90 in wildtype (wt) mice leads to an enrichment of Ca2+ in the luminal matrices of the utricle and saccule, whereas absence of Oc90 in the null mice leads to drastically reduced matrix-Ca2+. In vitro, either Oc90 or otolin can increase the propensity of extracellular matrix to calcify in cell culture, and co-expression has a synergistic effect on calcification. Molecular modeling and sequence analysis predict structural features that may underlie the interaction and Ca2+-sequestering ability of these proteins. Together, the data provide a mechanism for the otoconial matrix assembly and the role of this matrix in accumulating micro-environmental Ca2+ for efficient CaCO3 crystallization, thus uncover a critical process governing spatial specific otoconia formation

Calcium-deficient apatite: A first in vivo study concerning bone ingrowth

Biphasic calcium phosphate (BCP) materials are increasingly used to restore bone loss in surgery. Calcium-deficient apatites (CDA), the precursors of BCP, are closer in structure to biological apatites and can be associated with therapeutic agents to form drug-delivery systems. The purpose of this first hi vivo study of CDA was to evaluate the osteoconductive properties of two composites, consisting of 40-80 mum granules carried by a cellulose-derived polymer, used to fill critical size bone defects in rabbit femoral ends. Animals were sacrificed 2 or 3 weeks after implantation. Histomorphometric analysis of scanning electron microscopy implant surface files was performed using gray level threshold that distinguish between bone or materials (white) and noncalcified tissue (black). Quantitative results for new bone formation showed no significant differences between the composites or the implantation periods. However, nearly all of the CDA disappeared early while supporting more extensive bone colonization than biphasic calcium phosphates implanted in the same conditions. (C) 2003 Wiley Periodicals, Inc. J Biomed Mater Res 64A: 402-408, 2003

Common SNPs of AmelogeninX (AMELX) and Dental Caries Susceptibility

International audienceGenetic approaches have shown that several genes could modify caries susceptibility; AmelogeninX (AMELX) has been repeatedly designated. Here, we hypothesized that AMELX mutations resulting in discrete changes of enamel microstructure may be found in children with a severe caries phenotype. In parallel, possible AMELX mutations that could explain resistance to caries may be found in caries-free patients. In this study, coding exons of AMELX and exon-intron boundaries were sequenced in 399 individuals with extensive caries (250) or caries-free (149) individuals from nine French hospital groups. No mutation responsible for a direct change of amelogenin function was identified. Seven single-nucleotide polymorphisms (SNPs) were found, 3 presenting a high allele frequency, and 1 being detected for the first time. Three SNPs were located in coding regions, 2 of them being non-synonymous. Both evolutionary and statistical analyses showed that none of these SNPs was associated with caries susceptibility, suggesting that AMELX is not a gene candidate in our studied population

Methods of sample preparation of radula epithelial tissue in chitons (Mollusca: Polyplacophora)

A glutaraldehyde fixative developed for preserving the radula superior epithelium of the adult chiton Acanthopleura hirtosa (Blainville, 1825), was used in conjunction with conventional and microwave-assisted sample processing to produce high quality tissue preservation for light and electron microscopy. In addition, high-pressure freezing (HPF) and cryo-substitution were used to fix the radula tissue of juvenile specimens. Microwave-assisted fixation was preferred to conventional bench-top techniques due to the superior preservation of fine cell structure together with reduced processing times and chemical exposure. Although restricted to very small (<200 μm) samples, the quality of juvenile radulae processed by HPF was excellent. The improvements in tissue preservation using microwave and cryo-preservation techniques are therefore critical for obtaining accurate ultrastructural information on the radula in marine molluscs. In particular, these findings highlight additional processing options available for the study of cellular structures in biomineralizing tissues

S100A8/S100A9 and their association with cartilage and bone

S100A8 and S100A9 are calcium-binding proteins expressed in myeloid cells and are markers of numerous inflammatory diseases in humans. S100A9 has been associated with dystrophic calcification in human atherosclerosis. Here we demonstrate S100A8 and S100A9 expression in murine and human bone and cartilage cells. Only S100A8 was seen in preosteogenic cells whereas osteoblasts had variable, but generally weak expression of both proteins. In keeping with their reported high-mRNA expression, S100A8 and S100A9 were prominent in osteoclasts. S100A8 was expressed in alkaline phosphatase-positive hypertrophic chondrocytes, but not in proliferating chondrocytes within the growth plate where the cartilaginous matrix was calcifying. S100A9 was only evident in the invading vascular osteogenic tissue penetrating the degenerating chondrocytic zone adjacent to the primary spongiosa, where S100A8 was also expressed. Whilst, S100A8 has been shown to be associated with osteoblast differentiation, both S100A8 and S100A9 may contribute to calcification of the cartilage matrix and its replacement with trabecular bone, and to regulation of redox in bone resorption.H. Zreiqat, C. R. Howlett, S. Gronthos, D. Hume and C. L. Gecz