Osteoclast differentiation and characteristic trabecular bone formation during growth plate destruction in osteoprotegerin-deficient mice

Osteoprotegerin (OPG) is an osteoblast-derived secreted member of the tumour necrosis factor receptor superfamily that inhibits osteoclastogenesis. Mice that are OPG-deficient have severe bone loss, including growth plate cartilage destruction. Using OPG-deficient mice as a useful animal model, we attempted to clarify differentiation and ultrastructural features of osteoclasts located on destructed growth plate cartilage and trabecular bone matrix. In the humerus and femur of OPG homozygous (-/-) mice, adjacent to the growth plate cartilage, bone trabeculae without a calcified cartilage core were characteristically formed at the metaphyseal side of the medullary cavity, which resulted in an irregular chondrocyte distribution and arrangement in growth plate cartilage. During growth plate cartilage destruction, osteoclasts positive for tartrate-resistant acid phosphatase showed unusual localization on both type-II collagen-positive cartilage and type-I collagen-positive trabecular bone matrix at the ossification centre of the epiphyseal/metaphyseal border. Although multinucleated osteoclasts were distributed within open lacunar canals in the growth plate, those on uncalcified cartilage matrix lacked a ruffled border. Facing the calcified cartilage matrix within lacunar canals, osteoclasts showed irregularly formed ruffled borders. After growth plate destruction, a thin bone layer was deposited on the remaining cartilage surfaces by invading osteoblasts. Osteoclasts formed prominent ruffled border structures on bone matrix, deposited on the remaining growth plate cartilage. These results suggest that, in OPG (-/-) mice, terminal osteoclast differentiation requires the presence of newly produced bone matrix, as the coupled phenomenon of bone formation and resorption, as well as osteoblast-derived cytokines.     

Osteoclast differentiation at growth plate cartilage-trabecular bone junction in newborn rat femur

Using 3-day-old newborn rats, we examined the differentiation processes of osteoclasts associated with the destruction of the femoral growth plate cartilage and primary trabecular bone. In the growth plate cartilage, thin mineralized areas were detected solely in the longitudinal septal cartilage matrix in the hypertrophic zone, but the transverse septal cartilage matrix between adjacent chondrocytic lacunae within a row of chondrocytes remained unmineralized. The longitudinal septal cartilage between adjacent rows of chondrocytes appeared to persist, forming the walls of opened lacunar canals. Consistent with the removal of the transverse septal cartilage matrix, the longitudinal canals of opened chondrocytic lacunae were deeply invaded by capillary vessels, mononuclear cells and multinucleated pre-osteoclasts lacking a ruffled border. CD34-positive endothelial cells of capillary vessels deeply penetrated into the transverse septal cartilage matrix facing the medullary cavity and the opened chondrocytic lacunae. ED1-positive monocytes/macrophages were distributed at the chondro-osseous junction, but they were distant from the erosive front of the transverse septa. Tartrate-resistant acid phosphatase-positive multinucleated pre-osteoclasts lacking a ruffled border and differentiated osteoclasts with a ruffled border were localized mainly at two locations: the chondro-osseous junction and the growth front of primary bone trabeculae. Osteoclasts were located on the type-I collagen-positive bone trabeculae close to the growth plate, but they appeared to be distant from the type-II collagen-positive cartilage matrix. Even within opened chondrocytic lacunae, when osteoclasts were distant from the cartilage and bone matrix, they lacked polarized cytoplasmic organization and a ruffled border. The osteoclasts located in the remaining septal cartilage also exhibited neither a ruffled border nor a clear zone. Osteoclasts with a prominent ruffled border and clear zone were located in bone matrix covering the remaining septal cartilage. These results suggest that osteoclasts require hydroxyapatite crystals and bone matrix constituents for ruffled border formation and are not involved in resorption of the unmineralized transverse and mineralized longitudinal septal cartilage without covering bone matrix at the chondro-osseous junction.     

Cytochemistry of gastric parietal cells with high-pressure freezing followed by freeze-substitution

Gastric parietal cells were examined for changes in their ultrastructure and distribution of the proton pump during feeding and fasting states in rats. The fundic glands from rats fed ad libitum or fasted with free access to water were cryofixed using high-pressure freezing followed by freeze-substitution in acetone containing osmium or acrolein and then embedded in Epon 812 or Lowicryl K4M resin, respectively. Excellent ultrastructural preservation was achieved. During the feeding state, intracellular canaliculi and numerous microvilli were well developed, while tubulovesicles were poorly developed. In contrast, during the fasting state, the microvilli in the narrowed space of the intracellular canaliculi were tightly packed and the tubulovesicles were enlarged. Ultrathin sections were immunostained with antibodies against the alpha- and beta-subunits of the proton pump, H+ x K(+)-ATPase, using the immunogold method. The labelling was strong and clearly localized in comparison with that obtained using the conventional chemical-fixation method. Each subunit was localized on the membrane of the microvilli, intracellular canaliculi and tubulovesicles. The distribution of subunit proteins varied between the two states. During ad libitum feeding, the immunolabelling was localized strongly on the membranes of the microvilli and intracellular canaliculi. In contrast, the labelling was strong on the tubulovesicle membrane in the fasting state. The results obtained with each anti-subunit antibody by H+ x K(+)-ATPase immunostaining revealed differences in distribution and labelling density between the feeding and fasting states.     

Porcine enamel matrix derivative enhances the formation of reparative dentine and dentine bridges during wound healing of amputated rat molars

We examined the biological effects of porcine enamel matrix derivative (EMD; Emdogain) on the formation of reparative dentine and dentine bridges in rat molars after pulp amputation. The pulp chambers of upper molars of Wistar rats were perforated and the amputated pulp surfaces were directly capped with either EMD or its carrier propylene glycol alginate (PGA) as control. The cavities were then restored with glass-ionomer cement. On post-amputation days 4-30, the dissected maxillae were examined by light and electron microscopy. In PGA-capped pulp, reparative dentine had been formed over the dentine walls under the prepared cavity on day 7 post-amputation and its thickness extended until day 30. On day 30, as well as reparative dentine formation, diffuse calcification had occurred beneath the amputated wound surfaces. Dentine bridge formation under the amputated coronal pulp surface was observed in 18.2% of amputated pulp on day 30. In EMD-capped pulp, reparative dentine had already been formed by odontoblast-like cells over the dentine walls, already on day 4 post-amputation, and its thickness extended until day 30. The Ca and P weight % and Ca/P ratio of reparative dentine matrix were similar to those of pre-existing dentine matrix, and these values were not different between PGA and EMD-capped pulp. Dentine bridge formation was observed in 27.3% of EMD-capped pulp on day 30. Our results suggest that EMD enhances the formation of both reparative dentine and dentine bridges during wound healing of amputated rat molar pulp.     

Morphology and Mechanical Properties in Polymer Blends of Photocurable Polymer and Polycarbonate

Relationships between the morphologies and mechanical properties of binary blends of a photocurable polymer (2‐propenoic acid, (octahydro‐4,7‐methano‐1H‐indenediyl) bis(methylene)ester; DCA) and a linear polymer (poly(4,4′‐cyclohexylidene bisphenol carbonate); PCz) have been investigated. The blend films are prepared by in situ photopolymerization of homogeneous mixtures of a DCA‐monomer and PCz. The phase structure has been converted from a semi‐interpenetrating polymer networks (semi‐IPN) structure to a bicontinuous structure by controlling the cure temperature. Bicontinuous phase‐separated structures can be obtained by curing a wide range of compositions of 17–50 wt.‐% PCz at high temperatures. Miscible semi‐IPN structures are attained by means of photopolymerization below the glass transition temperature of the homogenous mixture before performing photoirradiation, such that magnetic relaxation measurements showed the blend to be miscible in the 10 nm order. The tensile strength and modulus reached a maximum in those blends having an intermediate vague phase structure between semi‐IPN and bicontinuous structures that have a strong interfacial interaction, which leads to incomplete phase decomposition in the PCz‐rich matrix phase. The maximum strength and modulus prepared under optimum condition are inferior to those of the individual components. In contrast, the elongation and break energy are greatly improved in those blends with bicontinuous structures having a diffused phase boundary.

DCA‐rich domain size in bicontinuous structure for DCA/PCz system, as a function of cure temperature; (□) 17 wt.‐% PCz, (○) 30 wt.‐% PCz, and (?) 50 wt.‐% PCz.     

Emerging cell array based on reaction–diffusion

This article demonstrates the self-replication and self-organization phenomena based on a reaction–diffusion mechanism by computer simulation. The simulation model consists of a one-dimensional cell array. Each cell contains two kinds of chemical substances, activator u and inhibitor v, that can generate a reaction–diffusion wave, which is a spatial concentration pattern. The cells are supposed to be divided or deleted depending on the concentrations of chemical substances. We tried several kinds of diffusion coefficient in the model, and in some simulations, a self-replication process and a generating cell array with a metabolic process were observed. By applying the division rule and the apoptosis rule, cell arrays duplicate in two oscillating states, i.e., self-replication processes were observed. By applying a division rule and an annihilation rule, a cell array that has a stable length is generated by changing the cell components, i.e., generating a cell array by a metabolic process was observed. Surprisingly, these two phenomena are realized independently of the initial number of cells. This work was presented in part at the 11th International Symposium on Artificial Life and Robotics, Oita, Japan, January 23–25, 2006     

Self-reconfigurable robots

In this article, the concept of a cellular robot that is capable of reconfiguring itself is reviewed. This "self-reconfigurable (SR) robot" exemplifies a new trend in robotics, indeed, we can now build various kinds of SR robots with off-the-shelf technologies of processors, actuators, and sensors. These SR robots, based on modern mechatronics, are still not as adaptable as the liquid metal robot in The Terminator 2 but are just as flexible as any conventional robots     

Real-time exploration of a multi-robot rescue system in disaster areas

A system procedure is proposed for a multi-robot rescue system that performs real-time exploration over disaster areas. Real-time exploration means that every robot exploring the area always has a communication path to human operators standing by at a base station and that the communication path is configured by ad hoc wireless networking. Real-time exploration is essential in multi-robot systems for USAR (urban search and rescue) because operators must communicate with every robot to support the victim detection process and ad hoc networking is suitable to configure a communication path among obstacles. The proposed system procedure consists of the autonomous classification of robots into search and relay types and behavior algorithms for each class of robot. Search robots explore the areas and relay robots act as relay terminals between search robots and the base station. The rule of the classification and the behavior algorithm refer to the forwarding table of each robot constructed for ad hoc networking. The table construction is based on DSDV (destination-sequenced distance vector) routing that informs each robot of its topological position in the network and other essentials. Computer simulations are executed with a specific exploration strategy of search robots. The results show that a multi-robot rescue system can perform real-time exploration with the proposed system procedure and reduce exploration time in comparison with the case where the proposed scheme is not adopted.     

Web Caching Replacement Algorithm Based on Web Usage Data

Web caching is one of the fundamental techniques for reducing bandwidth usage and download time while browsing the World Wide Web. In this research, we provide an improvement in web caching by combining the result of web usage mining with traditional web caching techniques. Web cache replacement policy is used to select which object should be removed from the cache when the cache is full and which new object should be put into the cache. There are several attributes used for selecting the object to be removed, such as the size of the object, the number of times the object was used, and the time when the object was added into the cache. However, the flaw in these previous approaches is that each object is treated separately without considering the relation among those objects. We have developed a system that can record users’ browsing behavior at the resources level. By using information gathered from this system, we can improve web cache replacement policy so that the number of cache hits will increase, resulting in a faster web browsing experience and less data bandwidth, especially at lower cache storage environments such as on smart phones.     

An analytical study of the effect of a support geometry on frequency shift and support loss of piezoelectric ring-shaped resonators for healthcare and environmental applications

Ring-shaped resonators with one support have been designed in this work. The ring-shaped resonator reacts with a mass perturbation to provide eigenstate or frequency shifts which could transfer to electrical signals by piezoelectric effect. The aforementioned ring-shaped resonator is mainly comprised with a multilayer of Pt/Ti/PZT/Pt/Ti/SiO₂ deposited on the silicon-on-insulator wafer and expected to be a contour mode. In order to estimate the sensitivity of the ring-shaped resonator against the mass perturbation, the theoretical analysis was conducted by ANSYS from two aspects including: (a) the effect of support geometry on frequency shift and support loss (from view point of vibration mode shape); and (b) the mass application methodology. It is found that low-amplitude vibration area is smaller at narrow support, and frequency shift of trapezoid support is higher than that of rectangle support.