PubMed Health⌕ Search

Biomedical subjects

Makoto Hirao

Publications and source records attributed to Makoto Hirao.

5 recordsLinked to original sources
PubMed2006

Oxygen tension regulates chondrocyte differentiation and function during endochondral ossification.

Cartilage functions at a lower oxygen tension than most other tissues. To determine the role of oxygen tension in chondrocyte differentiation and function, we investigated the influence of oxygen tension in the pluripotent mesenchymal cell line C3H10T1/2 and 14.5E mice embryo forelimb organ culture. 10T1/2 cells and embryo forelimbs were cultured under normoxia (20% O2) or hypoxia (5% O2) in the presence of recombinant human bone morphogenetic protein 2. To elucidate the mechanism by which oxygen tension influences chondrocyte differentiation, the Smad pathway was examined using Smad6 overexpression adenovirus and Smad6 transgenic mice embryo forelimbs. The p38 MAPK pathway was examined using dominant-negative MKK3 and FR167653, a specific p38 MAPK inhibitor. The transcriptional activities of Sox9 and Runx2 were also investigated. Hypoxia promoted bone morphogenetic protein 2-induced glycosaminoglycan production and suppressed alkaline phosphatase activity and mineralization of C3H10T1/2. Thus, hypoxia promoted chondrocytic commitment rather than osteoblastic differentiation. In the mice embryo forelimb organ culture, hypoxia increased cartilaginous matrix synthesis. These effects were primarily mediated by p38 MAPK activation, independent of Sox9. Hypoxia inhibited Col10a1 (type X collagen alpha1) expression via down-regulation of Runx2 activity by Smad suppression and histone deacetylase 4 activation. In conclusion, hypoxia promotes chondrocytic differentiation and cartilage matrix synthesis and suppresses terminal chondrocyte differentiation. These hypoxia-induced phenomena may act on chondrocytes to enhance and preserve their phenotype and function during chondrocyte differentiation and endochondral ossification.

Animals↗
PubMed2005

Leptin regulates chondrocyte differentiation and matrix maturation during endochondral ossification.

Leptin has been suggested to mediate a variety of actions, including bone development, via its ubiquitously expressed receptor (Ob-Rb). In this study, we investigated the role of leptin in endochondral ossification at the growth plate. The growth plates of wild-type and ob/ob mice were analyzed. Effects of leptin on chondrocyte gene expression, cell cycle, apoptosis and matrix mineralization were assessed using primary chondrocyte culture and the ATDC5 cell differentiation culture system. Immunohistochemistry and in situ hybridization showed that leptin was localized in prehypertrophic chondrocytes in normal mice and that Ob-Rb was localized in hypertrophic chondrocytes in normal and ob/ob mice. Growth plates of ob/ob mice were more fragile than those of wild-type mice in a mechanical test and were broken easily at the chondro-osseous junction. The growth plates of ob/ob mice showed disturbed columnar structure, decreased type X collagen expression, less organized collagen fibril arrangement, increased apoptosis and premature mineralization. Leptin administration in ob/ob mice led to an increase in femoral and humeral lengths and decrease in the proportional length of the calcified hypertrophic zone to the whole hypertrophic zone. In primary chondrocyte culture, the matrix mineralization in ob/ob chondrocytes was stronger than that of wild-type mice; this mineralization in both types of mice was abolished by the addition of exogenous leptin (10 ng/ml). During ATDC5 cell differentiation culture, exogenous leptin at a concentration of 1-10 ng/ml (equivalent to the normal serum concentration of leptin) altered type X collagen mRNA expression and suppressed apoptosis, cell growth and matrix calcification. In conclusion, we demonstrated that leptin modulates several events associated with terminal differentiation of chondrocytes. Our finding that the growth plates of ob/ob mice were fragile implies a disturbance in the differentiation/maturation process of growth plates due to depletion of leptin signaling in ob/ob mice. These findings suggest that peripheral leptin signaling plays an essential role in endochondral ossification at the growth plate.

Animals↗
PubMed2005

Macro-structural effect of metal surfaces treated using computer-assisted yttrium-aluminum-garnet laser scanning on bone-implant fixation.

Porous coatings have been applied to the surface of prosthetic devices to foster stable device fixation. The coating serves as a source of mechanical interlocking and may stimulate healthy bone growth through osseointegrated load transfer in cementless arthroplasty. Joint arthroplasty by porous-coated prostheses is one of the most common surgical treatments, and has provided painless and successful joint mobility. However, long-term success is often impaired by the loss of fixation between the prosthesis and bone. Porous-coated prostheses are associated with several disadvantages, including metal debris from porous coatings (third body wear particles) and irregular micro-texture of metal surfaces. Consequently, quantitative histological analysis has been very difficult. These issues arise because the porous coating treatment is based on addition of material and is not precisely controllable. We recently developed a precisely controllable porous texture technique based on material removal by yttrium-aluminum-garnet laser. Free shapes can be applied to complex, three-dimensional hard metal surfaces using this technique. In this study, tartan check shapes made by crossing grooves and dot shapes made by forming holes were produced on titanium (Ti6A14V) or cobalt chrome (CoCr) and evaluated with computer-assisted histological analysis and measurement of bone-metal interface shear strength. Width of grooves or holes ranged from 100 to 800 mum (100, 200, 500, and 800 microm), with a depth of 500 microm. When the cylindrical porous-texture-treated metal samples (diameter, 5 mm; height, 15 mm) were implanted into a rabbit femoral condyle, bone tissue with bone trabeculae formed in the grooves and holes after 2 or 4 weeks, especially in 500-microm-wide grooves. Abundant osteoconduction was consistently observed throughout 500-microm-wide grooves in both Ti6A14V and CoCr. Speed of osteoconduction was faster in Ti6A14V than in CoCr, especially in the tartan check shape made of 500-microm-wide grooves. In pushout testing, the tartan check shape made of 500-microm-wide grooves had significantly higher bone-metal interface shear strength than the dot shape or commercial porous coating. These results indicate that the tartan check shape made of 500-microm-wide grooves on metal surfaces has potential for clinical application in artificial prosthesis design.

Aluminum↗
PubMed2005

A new biotechnology for articular cartilage repair: subchondral implantation of a composite of interconnected porous hydroxyapatite, synthetic polymer (PLA-PEG), and bone morphogenetic protein-2 (rhBMP-2).

OBJECTIVE: Articular cartilage repair remains a major obstacle in tissue engineering. We recently developed a novel tool for articular cartilage repair, consisting of a triple composite of an interconnected porous hydroxyapatite (IP-CHA), recombinant human bone morphogenetic protein-2 (rhBMP-2), and a synthetic biodegradable polymer [poly-d,l-lactic acid/polyethylene glycol (PLA-PEG)] as a carrier for rhBMP-2. In the present study, we evaluated the capacity of the triple composite to induce the regeneration of articular cartilage. METHODS: Full-thickness cartilage defects were created in the trochlear groove of 52 New Zealand White rabbits. Sixteen defects were filled with the bone morphogenetic protein (BMP)/PLA-PEG/IP-CHA composite (group I), 12 with PLA-PEG/IP-CHA (group II), 12 with IP-CHA alone (group III), and 12 were left empty (group IV). The animals were killed 1, 3, and 6 weeks after surgery, and the gross appearance of the defect sites was assessed. The harvested tissues were examined radiographically and histologically. RESULTS: One week after implantation with the BMP/PLA-PEG/IP-CHA composite (group I), vigorous repair had occurred in the subchondral defect. It contained an agglomeration of mesenchymal cells which had migrated from the surrounding bone marrow either directly, or indirectly via the interconnecting pores of the IP-CHA scaffold. At 6 weeks, these defects were completely repaired. The regenerated cartilage manifested a hyaline-like appearance, with a mature matrix and a columnar organization of chondrocytes. CONCLUSIONS: The triple composite of rhBMP-2, PLA-PEG, and IP-CHA promotes the repair of full-thickness articular cartilage defects within as short a period as 3 weeks in the rabbit model. Hence, this novel cell-free implant biotechnology could mark a new development in the field of articular cartilage repair.

Animals↗