Trps1 is necessary for normal temporomandibular joint development.

Mutation of the human TRPS1 gene leads to trichorhinophalangeal syndrome (TRPS), which is characterized by an abnormal development of various organs including the craniofacial skeleton. Trps1 has recently been shown to be expressed in the jaw joints of zebrafish; however, whether Trps1 is expressed in the mammalian temporomandibular joint (TMJ), or whether it is necessary for TMJ development is unknown. We have analyzed (1) the expression pattern of Trps1 during TMJ development in mice and (2) TMJ development in Trps1 knockout animals. Trps1 is expressed in the maxillo-mandibular junction at embryonic day (E) 11.5. At E15.5, expression is restricted to the developing condylar cartilage and to the surrounding joint disc progenitor cells. In Trps1 knockout mice, the glenoid fossa of the temporal bone forms relatively normally but the condylar process is extremely small and the joint disc and cavities do not develop. The initiation of condyle formation is slightly delayed in the mutants at E14.5; however, at E18.5, the flattened chondrocyte layer is narrowed and most of the condylar chondrocytes exhibit precocious chondrocyte maturation. Expression of Runx2 and its target genes is expanded toward the condylar apex in the mutants. These observations underscore the indispensable role played by Trps1 in normal TMJ development in supporting the differentiation of disc and synoviocyte progenitor cells and in coordinating condylar chondrocyte differentiation.

Bone marrow-derived HipOP cell population is markedly enriched in osteoprogenitors.

We recently succeeded in purifying a novel multipotential progenitor or stem cell population from bone marrow stromal cells (BMSCs). This population exhibited a very high frequency of colony forming units-osteoblast (CFU-O; 100 times higher than in BMSCs) and high expression levels of osteoblast differentiation markers. Furthermore, large masses of mineralized tissue were observed in in vivo transplants with this new population, designated highly purified osteoprogenitors (HipOPs). We now report the detailed presence and localization of HipOPs and recipient cells in transplants, and demonstrate that there is a strong relationship between the mineralized tissue volume formed and the transplanted number of HipOPs.

Activation of the Wnt/ß-catenin pathway and tissue inhibitor of metalloprotease 1 during tertiary dentinogenesis.

Tertiary dentin is deposited inside teeth after various stimuli and serves as a major defensive wall to preserve pulp cells. However, the molecular mechanisms of the activation of quiescent odontoblasts, immature pulp cells and tertiary dentin formation are still unclear. Therefore, we performed a comprehensive gene expression analysis of pulp cells after cavity preparation of 9-week-old rat molars to clarify the critical molecules in tertiary dentinogenesis. As a result, mRNA expression of various molecules was up- or down-regulated. Notably, several members of the matrix metalloprotease family and their endogenous inhibitors were up-regulated after cavity preparation. In situ hybridization showed that tissue inhibitor of metalloprotease 1 (Timp1) was widely and continuously distributed in the pulp beneath the cavity in vivo. We also observed accumulation of ß-catenin in the pulp cells beneath the cavity by fluorescence immunohistochemistry. Furthermore, Timp1 transcription was repressed by a dominant-negative TCF4 in immature undifferentiated mesenchymal cells, but not altered in mature odontoblast-like cells. These results indicate that cavity preparation may activate the Wnt/ß-catenin pathway and the Wnt/ß-catenin pathway and Timp1 may be correlatively involved in pulp repair. Timp1 might play crucial roles in reactivation of immature pulp cells for tertiary dentinogenesis.

Efficacy of polyphasic calcium phosphates as a direct pulp capping material.

OBJECTIVES: Polyphasic calcium phosphates (Poly-CaP), a complex of hydroxyapatite (HAp) and soluble calcium phosphates including alpha-tricalcium phosphate and tetracalcium phosphate, demonstrate promoting effects on hard tissue formation by osteoblasts. We hypothesized that a Poly-CaP block with a soluble calcium phosphates phase on one side and an insoluble HAp phase on the other side is useful for vital pulp therapy as it may promote dentin regeneration and provide the surface effective to achieve sealing. The purpose of this study was to investigate the efficacy of Poly-CaP as a direct pulp capping material by examining the Ca-release profile, the in vivo ability to induce reparative dentinogenesis, and the bonding of HAp surface with adhesive systems. METHODS: Poly-CaP prepared by annealing crude HAp disc was immersed in buffer solution at pH 7.4 or 4.0, and the concentration of Ca released was measured until 15 days. The pulp of 9-week-old Wister rat molar was exposed and capped with Poly-CaP or HAp block, and dentin bridge formation and pulpal inflammation was evaluated histopathologically after 2 or 4 weeks. Etch & rinse or self-etching adhesive was bonded to HAp surface, and the interface was observed using SEM. RESULTS: Poly-CaP exhibited continuous release of Ca with significantly greater amount than HAp at both pH conditions (P<0.05, Student's t-test). Animal tests demonstrated formation of complete dentin bridge at higher rate for Poly-CaP compared with HAp after 4 weeks (P<0.05, Steel-Dwass test). Impregnation of resin into etched HAp surface, with production of intimate contact at the bonding interface, was seen for all adhesives. CONCLUSIONS: Poly-CaP is a potentially useful material for direct pulp capping with the advantages to promote dentin bridge formation and to provide tight sealing by adhesives.