Expression of a Constitutively Active Form of Hck in Chondrocytes Activates Wnt and Hedgehog Signaling Pathways, and Induces Chondrocyte Proliferation in Mice

Runx2 is required for chondrocyte proliferation and maturation. In the search of Runx2 target genes in chondrocytes, we found that Runx2 up-regulated the expression of hematopoietic cell kinase (Hck), which is a member of the Src tyrosine kinase family, in chondrocytes, that Hck expression was high in cartilaginous limb skeletons of wild-type mice but low in those of Runx2?/? mice, and that Runx2 bound the promoter region of Hck. To investigate the functions of Hck in chondrocytes, transgenic mice expressing a constitutively active form of Hck (HckCA) were generated using the Col2a1 promoter/enhancer. The hind limb skeletons were fused, the tibia became a large, round mass, and the growth plate was markedly disorganized. Chondrocyte maturation was delayed until E16.5 but accelerated thereafter. BrdU-labeled, but not terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive, chondrocytes were increased. Furthermore, Hck knock-down reduced the proliferation of primary chondrocytes. In microarray and real-time RT-PCR analyses using hind limb RNA from HckCA transgenic mice, the expression of Wnt (Wnt10b, Tcf7, Lef1, Dkk1) and hedgehog (Ihh, Ptch1, and Gli1) signaling pathway genes was upregulated. These findings indicated that Hck, whose expression is regulated by Runx2, is highly expressed in chondrocytes, and that HckCA activates Wnt and hedgehog signaling pathways, and promotes chondrocyte proliferation without increasing apoptosis

Antxr1, Which is a Target of Runx2, Regulates Chondrocyte Proliferation and Apoptosis

Antxr1/Tem8 is highly expressed in tumor endothelial cells and is a receptor for anthrax toxin.Mutation of Antxr1 causes GAPO syndrome, which is characterized by growth retardation, alopecia,pseudo-anodontia, and optic atrophy. However, the mechanism underlying the growth retardationremains to be clarified. Runx2 is essential for osteoblast di erentiation and chondrocyte maturation and regulates chondrocyte proliferation through Ihh induction. In the search of Runx2 target genes in chondrocytes, we found that Antxr1 expression is upregulated by Runx2. Antxr1 was highly expressed in cartilaginous tissues and was directly regulated by Runx2. In skeletal development, the process of endochondral ossification proceeded similarly in wild-type and Antxr1?/? mice. However,the limbs of Antxr1?/? mice were shorter than those of wild-type mice from embryonic day 16.5 due to the reduced chondrocyte proliferation. Chondrocyte-specific Antxr1 transgenic mice exhibited shortened limbs, although the process of endochondral ossification proceeded as in wild-type mice.BrdU-uptake and apoptosis were both increased in chondrocytes, and the apoptosis-high regions were mineralized. These findings indicated that Antxr1, of which the expression is regulated by Runx2, plays an important role in chondrocyte proliferation and that overexpression of Antxr1 causes chondrocyte apoptosis accompanied by matrix mineralization

Long-term clinical and radiographic evaluation of the effectiveness of direct pulp-capping materials

The aim of this review was to assess the effectiveness of different direct pulp-capping (DPC) materials for human pulp-exposed teeth. An electronic search was performed on 20 February 2018. Long-term clinical and radiographic evaluations of the effectiveness of different DPC materials for use on human pulp-exposed teeth were included. Risk-of-bias assessment and data extraction were performed. From the 496 identified articles, 15 met the eligibility criteria. Among the studies included in those articles, a total of 1,322 teeth were treated with 12 types of DPC materials, and 1,136 teeth were evaluated at a final follow-up examination. For mineral trioxide aggregate (MTA) and calcium hydroxide (CH), the number of included studies, the number of treated teeth, and the mean follow-up period of studies were almost equal, and the success rates of MTA was superior to CH. Therefore, MTA is likely to be a more effective and predictable material for DPC compared to CH. However, the results were based on the included studies, which were all judged to have a high risk of bias. Therefore, more long-term clinical and radiographic studies designed with lower risk of bias are needed. Moreover, the other 10 materials were only investigated by a small number of studies; therefore, further studies are required

Cell migration capability of vascular endothelial growth factor into the root apex of a root canal model in vivo

Once a tooth develops deep caries and the dental pulp tissue is irreversibly infected, the infected dental pulp tissue should be removed, and filling material should be placed in the root canal.Endodontically treated teeth are prone to root fracture or periapical periodontitis; however, dental pulp tissue has the potential to prevent root fracture or periapical periodontitis. Therefore, dental pulp regeneration after pulpectomy may help prolong tooth life. In this study, a new method of dental pulp regeneration was developed. Vascular endothelial growth factor-adsorbed collagen gel was injected into the root canal of a prepared root canal model, placed into the dorsum of a rat, and cultured for 3 weeks. After retrieving the implant, histological analysis was performed.It was found that rat somatic cells were recruited into the root apex of the transplanted root canal model. These findings suggest a new potential technique for engineering dental pulp tissue

Expression of a Constitutively Active Form of Hck in Chondrocytes Activates Wnt and Hedgehog Signaling Pathways, and Induces Chondrocyte Proliferation in Mice

Runx2 is required for chondrocyte proliferation and maturation. In the search of Runx2 target genes in chondrocytes, we found that Runx2 up-regulated the expression of hematopoietic cell kinase (Hck), which is a member of the Src tyrosine kinase family, in chondrocytes, that Hck expression was high in cartilaginous limb skeletons of wild-type mice but low in those of Runx2–/– mice, and that Runx2 bound the promoter region of Hck. To investigate the functions of Hck in chondrocytes, transgenic mice expressing a constitutively active form of Hck (HckCA) were generated using the Col2a1 promoter/enhancer. The hind limb skeletons were fused, the tibia became a large, round mass, and the growth plate was markedly disorganized. Chondrocyte maturation was delayed until E16.5 but accelerated thereafter. BrdU-labeled, but not terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive, chondrocytes were increased. Furthermore, Hck knock-down reduced the proliferation of primary chondrocytes. In microarray and real-time RT-PCR analyses using hind limb RNA from HckCA transgenic mice, the expression of Wnt (Wnt10b, Tcf7, Lef1, Dkk1) and hedgehog (Ihh, Ptch1, and Gli1) signaling pathway genes was upregulated. These findings indicated that Hck, whose expression is regulated by Runx2, is highly expressed in chondrocytes, and that HckCA activates Wnt and hedgehog signaling pathways, and promotes chondrocyte proliferation without increasing apoptosis

Antxr1, Which is a Target of Runx2, Regulates Chondrocyte Proliferation and Apoptosis

Antxr1/Tem8 is highly expressed in tumor endothelial cells and is a receptor for anthrax toxin. Mutation of Antxr1 causes GAPO syndrome, which is characterized by growth retardation, alopecia, pseudo-anodontia, and optic atrophy. However, the mechanism underlying the growth retardation remains to be clarified. Runx2 is essential for osteoblast di erentiation and chondrocyte maturation and regulates chondrocyte proliferation through Ihh induction. In the search of Runx2 target genes in chondrocytes, we found that Antxr1 expression is upregulated by Runx2. Antxr1 was highly expressed in cartilaginous tissues and was directly regulated by Runx2. In skeletal development, the process of endochondral ossification proceeded similarly in wild-type and Antxr1–/– mice. However,the limbs of Antxr1–/– mice were shorter than those of wild-type mice from embryonic day 16.5 due to the reduced chondrocyte proliferation. Chondrocyte-specific Antxr1 transgenic mice exhibited shortened limbs, although the process of endochondral ossification proceeded as in wild-type mice.BrdU-uptake and apoptosis were both increased in chondrocytes, and the apoptosis-high regions were mineralized. These findings indicated that Antxr1, of which the expression is regulated by Runx2, plays an important role in chondrocyte proliferation and that overexpression of Antxr1 causes chondrocyte apoptosis accompanied by matrix mineralization