Insulin receptor substrate-2 maintains predominance of anabolic function over catabolic function of osteoblasts

Insulin receptor substrates (IRS-1 and IRS-2) are essential for intracellular signaling by insulin and insulin-like growth factor-I (IGF-I), anabolic regulators of bone metabolism. Although mice lacking the IRS-2 gene (IRS-2−/− mice) developed normally, they exhibited osteopenia with decreased bone formation and increased bone resorption. Cultured IRS-2−/− osteoblasts showed reduced differentiation and matrix synthesis compared with wild-type osteoblasts. However, they showed increased receptor activator of nuclear factor κB ligand (RANKL) expression and osteoclastogenesis in the coculture with bone marrow cells, which were restored by reintroduction of IRS-2 using an adenovirus vector. Although IRS-2 was expressed and phosphorylated by insulin and IGF-I in both osteoblasts and osteoclastic cells, cultures in the absence of osteoblasts revealed that intrinsic IRS-2 signaling in osteoclastic cells was not important for their differentiation, function, or survival. It is concluded that IRS-2 deficiency in osteoblasts causes osteopenia through impaired anabolic function and enhanced supporting ability of osteoclastogenesis. We propose that IRS-2 is needed to maintain the predominance of bone formation over bone resorption, whereas IRS-1 maintains bone turnover, as we previously reported; the integration of these two signalings causes a potent bone anabolic action by insulin and IGF-I

Akt1 in Osteoblasts and Osteoclasts Controls Bone Remodeling

Bone mass and turnover are maintained by the coordinated balance between bone formation by osteoblasts and bone resorption by osteoclasts, under regulation of many systemic and local factors. Phosphoinositide-dependent serine-threonine protein kinase Akt is one of the key players in the signaling of potent bone anabolic factors. This study initially showed that the disruption of Akt1, a major Akt in osteoblasts and osteoclasts, in mice led to low-turnover osteopenia through dysfunctions of both cells. Ex vivo cell culture analyses revealed that the osteoblast dysfunction was traced to the increased susceptibility to the mitochondria-dependent apoptosis and the decreased transcriptional activity of runt-related transcription factor 2 (Runx2), a master regulator of osteoblast differentiation. Notably, our findings revealed a novel role of Akt1/forkhead box class O (FoxO) 3a/Bim axis in the apoptosis of osteoblasts: Akt1 phosphorylates the transcription factor FoxO3a to prevent its nuclear localization, leading to impaired transactivation of its target gene Bim which was also shown to be a potent proapoptotic molecule in osteoblasts. The osteoclast dysfunction was attributed to the cell autonomous defects of differentiation and survival in osteoclasts and the decreased expression of receptor activator of nuclear factor-κB ligand (RANKL), a major determinant of osteoclastogenesis, in osteoblasts. Akt1 was established as a crucial regulator of osteoblasts and osteoclasts by promoting their differentiation and survival to maintain bone mass and turnover. The molecular network found in this study will provide a basis for rational therapeutic targets for bone disorders

Large Gender Gap in Oral Hygiene Behavior and Its Impact on Gingival Health in Late Adolescence

Background: Epidemiologic studies provide broad-based evidence that males are at greater risk of severe periodontal diseases than females. Our recent findings further revealed that male gender was an independent risk factor for gingival bleeding in late adolescents in Japan. Gingival health status has been reported to be affected by oral hygiene behavior. However, gender difference in this regard has not yet been clarified. Methods: We conducted a retrospective review of mandatory medical questionnaires administered as part of a legally required freshman medical checkup between April 2017 and 2019 at the University of Tokyo. Results: Among a total of 9376 sets of responses, chosen subjects were 9098 students aged 17–19. For frequency of daily brushing, males brushed less frequently than females (p < 0.001): 1 time or less (male: 22.9%, female: 11.2%), twice (65.0%, 69.2%), 3 times or more (12.1%, 19.6%). For the duration of brushing each time, males brushed for a shorter period of time than females (p = 0.005): 1 min or less (male: 17.2%, female: 14.1%), 2–3 min (46.9%, 49.2%), 4 min or more (35.9%, 36.7%). (1) Male gender, (2) lower frequency of daily brushing and (3) shorter duration of brushing each time, were significantly associated with the presence of gingival bleeding (p < 0.001 for all). Multivariate regression analysis showed that: (1) male gender (odds ratio 1.29, 95% confidence interval 1.15–1.44); (2) frequency of daily brushing: 1 time or less (2.36, 2.02–2.76), twice (1.45, 1.27–1.67); and (3) brushing duration each time: 1 min or less (1.57, 1.39–1.78), 2–3 min (1.26, 1.14–1.39), were independent risk factors for gingival bleeding (p < 0.001 for all). Conclusions: Males showed poorer oral hygiene behavior than females in late adolescents in Japan. Male gender was an independent risk factor for gingival bleeding, as well as poor oral hygiene behavior

Systemic Disorders Closely Associated with Malocclusion in Late Adolescence: A Review and Perspective

Oral diseases such as dental caries and periodontal disease are reported to be associated with various systemic diseases such as heart disease, respiratory disease, diabetes, rheumatism, and metabolic syndrome, thus increasing the importance of prevention and early treatment [...

Application of induced pluripotent stem cells for cartilage regeneration in CLAWN miniature pig osteochondral replacement model

Introduction: Pluripotent stem cells have an advantage that they can proliferate without reduction of the quality, while they have risk of tumorigenesis. It is desirable that pluripotent stem cells can be utilized safely with minimal effort in cartilage regenerative medicine. To accomplish this, we examined the potential usefulness of induced pluripotent stem cells (iPS cells) after minimal treatment via cell isolation and hydrogel embedding for cartilage regeneration using a large animal model. Methods: Porcine iPS-like cells were established from the CLAWN miniature pig. In vitro differentiation was examined for porcine iPS-like cells with minimal treatment. For the osteochondral replacement model, osteochondral defect was made in the quarters of the anteromedial sides of the proximal tibias in pigs. Porcine iPS-like cells and human iPS cells with minimal treatment were seeded on scaffold made of thermo-compression-bonded beta-TCP and poly-L-lactic acid and transplanted to the defect, and cartilage regeneration and tumorigenesis were evaluated. Results: The in vitro analysis indicated that the minimal treatment was sufficient to weaken the pluripotency of the porcine iPS-like cells, while chondrogenic differentiation did not occur in vitro. When porcine iPS-like cells were transplanted into osteochondral replacement model after minimal treatment in vitro, cartilage regeneration was observed without tumor formation. Additionally, fluorescent in situ hybridization (FISH) indicated that the chondrocytes in the regenerative cartilage originated from transplanted porcine iPS-like cells. Transplantation of human iPS cells also showed the regeneration of cartilage in miniature pigs under immunosuppressive treatment. Conclusion: Minimally-treated iPS cells will be a useful cell source for cartilage regenerative medicine. Keywords: iPS cells, Minimal treatment, Osteochondral replacement model, Cartilage regeneratio

Molecular analysis of a mammary analog secretory carcinoma in the upper lip

Introduction: Mammary analog secretory carcinoma (MASC) is a newly described rare malignancy of the salivary glands characterized by an ETS variant 6 (ETV6)–neurotrophic tyrosine kinase receptor type 3 (NTRK3) fusion gene (EN fusion gene). Presentation of case: We present a case of MASC derived from the left upper lip in a 61-year-old woman. ETV6 rearrangement was detected by fluorescence in situ hybridization (FISH). The presence of EN fusion transcripts was verified by reverse-transcriptase polymerase chain reaction (RT-PCR) and sequencing of the PCR products. Accordingly, this tumor was diagnosed as MASC. Moreover, we performed mutation analysis of the 50 known cancer-related genes using next-generation sequencing. No mutation of cancer-related genes was identified here. Subsequently, the methylation status in promoter region of tumor-suppressor genes, RASSF1A and RARB2, was examined. Both genes have been reported to be methylated in malignant salivary gland tumors, but they were found to be unmethylated. Discussion: Recent studies have demonstrated that distinct types of malignant salivary gland carcinomas are driven by specific, highly recurrent genetic alterations. Detection of molecular abnormalities could be powerful diagnostic tools in the field of salivary gland tumors in near future. Conclusion: We experienced a rare malignant salivary gland carcinoma, MASC. We diagnosed this tumor by molecular approach and subsequently tried to identify novel molecular abnormalities. Although no novel molecular alteration except for EN fusion gene was identified, this result might represent the favorable prognosis of patients with MASC