A FoxA2+ long-term stem cell population is necessary for growth plate cartilage regeneration after injury.

Longitudinal bone growth, achieved through endochondral ossification, is accomplished by a cartilaginous structure, the physis or growth plate, comprised of morphologically distinct zones related to chondrocyte function: resting, proliferating and hypertrophic zones. The resting zone is a stem cell-rich region that gives rise to the growth plate, and exhibits regenerative capabilities in response to injury. We discovered a FoxA2+group of long-term skeletal stem cells, situated at the top of resting zone, adjacent the secondary ossification center, distinct from the previously characterized PTHrP+ stem cells. Compared to PTHrP+ cells, FoxA2+ cells exhibit higher clonogenicity and longevity. FoxA2+ cells exhibit dual osteo-chondro-progenitor activity during early postnatal development (P0-P28) and chondrogenic potential beyond P28. When the growth plate is injured, FoxA2+ cells expand in response to trauma, and produce physeal cartilage for growth plate tissue regeneration.

Overexpression of transcription factor FoxA2 in the developing skeleton causes an enlargement of the cartilage hypertrophic zone, but it does not trigger ectopic differentiation in immature chondrocytes.

We previously found that FoxA factors are necessary for chondrocyte differentiation. To investigate whether FoxA factors alone are sufficient to drive chondrocyte hypertrophy, we build a FoxA2 transgenic mouse in which FoxA2 cDNA is driven by a reiterated Tetracycline Response Element (TRE) and a minimal CMV promoter. This transgenic line was crossed with a col2CRE;Rosa26rtTA/+ mouse line to generate col2CRE;Rosa26rtTA/+;TgFoxA2+/- mice for inducible expression of FoxA2 in cartilage using doxycycline treatment. Ectopic expression of FoxA2 in the developing skeleton reveals skeletal defects and shorter skeletal elements in E17.5 mice. The chondro-osseous border was frequently mis-shaped in mutant mice, with small islands of col.10+ hypertrophic cells extending in the metaphyseal bone. Even though overexpression of FoxA2 causes an accumulation of hypertrophic chondrocytes, it did not trigger ectopic hypertrophy in the immature chondrocytes. This suggests that FoxA2 may need transcriptional co-factors (such as Runx2), whose expression is restricted to the hypertrophic zone, and absent in the immature chondrocytes. To investigate a potential FoxA2/Runx2 interaction in immature chondrocytes versus hypertrophic cells, we separated these two subpopulations by FACS to obtain CD24+CD200+ hypertrophic chondrocytes and CD24+CD200- immature chondrocytes and we ectopically expressed FoxA2 alone or in combination with Runx2 via lentiviral gene delivery. In CD24+CD200+ hypertrophic chondrocytes, FoxA2 enhanced the expression of chondrocyte hypertrophic markers collagen 10, MMP13, and alkaline phosphatase. In contrast, in the CD24+CD200- immature chondrocytes, neither FoxA2 nor Runx2 overexpression could induce ectopic expression of hypertrophic markers MMP13, alkaline phosphatase, or PTH/PTHrP receptor. Overall these findings mirror our in vivo data, and suggest that induction of chondrocyte hypertrophy by FoxA2 may require other factors in addition to Runx2 (i.e., Hif2α, MEF2C, or perhaps unknown factors), whose expression/activity is rate-limiting in immature chondrocytes.

A cross-sectional survey of patient's perception and knowledge of dental implants in Japan.

BACKGROUND: This study aimed to collect data regarding patient perception and knowledge of dental implants. It was conducted with the hope that the data would provide dental professionals and policymakers with a better understanding of ways to promote implant therapy. METHODS: An anonymous online survey with 10 questions was distributed through 12 dental offices in Tokyo and provincial cities in Japan to assess patient perception and knowledge of dental implants. Harvard Medical School's IRB approved this study. RESULTS: We collected data from 1172 patients (59% female, 41% male). The most common perceptions of implant therapy were that it was "expensive," "advanced," and "scary". Patients' implant knowledge came primarily from magazines or books, while professional dental societies/associations were the least sought out source of information. Patients believed that the purpose of dental implants was to avoid dentures and improve chewing function. Their primary concerns about dental implants were the cost and longevity. Approximately 12% of patients with dental implants and 61% of patients without implants did not know that bone grafts may be required and that sedation during surgery was an option. For patients who experienced sedation during the procedure, 60% of them want it for future surgeries. Patients also had limited knowledge of bone-graft materials and the effects of CBCT radiation; 75% of the patients expressed concerns over the safety of graft materials and radiation exposure. For patients with a history of dental implant therapy, 80% of them would recommend dental implants to their family and friends. CONCLUSIONS: Overall, patients' experiences with dental implant therapy were positive, but there was a lack of patient education regarding dental implants and their associated procedures. Dental professionals need to take the initiative to improve patient education.

Differential substrate use in EGF- and oncogenic KRAS-stimulated human mammary epithelial cells.

Many metabolic phenotypes in cancer cells are also characteristic of proliferating nontransformed mammalian cells, and attempts to distinguish between phenotypes resulting from oncogenic perturbation from those associated with increased proliferation are limited. Here, we examined the extent to which metabolic changes corresponding to oncogenic KRAS expression differed from those corresponding to epidermal growth factor (EGF)-driven proliferation in human mammary epithelial cells (HMECs). Removal of EGF from culture medium reduced growth rates and glucose/glutamine consumption in control HMECs despite limited changes in respiration and fatty acid synthesis, while the relative contribution of branched-chain amino acids to the TCA cycle and lipogenesis increased in the near-quiescent conditions. Most metabolic phenotypes measured in HMECs expressing mutant KRAS were similar to those observed in EGF-stimulated control HMECs that were growing at comparable rates. However, glucose and glutamine consumption as well as lactate and glutamate production were lower in KRAS-expressing cells cultured in media without added EGF, and these changes correlated with reduced sensitivity to GLUT1 inhibitor and phenformin treatment. Our results demonstrate the strong dependence of metabolic behavior on growth rate and provide a model to distinguish the metabolic influences of oncogenic mutations and nononcogenic growth.

Association between cognitive health and masticatory conditions: a descriptive study of the national database of the universal healthcare system in Japan.

Cognitive health is subject to decline with increasing numbers of lost teeth which impacts mastication. This study is a descriptive data analysis of the association between masticatory and cognitive conditions using a large database. We obtained the dental and medical records from Japan's universal healthcare system (UHCS) from the national database in 2017. The data from 94% of the Japanese population aged 65 and over is included. It is inclusive of diagnostic codes for various types of cognitive impairment, as well as dental treatment records from 2012 to 2017. The cognitive impairment group was compared to those without a diagnosis of cognitive impairment. Crude odds ratio between loss of mastication with natural teeth (exposure) and cognitive impairments (outcome) were compared. Patients who have lost masticatory function are likely to have cognitive impairment with an odds ratio of 1.89 (p<0.0001) for early elderly (aged 65-75) and 1.33 (p<0.0001) for advanced elderly (over 75). Patients who are edentulous and function with complete dentures are likely to have cognitive impairment with an odds ratio of 2.38 (p<0.0001) and 1.38 (p<0.0001), respectively. The data shows a convincing and significant result of an association between cognitive health and oral health, related to masticatory conditions.