Transcriptional regulation of FRZB in chondrocytes by Osterix and Msx2.

INTRODUCTION: Osteoarthritis is a common joint disease that causes destruction of articular cartilage and severe inflammation surrounding knee and hip joints. However, to date, effective therapeutic reagents for osteoarthritis have not been developed because the underlying molecular mechanisms are complex. Recent genetic findings suggest that a Wnt antagonist, frizzled-related protein B (FRZB), is a potential therapeutic target for osteoarthritis. Therefore, this study aimed to examine the transcriptional regulation of FRZB in chondrocytes. MATERIALS AND METHODS: Frzb/FRZB expression was assessed by RT-qPCR analyses in murine articular chondrocytes and SW1353 chondrocyte cell line. Overexpression and knockdown experiments were performed using adenovirus and lentivirus, respectively. Luciferase-reporter and chromatin immunoprecipitation assays were performed for determining transcriptional regulation. Protein-protein interaction was determined by co-immunoprecipitation analysis. RESULTS: Frzb was highly expressed in cartilages, especially within articular chondrocytes. Interleukin-1α markedly reduced Frzb expression in articular chondrocytes in association with cartilage destruction and increases in ADAM metallopeptidase with thrombospondin type 1 motif (Adamts) 4 and Adamts5 expression. Bone morphogenetic protein 2 (BMP2) increased FRZB expression in SW1353 cells through Smad signaling. Osterix and msh homeobox 2 (Msx2), both of which function as downstream transcription factors of BMP2, induced FRZB expression and upregulated its promoter activity. Co-immunoprecipitation results showed a physical interaction between Osterix and Msx2. Knockdown of either Osterix or Msx2 inhibited BMP2-dependent FRZB expression. Chromatin immunoprecipitation indicated a direct association of Osterix and Msx2 with the FRZB gene promoter. CONCLUSION: These results suggest that BMP2 regulates FRZB expression through Osterix and Msx2.

Regulatory Mechanisms of Prg4 and Gdf5 Expression in Articular Cartilage and Functions in Osteoarthritis.

Owing to the rapid aging of society, the numbers of patients with joint disease continue to increase. Accordingly, a large number of patients require appropriate treatment for osteoarthritis (OA), the most frequent bone and joint disease. Thought to be caused by the degeneration and destruction of articular cartilage following persistent and excessive mechanical stimulation of the joints, OA can significantly impair patient quality of life with symptoms such as knee pain, lower limb muscle weakness, or difficulty walking. Because articular cartilage has a low self-repair ability and an extremely low proliferative capacity, healing of damaged articular cartilage has not been achieved to date. The current pharmaceutical treatment of OA is limited to the slight alleviation of symptoms (e.g., local injection of hyaluronic acid or non-steroidal anti-inflammatory drugs); hence, the development of effective drugs and regenerative therapies for OA is highly desirable. This review article summarizes findings indicating that proteoglycan 4 (Prg4)/lubricin, which is specifically expressed in the superficial zone of articular cartilage and synovium, functions in a protective manner against OA, and covers the transcriptional regulation of Prg4 in articular chondrocytes. We also focused on growth differentiation factor 5 (Gdf5), which is specifically expressed on the surface layer of articular cartilage, particularly in the developmental stage, describing its regulatory mechanisms and functions in joint formation and OA pathogenesis. Because several genetic studies in humans and mice indicate the involvement of these genes in the maintenance of articular cartilage homeostasis and the presentation of OA, molecular targeting of Prg4 and Gdf5 is expected to provide new insights into the aetiology, pathogenesis, and potential treatment of OA.

Exploring the association between number of teeth, food intake, and cognitive function: A 9-year longitudinal study.

OBJECTIVES: This study aimed to investigate the association between the number of teeth, food intake, and cognitive function in Japanese community-dwelling older adults. METHODS: This 9-year longitudinal study included a total of 293 analyzable participants who participated in baseline and follow-up surveys. Dental status (number of teeth and periodontal pocket depth), dietary assessment using the brief-type self-administered diet history questionnaire, cognitive function, and the following confounding factors were evaluated: educational level, financial satisfaction, living situation, smoking and drinking habits, history of chronic diseases, apolipoprotein E-ε4 carrier, body mass index, handgrip strength, instrumental activities of daily living, and depressive symptomatology. The Japanese version of the Montreal Cognitive Assessment was used to evaluate cognitive function. A multinomial logistic regression analysis for the intake level of each food categorized into three groups (low, moderate, high), and a generalized estimating equation (GEE) for cognitive function over nine years were performed. RESULTS: After controlling for confounding factors, the number of teeth was shown to be associated with the intake of green-yellow vegetables and meat. Furthermore, the GEE indicated that the lowest quartile of intake of green-yellow vegetables significantly associated with lower cognitive function (unstandardized regression coefficient [B] = -0.96, 95 % confidence interval [CI]: -1.72 to -0.20), and the lowest quartile of intake of meat significantly associated with lower cognitive function (B = -1.42, 95 % CI: -2.27 to -0.58). CONCLUSIONS: The intake of green and yellow vegetables and meat, which is influenced by the number of teeth, was associated with cognitive function in Japanese community-dwelling older adults. CLINICAL SIGNIFICANCE: There are few studies that have examined the association between oral health, food intake, and cognitive function. This 9-year longitudinal study suggests that it is important to maintain natural teeth to enable the functional means to consume green-yellow vegetables and meat, and thereby help maintain cognitive function.

Relationship between the number of teeth, occlusal force, occlusal contact area, and dietary hardness in older Japanese adults: The SONIC study.

PURPOSE: Individuals with impaired masticatory function tend to prefer soft foods, which results in decreased masticatory muscle activity. This study examined the association between the oral condition (number of teeth, occlusal force, and occlusal contact area) and dietary hardness using a daily dietary questionnaire. METHODS: This cross-sectional study evaluated 1841 participants aged 69-71 and 79-81 years. Registered dentists examined the number of teeth, occlusal force, and occlusal contact area. Dietary hardness was defined as the estimated masticatory muscle activity required for a habitual diet. Habitual diet during the preceding month was assessed using a brief self-administered diet history questionnaire. Confounding factors, such as age, sex, socioeconomic status, smoking habits, history of chronic diseases (hypertension, hyperlipidemia, and diabetes), and cognitive function were also evaluated. Multivariate linear regression analyses were performed to assess the association between dietary hardness and each oral condition. RESULTS: Occlusal force (standardized regression coefficients [ß]=0.08, P < 0.01) and occlusal contact area (ß=0.06, P < 0.01) were significantly associated with dietary hardness after adjusting for the confounding factors. Number of teeth was not significantly associated with dietary hardness. In addition, the associations between dietary hardness, sex, and a history of diabetes were stronger than those between dietary hardness and oral factors. CONCLUSIONS: Occlusal force and contact area were significantly associated with dietary hardness as estimated from the masticatory muscle activity using a daily diet questionnaire.

Relationship between occlusal force and psychological frailty in Japanese community-dwelling older adults: The Septuagenarians, Octogenarians, Nonagenarians Investigation with Centenarians study.

BACKGROUND: Frailty increases the risk of negative health-related events, such as falls, disability, hospitalizations, and death. Although the association between oral health and physical frailty is well established, the relationship between oral health and psychological frailty has not yet been investigated. Therefore, we conducted a cross-sectional study to examine the association between maximal occlusal force and psychological frailty in Japanese community-dwelling older adults. METHODS: Psychological frailty was defined as a World Health Organization-5 scale (WHO-5) score of <13, cognitive and functional status was defined as a Japanese version of the Montreal Cognitive Assessment (MoCA-J) score of <23, and psychological robustness was defined as a WHO-5 score of ≥13 and a MoCA-J score of ≥23. We used a cross-sectional study design to measure maximal occlusal force in 1810 participants, and examined the following factors relevant to psychological frailty: educational level, financial status, living situation, history of chronic diseases, handgrip strength, and instrumental activities of daily living. We used propensity score matching to match the psychological frailty and psychological robustness groups according to demographic and confounding factors. This process, resulted in 344 participants, of whom 172 were in the psychological frailty group and 172 were in the psychological robustness group. In the matched cohort, differences between groups with and without psychological frailty were compared using generalized estimating equations for maximal occlusal force after adjusting for the number of teeth. RESULTS: After controlling for potential confounding factors of frailty, the psychological frailty group showed lower maximal occlusal force compared with the psychological robustness group (unstandardized regression coefficients = -72.7, 95% confidence interval: -126.3 to -19.1). CONCLUSIONS: Maximal occlusal force was associated with a reduced prevalence of psychological frailty among Japanese community-dwelling older adults participating in our study.

The relationship between changes in occlusal support and masticatory performance using 6-year longitudinal data from the SONIC study.

OBJECTIVES: Reduced occlusal support is thought to be related to a decline in masticatory performance. However, previous research in this field was based on cross-sectional studies. In this study, we conducted a 6-year longitudinal observation of older adults living in the community and examined the associations of changes in occlusal support with masticatory performance. METHODS: Of the 864 participants aged 72-74 years in the SONIC study, 488 who were followed up (median follow-up period 5.92 years) and had no missing data were included in this study. Participants were divided into three groups according to the number of occlusal support zones in the posterior area: Complete occlusion (four zones), Reduced occlusion (one to three zones), and Collapsed occlusion (no occlusal support zone). Longitudinal analysis of the relationship between occlusal support and masticatory performance was undertaken with linear mixed-effects models. RESULTS: Sex, occlusal force, number of unreplaced missing teeth, aging, and occlusal support change were significantly related to masticatory performance. Furthermore, the interaction term between change in occlusal support and aging was a significant explanatory variable for the decline in masticatory performance. The interaction was strongest in the group that changed from Complete or Reduced occlusion to Collapsed occlusion. This result indicates that the loss of occlusal support is a major factor contributing to declining masticatory performance. CONCLUSIONS: The decline of occlusal support was greatly associated with the deterioration of masticatory performance. Our results suggest that older adults need to prevent the collapse of posterior occlusal support to maintain their masticatory performance. CLINICAL SIGNIFICANCE: Occlusal support is important for preserving masticatory performance in older adults. Preventing the loss of molars and retaining occlusal support may contribute to maintaining food intake diversity and nutritional status, thereby improving quality of life. Dental professionals need to carefully examine dental status to assess the risk of occlusal collapse.

Decline of oral functions in old-old adults and their relationship with age and sex: The SONIC study.

BACKGROUND: Many physical functions decline with aging, but it is not known whether oral functions vary according to sex or decline with aging, as it occurs with physical functions. The present study aimed to examine the association of sex, age, and elapsed years with occlusal force and tongue pressure using a generalized linear mixed-effect model (GLMM) over a 3-year period among old-old Japanese adults. METHODS: Participants were community-dwelling older adults who participated in a survey (June 2014-March 2017) and a follow-up survey (July 2017-December 2019) after 3 years (n = 951: 70-year group, n = 466; 80-year group, n = 391; 90-year group, n = 94). Dental examinations including the number of teeth, occlusal force, and tongue pressure were conducted, and a GLMM was used to estimate the association of sex, age, and elapsed years with occlusal force and tongue pressure, adjusting for the number of teeth. RESULTS: The GLMM showed that occlusal force was significantly associated with sex (reference; male, non-standardized coefficient: B = -66.9 [female], p < 0.001), age (reference; 70-year group, B = -81.7 [80-year group], p < 0.001, B = -87.2 [90-year group], p < 0.001), and the number of teeth (B = 13.8, p < 0.001), but did not significantly decrease with elapsed years. Tongue pressure was significantly associated with sex (reference; male, B = -0.94 [female], p = 0.034) and age (reference; 70-year group, B = -1.78 [80-year group], p < 0.001, B = -5.47 [90-year group], p < 0.001). Tongue pressure decreased significantly with elapsed years (B = -0.82, p < 0.001). CONCLUSIONS: These findings suggest that tongue pressure significantly decreased over time, but occlusal force did not. Tongue-related muscles may be more susceptible to aging than masticatory muscles.

Predictive factors for tooth loss in older adults vary according to occlusal support: A 6-year longitudinal survey from the SONIC study.

OBJECTIVES: The aim of this cohort study among community-dwelling older adults aged over 70 years was to investigate the influence of occlusal support on tooth loss, and to determine predictive factors for tooth loss for each occlusal support category using multilevel analyses. METHODS: Participants were 812 older adults who completed the baseline survey and the follow-up survey 6 years later. The Eichner index was used to evaluate occlusal support status. A generalized estimating equation (GEE) logistic regression analysis was used to examine the influence of occlusal support status on tooth loss while adjusting for various factors at individual and tooth levels. Similar analyses were separately performed in each Eichner class to determine predictive factors for tooth loss. RESULTS: The GEE showed that a decline in occlusal support increased the risk of tooth loss (Eichner A: reference category, Eichner B: odds ratio (OR) = 1.96, p < 0.001, Eichner C: OR = 3.04, p < 0.001). Stratified analysis showed that deeper periodontal pockets and abutment teeth for fixed partial dentures were significantly associated with tooth loss, regardless of occlusal support. In Eichner A, the presence of an opposing tooth was advantageous in protecting the tooth, and a missing adjacent tooth was not significantly associated with tooth loss. An opposing tooth was associated with the risk of tooth loss in Eichner B, and a missing adjacent tooth was significantly associated with tooth loss in Eichner B and C. CONCLUSIONS: A decline in occlusal support accelerated tooth loss. Predictive factors for tooth loss varied depending on occlusal support status. CLINICAL SIGNIFICANCE: Occlusal support is an important factor in preventing tooth loss. Dentition conditions such as missing adjacent teeth and the presence of opposing teeth increase the risk of tooth loss in participants with poor occlusal support. Dental personnel need to carefully examine dentition conditions to assess the risk of occlusal collapse.

Smoc1 and Smoc2 regulate bone formation as downstream molecules of Runx2.

Runx2 is an essential transcription factor for bone formation. Although osteocalcin, osteopontin, and bone sialoprotein are well-known Runx2-regulated bone-specific genes, the skeletal phenotypes of knockout (KO) mice for these genes are marginal compared with those of Runx2 KO mice. These inconsistencies suggest that unknown Runx2-regulated genes play important roles in bone formation. To address this, we attempted to identify the Runx2 targets by performing RNA-sequencing and found Smoc1 and Smoc2 upregulation by Runx2. Smoc1 or Smoc2 knockdown inhibited osteoblastogenesis. Smoc1 KO mice displayed no fibula formation, while Smoc2 KO mice had mild craniofacial phenotypes. Surprisingly, Smoc1 and Smoc2 double KO (DKO) mice manifested no skull, shortened tibiae, and no fibulae. Endochondral bone formation was also impaired at the late stage in the DKO mice. Collectively, these results suggest that Smoc1 and Smoc2 function as novel targets for Runx2, and play important roles in intramembranous and endochondral bone formation.

Impact of occlusal force on decline in body mass index among older Japanese adults: Finding from the SONIC study.

OBJECTIVES: To determine any independent influence of occlusal force and of number of natural teeth on decline in body mass index (BMI) among older Japanese adults. DESIGN: Longitudinal study over a 3- to 6-year period. SETTING: Urban and rural area in Japan. PARTICIPANTS: Independently living Japanese adults aged 69-71 years and 79-81 years at baseline. This analysis excluded participants who were defined as underweight at baseline. MEASUREMENTS: Information was collected on age, gender, occlusal force, the number of teeth, BMI, socioeconomic factors, medical history, the number of daily prescription medications, cognitive function, depressive symptoms, hand grip strength, and physical function. Maximal occlusal force was measured with a pressure-sensitive sheet. Nutritional status was assessed using BMI, and participants with BMI <21.5 were defined as underweight. Then, they were divided into two groups: a "BMI declined" group who were defined as underweight at either 3- or 6-year follow-up survey, and a "BMI maintained" group who were not defined as underweight at both follow-up surveys. Logistic generalized estimating equation (GEE) models were used to assess the effect of occlusal force and the number of teeth at baseline on decline in BMI over 3 or 6 years, after adjusting for possible covariates associated with nutritional status. RESULTS: The final analysis included 704 participants. Eighty-six (12.2%) participants were classified into the BMI declined group. Logistic GEE models showed that the number of teeth was not significantly associated with decline in BMI. However, occlusal force was significantly associated with decline in BMI (odds ratio = 0.90, 95% confidence interval = 0.83-0.97) after adjusting for covariates. CONCLUSION: Participants with lower occlusal force were more likely to be in the BMI less than 21.5 kg/m2 . The findings suggest that to prevent decline in oral function is important to maintain nutritional status.

Oral Functions Are Associated with Muscle Strength and Physical Performance in Old-Old Japanese.

Grip strength and walking speed are considered to be important indicators of physical frailty. However, no study has contemporaneously examined any association of multiple oral functions with grip strength and walking speed. The purpose of this study was to examine which oral functions are associated with muscle strength (grip strength), physical performance (walking speed) or both. The study participants were 511 community-dwelling people (254 men and 257 women) aged 77-81 years old. Six oral functions-oral wetness, occlusal force, tongue-lip motor function, tongue pressure, masticatory performance and swallowing function-were measured. Grip strength and walking speed were also measured. A partial correlation analysis, adjusted for gender, showed that occlusal force, tongue-lip motor function, masticatory performance and swallowing function were significantly associated with both grip strength and walking speed. In addition, tongue pressure was significantly associated with grip strength. A general linear model showed that tongue pressure and occlusal force were significantly associated with grip strength. Swallowing function and tongue-lip motor function were significantly associated with walking speed. It is suggested that there are different oral function measures for muscle strength and physical performance, and these oral function measures could be a useful proxy for physical frailty.