Inhibition of histone deacetylases class I improves adipogenic differentiation of human periodontal ligament cells.

Periodontal ligament stem cells (PDLSCs) show plasticity towards the adipogenic lineage; however, little has been done on the participation of epigenetic mechanisms. Histone acetylation is a dynamic process, though balanced by histone acetyltransferases (HATs) and histone deacetylases (HDACs) activities. This process can be halted by HDACs inhibitors, such as trichostatin A (TSA) and valproic acid (VPA). This study aimed to determine the role of HDACs class I in adipogenic differentiation of PDL cells. PDLSCs were treated with TSA at concentrations of 100, 200, and 250 nM, or VPA at 1, 4 and 8 mM. Cell viability was assessed using MTT assays. Gene expression of pluripotency markers (NANOG, OCT4, SOX2), HAT genes (p300, GCN5), and HDACs genes (HDAC1-3) was analyzed by RT-qPCR. Adipogenic differentiation was evaluated via oil red O staining, and acetylation of histone H3 lysine 9 (H3K9ac) was examined by Western blot. VPA treatment resulted in a 60% reduction in cell proliferation, compared to a 50% when using TSA. Cell viability was not affected by either inhibitor. Furthermore, both TSA and VPA induced adipogenic differentiation, through an increase in the deposition of lipid droplets and in GCN5 and p300 expression were observed. Western blot analysis showed that TSA increased H3K9ac levels on adipogenic differentiation of PDLSCs. These findings highlight the potential of HDAC inhibitors as a tool for modulating H3K9 acetylation status and thus influencing adipogenic differentiation of PDLCs.

Mechanical Properties of Poly(Alkenoate) Cement Modified with Propolis as an Antiseptic.

BACKGROUND: We assessed the effect of propolis on the antibacterial, mechanical, and adhesive properties of a commercial poly(alkenoate) cement. METHODS: The cement was modified with various concentrations of propolis, and antibacterial assays were performed against S. mutans by both MTT assays and agar diffusion tests. The compressive, flexural, and adhesive properties were also evaluated. RESULTS: the modified cement showed activity against S. mutans in both assays, although reductions in compressive (from 211.21 to 59.3 MPa) and flexural strength (from 11.1 to 6.2 MPa) were noted with the addition of propolis, while adhesive strength (shear bond strength and a novel pull-out method) showed a statistical difference (p < 0.05). CONCLUSION: the antiseptic potential of modified material against S. mutans will allow this material to be used in cases in which low mechanical resistance is required (in addition to its anti-inflammatory properties) when using atraumatic restorative techniques, especially in deep cavities.

Effects of Neutralization on the Physicochemical, Mechanical, and Biological Properties of Ammonium-Hydroxide-Crosslinked Chitosan Scaffolds.

It has been reported that chitosan scaffolds, due to their physicochemical properties, stimulate cell proliferation in different tissues of the human body. This study aimed to determine the physicochemical, mechanical, and biological properties of chitosan scaffolds crosslinked with ammonium hydroxide, with different pH values, to better understand cell behavior depending on the pH of the biomaterial. Scaffolds were either neutralized with sodium hydroxide solution, washed with distilled water until reaching a neutral pH, or kept at alkaline pH. Physicochemical characterization included scanning electron microscopy (SEM), elemental composition (EDX), Fourier-transform infrared (FTIR) spectroscopy, Raman spectroscopy, thermogravimetric analysis (TGA), and mechanical testing. In vitro cytotoxicity was assessed via dental-pulp stem cells' (DPSCs') biocompatibility. The results revealed that the neutralized scaffolds exhibited better cell proliferation and morphology. It was concluded that the chitosan scaffolds' high pH (due to residual ammonium hydroxide) decreases DPSCs' cell viability.

Dental anomalies in cleft lip and palate: A case-control comparison of total and outside the cleft prevalence.

This study's objective was to compare the total and outside the cleft prevalence of dental anomalies (DA) between patients with cleft lip and palate (CLP) and a control group. This retrospective cross-sectional study was done under a case-control design. The case group consisted of 192 non-syndromic patients with complete CLP, while the control group included 411 patients. All subjects had orthopantomography, intra, and extraoral photographs. The prevalence of dental agenesis, supernumerary teeth, impacted teeth, dental transposition, and microdontia were compared using a chi-squared test (P < .05). Next, a second test was made, but only the anomalies outside the cleft were considered for this study. Total prevalence was 89.1% for cases, and 20.9% for controls (P < .01). The prevalence of each DA was significantly higher for the case group. In the analysis of DAs outside the cleft, the total prevalence was still significantly associated (P < .01); however, only dental agenesis was statistically significant (P < .01). Further analysis found that a high rate of upper premolar absence (P < .01) could explain this event. Patients with CLP have a higher prevalence of DAs compared to controls. After considering only the DAs outside the cleft, the total prevalence remains significantly higher. However, this phenomenon is explained mainly by the elevated prevalence of upper premolars' agenesis. This study's results suggest that environmental factors are behind the high prevalence of DAs in subjects with CLP.

Higher Expression of DNA (de)methylation-Related Genes Reduces Adipogenicity in Dental Pulp Stem Cells.

Obesity is a significant health concern that has reached alarming proportions worldwide. The overconsumption of high-energy foods may cause metabolic dysfunction and promote the generation of new adipocytes by contributing to several obesity-related diseases. Such concerns demand a deeper understanding of the origin of adipocytes if we want to develop new therapeutic approaches. Recent findings indicate that adipocyte development is facilitated by tight epigenetic reprogramming, which is required to activate the gene program to change the fate of mesenchymal stem cells (MSCs) into mature adipocytes. Like adipose tissue, different tissues are also potential sources of adipocyte-generating MSCs, so it is interesting to explore whether the epigenetic mechanisms of adipogenic differentiation vary from one depot to another. To investigate how DNA methylation (an epigenetic mark that plays an essential role in controlling transcription and cellular differentiation) contributes to adipogenic potential, dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PLSCs) were analyzed during adipogenic differentiation in vitro. Here, we show that the capacity to differentiate from DPSCs or PLSCs to adipocytes may be associated with the expression pattern of DNA methylation-related genes acquired during the induction of the adipogenic program. Our study provides insights into the details of DNA methylation during the adipogenic determination of dental stem cells, which can be a starting point to identify the factors that affect the differentiation of these cells and provide new strategies to regulate differentiation and adipocyte expansion.

Similar Features, Different Behaviors: A Comparative In VitroStudy of the Adipogenic Potential of Stem Cells from Human Follicle, Dental Pulp, and Periodontal Ligament.

Dental tissue-derived mesenchymal stem cells (DT-MSCs) are a promising resource for tissue regeneration due to their multilineage potential. Despite accumulating data regarding the biology and differentiation potential of DT-MSCs, few studies have investigated their adipogenic capacity. In this study, we have investigated the mesenchymal features of dental pulp stem cells (DPSCs), as well as the in vitro effects of different adipogenic media on these cells, and compared them to those of periodontal ligament stem cells (PLSCs) and dental follicle stem cells (DFSCs). DFSC, PLSCs, and DPSCs exhibit similar morphology and proliferation capacity, but they differ in their self-renewal ability and expression of stemness markers (e.g OCT4 andc-MYC). Interestingly, DFSCs and PLSCs exhibited more lipid accumulation than DPSCs when induced to adipogenic differentiation. In addition, the mRNA levels of adipogenic markers (PPAR, LPL, and ADIPOQ) were significantly higher in DFSCs and PLSCs than in DPSCs, which could be related to the differences in the adipogenic commitment in those cells. These findings reveal that the adipogenic capacity differ among DT-MSCs, features that might be advantageous to increasing our understanding about the developmental origins and regulation of adipogenic commitment.

Growth differences in patients with dental agenesis, how its location impacts facial morphology.

BACKGROUND/PURPOSE: Non-formation of a tooth impacts the morphology of the alveolar bone, which may, in turn, generate an imbalance in facial growth. This retrospective case-control study aimed to determine whether observable differences exist in the facial growth of patients with dental agenesis relative to complete dentition controls. MATERIALS AND METHODS: The sample comprised 75 patients with dental agenesis, and each case was paired with two controls of the same age and gender (n = 150). All patients were measured cephalometrically (31 variables), and both groups were compared with student's t- or Z-test (P < 0.05). Subsequently, ANOVA or Kruskal-Wallis tests (P < 0.05) were used to compare facial growth depending on the missing tooth's sagittal location in the dental arch (anterior or posterior agenesis); as well as its location in the affected bone (maxillary, mandibular, or both). RESULTS: Four measurements with significant differences were found, whereas ten were found in the sagittal location in the dental arch analysis. Regarding the affected bone, there were no affected variables. CONCLUSION: it was found that patients with dental agenesis show differences in the sagittal growth of the upper jaw and in the position of the lower incisor. In the studied population, these changes are strongly influenced by the sagittal location of the missing tooth, while its location in the jaws does not affect facial growth.

Prevalence of Obesity in Elementary School Children and its Association with Dental Caries.

PURPOSE: To determine any associations between obesity and caries activity in the mixed dentition stage among primary school children in a low-income Mexican primary school. MATERIALS AND METHODS: This cross-sectional study was performed in Mexican schoolchildren aged 8-12 years. The body mass index (BMI) was obtained, and children were classified as overweight/obese considering age and sex. The experience of caries in permanent and temporary dentition was established with the sum of decayed/missing/filled teeth (DMFT or deft index for permanent or temporal dentition); a caries index for all teeth was also considered (total decay [TD]). Mann-Whitney U-test was used to contrast the distribution between sexes of the quantitative variables and to contrast the distribution of each variable per category, the Kruskal-Wallis test was used. Spearman's Rho test was used to establish the correlation between the quantitative variables. Multiple linear regression models were performed to find the relationship between the O'Leary index and the BMI. A Multilayer Perceptron was constructed as follows: (a) dependent variables: deft, DMFT, TD and O'Leary index; (b) factor: BMI; (c) covariable: age. RESULTS: A total of 331 children were included in the study. Dental caries prevalence was 32.4% (95% CI 29.7-35.2), while the mean DMFT was 0.64 (± SD 1.00). Through the Spearmen test, a statistically significant negative correlation was found between BMI-for-age with the total experience of carious lesions (r = -0.127, p = 0.021) and with experience of carious lesions in the deciduous dentition (deft) (r = -0.195, p ≤0.001). But when using the linear and logistic regression models to analyse the relationship with the O'Leary index, BMI was not statistically significant. With the Multilayer Perceptron there appears to be less error in the prediction of deft than the other indexes. CONCLUSIONS: This study confirms the high prevalence of obesity in primary school children. It also shows the scarce association between carious lesions and obesity.

In vitro histomorphometric comparison of dental pulp tissue in different teeth.

BACKGROUND: Dental pulp (DP) represents an accessible and valuable source promising of stem cells for clinical application. However, there are some disadvantages associated with the isolation of dental pulp stem cells (DPSCs), which include the size and weight of the pulp tissue needed to yield sufficient cells for culturing in vitro. Therefore, the objective of this study was to compare in vitro histomorphometry of DP from permanent (premolars, third molar), supernumerary and deciduous teeth of patients between 5 and 25 years old with regards to weight, length, width and the cell density in the four regions of the DP in order to obtain quantitative parameters in a tissue that represents a valuable source of stem cells. METHODS: DPs were obtained from 10 central incisors deciduous, 20 permanent teeth (10 premolars, 10 third molars) and 10 supernumeraries (six mesiodents and four inferior premolar shapes). The pulps were carefully removed, and the entire tissue was weighed. The pulp length and the width were measured with a digital Vernier caliper. The cellular density analysis was performed according to the four regions of the DP (coronal, cervical, medial and apical) in histological slides using photography and the ImageJ® program for quantification. RESULTS: The Pearson correlation test revealed that DP weight among different types of teeth is correlated with age in male patients. A significant positive correlation was noted between length and width of the DP with age in both genders. The mean DP weight for supernumerary and third molar teeth was greater than deciduous and premolar teeth. Finally, the histological analysis showed that the coronal and apical portions of DP in supernumerary and premolar teeth have the highest cell density. CONCLUSIONS: The DP of supernumerary teeth has quantitatively the best morphometric parameters and cell density comparable with the quality of DP obtained from deciduous teeth.

Epigenetic Regulation of Adipogenic Differentiation by Histone Lysine Demethylation.

Obesity is a rising public health problem that contributes to the development of several metabolic diseases and cancer. Adipocyte precursors outside of adipose depots that expand due to overweight and obesity may have a negative impact on human health. Determining how progenitor cells acquire a preadipocyte commitment and become mature adipocytes remains a significant challenge. Over the past several years, we have learned that the establishment of cellular identity is widely influenced by changes in histone marks, which in turn modulate chromatin structure. In this regard, histone lysine demethylases (KDMs) are now emerging as key players that shape chromatin through their ability to demethylate almost all major histone methylation sites. Recent research has shown that KDMs orchestrate the chromatin landscape, which mediates the activation of adipocyte-specific genes. In addition, KDMs have functions in addition to their enzymatic activity, which are beginning to be revealed, and their dysregulation seems to be related to the development of metabolic disorders. In this review, we highlight the biological functions of KDMs that contribute to the establishment of a permissive or repressive chromatin environment during the mesenchymal stem cell transition into adipocytes. Understanding how KDMs regulate adipogenesis might prompt the development of new strategies for fighting obesity-related diseases.

Factors Associated with Dental Pain in Mexican Schoolchildren Aged 6 to 12 Years.

OBJECTIVE: To identify dental pain prevalence and associated factors in Mexican schoolchildren. METHODS: This cross-sectional study included 1,404 schoolchildren aged 6 to 12 years from public schools in the city of Pachuca de Soto, Hidalgo, Mexico. Data were collected through a questionnaire that addressed sociodemographic and socioeconomic factors, eating and dental hygiene habits, and behavior variables. The dependent variable was self-reported dental pain in the 12 months prior to the survey. Data were analyzed using nonparametric statistics and a binary logistical regression model. RESULTS: Dental pain prevalence among the studied children was 49.9%. The variables associated in the final model (p < 0.05) were younger mother's age, higher socioeconomic level, absence of an automobile in the home, fried food, fruit intake, lower tooth brushing frequency, never having used mouthwash or not knowing about it, and parents/guardians with regular to high levels of knowledge about oral health and a regular or good/very good perception of their child's oral health. CONCLUSIONS: One in two children in the study had experienced dental pain in the twelve months prior to the survey. The association of socioeconomic variables with dental pain suggested inequalities among the children in terms of oral health.

Impacted Maxillary Canine Prevalence and Its Association with Other Dental Anomalies in a Mexican Population.

Objective. We quantified the prevalence of impacted maxillary canines (IMC) and their association with other dental anomalies (DAs). Materials and Methods. A retrospective study was done with 860 patients 12 to 39 years of age. The prevalence of IMC was calculated and compared by sex. The sample was divided into a control group and an impaction group, and the prevalence was calculated in both for a series of anomalies: agenesis, supernumerary teeth, shape anomalies of the upper laterals (microdontia, peg and barrel shape, and talon cusp), fusion, gemination, other impacted teeth, transposition, and amelogenesis imperfecta. The prevalence values for both groups were compared (Pearson's χ2 test, p ≤ 0.05). Results. IMC were present in 6.04% of the sample with no difference by sex (p = 0.540). Other DAs occurred in 51.92% of the IMC group and in 20.17% of the controls (p < 0.05). Significant associations (p < 0.05) were identified between IMC and four other DAs: microdontia, barrel shape, other impacted teeth, and transposition. The prevalence of all anomalies was lower in the control group. Conclusion. IMC were seen in 6.04% of patients. Patients with this condition also had a higher prevalence of other DAs. These other anomalies should be used as risk indicators for early diagnosis.