Buccal Epithelial Cells Research Articles

Expression of AMELX, AMBN, ENAM, TUFT1, FAM83H and MMP20 Genes in Buccal Epithelial Cells from Patients with Molar Incisor Hypomineralization (MIH)-A Pilot Study.

Molar incisor hypomineralization (MIH) is a developmental defect that affects the enamel tissue of permanent teeth. Clinicians may observe a range of opacities in the affected teeth, varying from white to creamy, yellow, and brown. Of particular interest is an etiology of MIH that has not been rigorously elucidated. Researchers believe that there are many potential etiological factors with strong genetic and/or epigenetic influence. The primary factors contributing to the risk of MIH development include specific medical conditions and circumstances. These encompass prematurity, cesarean delivery, perinatal hypoxia, and various health issues such as measles, urinary tract infections, otitis media, gastrointestinal disorders, bronchitis, kidney diseases, pneumonia, and asthma. Although genetic research in this area has received substantial attention, the investigation of epigenetic factors remains comparatively underexplored. Special attention is given to genes and their protein products involved in amelogenesis. Examples of such genes are AMELX, AMBN, ENAM, TUFT1, FAM83H, and MMP20. The median relative FAM83H gene expression in the control group was 0.038 (0.031-0.061) and 0.045 (0.032-0.087) in the study group in buccal swabs. The median relative TUFT1 gene expression in the control group was 0.328 (0.247-0.456) and 0.704 (0.334-1.183) in the study group in buccal swabs. Furthermore, children with MIH had significantly higher TUFT1 expression levels compared to the control group (p-value = 0.0043). Alterations in the expression of the TUFT1 and FAM83H genes could be contributing factors to MIH pathogenesis. Nonetheless, further investigation is essential to comprehensively elucidate the roles of all analyzed genes in the pathogenesis of MIH.

Impact of age-related changes in buccal epithelial cells on pediatric epigenetic biomarker research

Cheek swabs, heterogeneous samples consisting primarily of buccal epithelial cells, are widely used in pediatric DNA methylation studies and biomarker creation. However, the decrease in buccal proportion with age in adults remains unexamined in childhood. We analyzed cheek swabs from 4626 typically developing children 2-months to 20-years-old. Estimated buccal proportion declined throughout childhood with both increasing chronological and predicted epigenetic age. However, buccal proportion did not associate with age throughout adolescence. Variability in buccal proportion increased with age through the entire developmental range. These trends held inversely true for neutrophil proportions. Correcting for buccal proportion attenuated the weak association with PedBE age acceleration to non-significance during initial estimation. Notably, correcting for buccal proportion attenuated the association of PedBE age acceleration with obsessive-compulsive disorder and strengthened the association with diurnal cortisol slope. Thus, the age-related change in children’s oral cells is a crucial consideration for cell type-sensitive research.

Cetyltrimethylammonium Chloride (CTAC) and Its Formulated Mouthwash Reduce the Infectivity of Streptococcus mutans and Candida albicans in Mono and Dual State.

Early childhood caries (ECC), a severe form of dental caries, is exacerbated by the synergistic interaction between Streptococcus mutans and Candida albicans, leading to greater disease severity than their individual effects. This underscores the need for more targeted and potent therapeutic alternatives. Given the promising anti-infective properties of quaternary ammonium surfactants (QAS), this study explores the microbicidal properties of one such QAS, cetyltrimethylammonium chloride (CTAC), against both individual- and dual-species cultures of S. mutans and C. albicans for effective ECC treatment. Initially, the minimal inhibitory concentrations (MICs) of CTAC were determined to range from 4 to 8µg/mL against S. mutans, C. albicans, and dual-species cultures. Time-kill kinetics, assessed via spot assays and spectrometry, demonstrated that CTAC completely eradicated both individual- and dual-species cultures within 30min of exposure. Furthermore, at sub-MIC concentrations, CTAC effectively reduced biofilm formation and virulence traits in S. mutans (including acidogenicity and aciduricity) and C. albicans (including yeast-to-hyphal transition and filamentation). To explore therapeutic application, a mouthwash containing CTAC was formulated. The results showed that the formulated CTAC mouthwash was as effective at eradicating pathogens as a commercially available mouthwash containing 0.075% cetylpyridinium chloride (CPC). Moreover, the CTAC mouthwash maintained stable physicochemical characteristics and antimicrobial activity over 4weeks. It exhibited rapid killing activity against pathogens, achieving efficacy within just 2min of exposure. Fluorescence microscopy and SEM micrographs confirmed the strong biofilm eradication potential of the CTAC mouthwash. The non-toxic nature of the formulated mouthwash was validated using human buccal epithelial cells, and in vivo studies further demonstrated that CTAC mouthwash significantly reduced bacterial and fungal loads in Galleria mellonella. Overall, the findings of this study highlight the potential application of QAS-CTAC in the treatment of ECC.

The Effect of Medium-Chain Triglycerides Oil on Candida albicans and Streptococcus mutans in Planktonic and Mucosal Models.

Despite MCT oil's potential antimicrobial benefits for gastrointestinal health, its effects on disrupting cariogenic pathogens on oral mucosal surfaces remain underexplored. This study evaluated the impact of MCT oil on the adhesion and invasion of Candida albicans and Streptoccocus mutans using planktonic and mucosal models. First, a planktonic model was used to assess the impact of various concentrations of MCT on the growth of S. mutans and C. albicans. Subsequently, a mucosal model was established by seeding TR-146 human buccal mucosal epithelial cells on a 3 µm porous transwell membrane, forming an epithelial barrier. MCT oil was then applied to the epithelial barriers in different durations (10, 30, and 60 min). Subsequently, C. albicans and S. mutans were introduced in the transwell and their adherence to the epithelial cells and their transmigration through the barriers was assessed using colony-forming unit counts and the barrier integrity was assessed by trans epithelial electrical resistance (TEER). Furthermore, cytotoxicity of MCT oil on mucosal cells was assessed by AlamarBlue assay. We found that higher MCT concentrations (90% and 100%) significantly inhibited C. albicans and S. mutans growth in planktonic conditions. Additionally, MCT oil reduced S. mutans adhesion to epithelial cells, highlighting its potential to interfere with bacterial attachment and colonization to oral mucosa. However, the oil had limited effects on C. albicans adhesion and transmigration. MCT demonstrated no cytotoxic effects on the viability of epithelial cells. The study findings highlight the potential benefits of MCT oil, particularly in oral bacterial inhibition, for oral health applications.

Effect of Age, Hot Beverages and Tobacco Related Products on Buccal Epithelial Cells of Cigarette Smokers and non-Smokers in Ajman, UAE.

This study aimed to find out the effect of age, hot beverages and tobacco related products on buccal mucosa cells between cigarette smokers and non-smokers in Ajman, UAE. A total of 122 samples were collected, with demographic data including age, hot beverage consumption, cigarette smoking and other tobacco practice using pre-designed questionnaires. Buccal cells were collected, stained, and screened for micronuclei (MN) under a microscope and two evaluators independently assessed all the slides. Among the 122 participants, 61.5% were aged ≤35 years, and 38.5% were aged >35 years. All non-smokers had MN values <10, while 87% of smokers had MN values >10 (p<0.001), with a trend of dose-dependent relationship between cigarette consumption and MN frequency. Similar patterns were observed in individuals using other forms of tobacco, with 97.4% exhibiting MN values >10 (p<0.001). Hot beverage consumption ≥7 cups/day was associated with 87% of subjects having MN values >10, highlighting the pattern of alternative forms of tobacco and high consumption of hot beverages association with elevated MN occurrence. Significant associations were found between MN and variables, except for age. The findings underscore the significance of tobacco and hot beverage consumption in MN occurrence, emphasizing the need to address these behaviors to mitigate genotoxicity and associated health risks. Despite age showing no significant correlation with MN frequency within the studied age range, aging combined with cigarette smoking amplifies genetic damage.<br />.

Synthesis of aromatic glycoconjugates as anti-fungal agents against Candida spp. and assessment of their covalent crosslinking capabilities

Covalent drugs are becoming increasingly attractive in drug discovery, as they can enhance potency and selectivity for their molecular targets. Covalent inhibitors have been investigated for several therapeutic applications, including anti-cancer and anti-infection agents. However, there are only a few examples of covalent inhibitors targeting fungal pathogens. We have previously reported aromatic glycoconjugates (AGCs) capable of inhibiting the adhesion of Candida albicans to buccal epithelial cells. In this work, we synthesize novel derivatives of the AGCs to which we have added reactive functional groups, such as acryloyl and vinyl sulfones, and investigated their antifungal efficacy against Candida spp. Although the compounds were ineffective at clinically relevant concentrations, we found that some of the galactose derivatives featuring reactive groups were amongst the most active, so their ability to crosslink nucleophilic side chains was assessed in model reactions.

Anticariogenic activity of marine brown algae Padina boergesenii and its active components towards Streptococcus mutans.

Streptococcus mutans is a well-recognized bacterium that plays a predominant role in the progression of dental caries. Its pathogenicity is linked to several key characteristics, including the ability to produce organic acids (acidogenicity), thrive in low pH environments (aciduricity), synthesize exopolysaccharides (EPS) via glucosyltransferases, and form retentive biofilms. The treatment of dental caries with conventional antibiotics is often ineffective due to the bacterium's capacity to form recalcitrant biofilms. To address these challenges, strategies that specifically target the pathogen's virulence without affecting its viability have emerged as promising alternatives. In this context, we investigated the anticariogenic properties of the methanolic extract of Padina boergesenii (MEPB). MEPB demonstrated substantial, dose-dependent antibiofilm activity, with a maximum inhibition of 93% at 128 μg/mL, without compromising the viability of S. mutans. Anti-virulence assays using sub-MIC (minimum inhibitory concentration) levels of MEPB showed significant reductions in key virulence factors: 75% reduction in sucrose-dependent adherence, 65% reduction in sucrose-independent adherence, along with notable decreases in acid production, acid tolerance, and water-insoluble (85%) and water-soluble (52%) glucan synthesis. Additionally, MEPB significantly reduced cell surface hydrophobicity (55%) and extracellular DNA (eDNA) production (64%). qPCR analysis corroborated these in vitro findings, revealing that MEPB suppresses the expression of genes involved in S. mutans virulence, particularly genes related to EPS synthesis (gtfB, gtfC & gtfD) biofilm formation(gbpB & gbpC) and two-component regulatory system (vicR) were downregulated. Toxicity testing on human buccal epithelial cells confirmed the non-toxic nature of MEPB, suggesting its safety for potential therapeutic use. Furthermore, GC-MS/MS analysis identified palmitic acid, myristic acid, and stearic acid as the major active constituents of the MEPB extract. Subsequent biofilm inhibitory assays confirmed the potent antibiofilm efficacy of these compounds: palmitic acid (85%), myristic acid (72%) and stearic acid (83%). In conclusion, this study identifies P. boergesenii and its active biomolecules as potential anticariogenic agents, offering an alternative approach to combat dental caries by targeting bacterial virulence mechanisms rather than viability.

Interpretation of shifts in the values ​​of some indices of cytome analysis of buccal epithelial cells

Introduction. Accounting for micronuclei (MN) in the epithelium of the buccal mucosa is widely used to identify human exposure to genotoxic factors. Cytome analysis makes the test more sensitive to detect exposure, but it remains difficult to attribute changes in individual indices to toxicity or genotoxicity. We hypothesized that analysis of the frequencies of different forms of nuclear abnormalities in cells of different degrees of maturity could promote our understanding of the biological meaning of shifts in these indicators. Materials and methods. A cytome analysis was carried out taking into account the degree of maturity of epithelial cells in scrapings of the cheek mucosa in 6–7 years children, and, for comparison, in the urothelium of mice and rats in the control and after the administration of a cystitis inducer and the standard mutagen cyclophosphamide (CP). Results. In scrapings of the buccal mucosa from children, the frequency of cells with condensed chromatin in the nucleus, karyorrhexis and karyopyknosis increased significantly in intermediate cells, and the frequency of binuclear cells (BN), cells with nuclear buds (NB) and karyolysis increased only upon reaching a terminally differentiated state. Analysis of suspension preparations of the bladder epithelium in laboratory animals confirmed the predominant accumulation of BN in the superficial layers. In a model of cystitis in rats caused by a single administration of CP at a dose of 30 mg/kg, a decrease in the frequency of BN was observed at the end of the proliferative phase of post-traumatic epithelial regeneration (14 days after CP administration). After feeding mice with CP at a dose of 1.3 mg/kg/day for 14 days, an increase in the frequency of BN was noted among the most mature cells. Limitation of the study is the lack of assessment of the DNA content in the nuclei of epithelial cells, which did not allow evaluating forms of polyploidy other than abortive cytokinesis (endocycling and endomitosis). Conclusion. There was received confirmation of the relevance of interpretation the increase in the frequency of BNs in buccal epithelial cells in a group of exposed people as a manifestation of genotoxic effects; a decrease in the frequency of BNs in some cases may be associated with post-traumatic regeneration of the epithelium.

Molecular Analysis of Aggregatibacter actinomycetemcomitans ApiA, a Multi-Functional Protein.

Aggregatibacter actinomycetemcomitans ApiA is a trimeric autotransporter outer membrane protein (Omp) that participates in multiple functions, enabling A. actinomycetemcomitans to adapt to a variety of environments. The goal of this study is to identify regions in the apiA gene responsible for three of these functions: auto-aggregation, buccal epithelial cell binding, and complement resistance. Initially, apiA was expressed in Escherichia coli. Finally, wild-type A. actinomycetemcomitans and an apiA-deleted version were tested for their expression in the presence and absence of serum and genes related to stress adaptation, such as oxygen regulation, catalase activity, and Omp proteins. Sequential deletions in specific regions in the apiA gene as expressed in E. coli were examined for membrane proteins, which were confirmed by microscopy. The functional activity of epithelial cell binding, auto-aggregation, and complement resistance were then assessed, and regions in the apiA gene responsible for these functions were identified. A region spanning amino acids 186-217, when deleted, abrogated complement resistance and Factor H (FH) binding, while a region spanning amino acids 28-33 was related to epithelial cell binding. A 13-amino-acid peptide responsible for FH binding was shown to promote serum resistance. An apiA deletion in a clinical isolate (IDH781) was created and tested in the presence and/or absence of active and inactive serum and genes deemed responsible for prominent functional activity related to A. actinomycetemcomitans survival using qRT-PCR. These experiments suggested that apiA expression in IDH781 is involved in global regulatory mechanisms that are serum-dependent and show complement resistance. This is the first study to identify specific apiA regions in A. actinomycetemcomitans responsible for FH binding, complement resistance, and other stress-related functions. Moreover, the role of apiA in overall gene regulation was observed.

Considerations for Cell Type Heterogeneity in Pediatric Salivary DNA Methylation Analyses: Comparison of Reference Panels & Stratification by Estimated Cell Type Proportion.

Saliva is a widely used sample in epigenetic research with children due to its non-invasive nature. Since DNA methylation (DNAm) profile is cell type (CT) specific, salivary DNAm associations with exposures may be influenced by CT compositions, which is highly variable in saliva as it contains immune and buccal epithelial cells (BEC). Reference-based CT deconvolution and statistically adjusting estimated CT in DNAm analyses have become an increasingly common practice. However, careful examinations of how different reference panels may affect DNAm results and alternative approaches (e.g., stratification) are lacking. To scrutinize the best analytical strategies on pediatric salivary DNAm, the current study used 529 salivary DNAm samples obtained from children (mean age = 7.26 years, SD = 0.26 years) in the Family Life Project. Our results showed higher estimated CT variability with child than adult reference panels and highlighted the impact of these estimated CT discrepancies on DNAm associations with social variables (socioeconomic status). Stratifying salivary DNAm samples by BEC proportions detected a larger number of significant associations with biological variables (sex) and tissue-specific effect with cotinine level, a tobacco smoke-exposure biomarker. We provide analytical recommendations for future epigenetic research involving pediatric saliva samples.

The environmental impact on aging: Insights from buccal mucosa and molecular biomarkers

Buccal epithelial cells serve as a primary barrier against the inhalation and ingestion of harmful substances, working alongside immune system cells such as natural killer cells to protect the body from health-damaging factors. These epithelial cells can also be used as an alternative tissue source for monitoring the genotoxic effects of external factors such as chemical exposure. This assessment can be performed using molecular biomarkers of aging, which reflect biological age and indicate cellular aging acceleration due to internal and external damage factors, such as environmental hazards. In contrast to chronological age, which merely reflects the passage of time, biological age accounts for individual variation in aging processes. Molecular biomarkers are crucial for distinguishing between normal and pathological processes in the body and for identifying the effects of external factors such as chemical exposures. The identification of specific biomarkers enhances the ability to detect and monitor adverse biological responses and accelerated aging. This review aims to highlight the routes through which environmental hazards enter the body, the application of buccal epithelial cells in assessing genetic modifications, and the introduction of potential molecular biomarkers. However, further research is necessary to elucidate the roles of these biomarkers in determining aging rates and individual variability. Understanding their implications may also help identify new therapeutic targets for preventing premature aging, treating age-related diseases, and developing potential treatments.

In vitro determination of genotoxicity and cytotoxicity induced by stainless steel brackets with and without surface coating in cultures of oral mucosal cells

BackgroundOrthodontics is a speciality of dentistry that uses a plethora of devices made from myriad materials to manage various malocclusions. Prolonged contact of orthodontic appliances with oral tissues can lead to cellular damage, highlighting the need for biocompatible materials to mitigate health risks.ObjectivesTo analyze the genotoxicity and cytotoxicity produced by metal brackets and coated metallic brackets with polymeric and nanoparticle coatings in oral mucosal cells.Materials & methodsThe current study compares the toxicity of 3 different types of orthodontic brackets with control groups of oral mucosal cells. Each of the three treatment groups consisted of 10 samples of orthodontic brackets: stainless steel brackets(Group 1), nanoparticle-coated brackets(Group 2), and polymeric-coated brackets(Group 3) exposed to corrosion eluates employing an oral biomimicry model. Two types of oral mucosal cells- Human Gingival Fibroblasts and Buccal Epithelial Cells were used to study the cytotoxic and/or genotoxic effects of the elutes. Intergroup comparisons were conducted using one-way analysis of variance, while scanning electron microscopy evaluated surface characteristic.ResultsThe interaction between metal ions and oral mucosal cells showed no statistically significant difference for toxicity assays between the three groups(p > 0.005). However, polymeric and nanoparticle-coated groups showed reduced cellular differentiation when compared with conventional stainless-steel brackets.ConclusionThis in-vitro study shows that polymeric or nanoparticle coating of conventional metal brackets aids in enhancing corrosion-resistant characteristics of orthodontic appliances and reduces the toxic oral environment created by metal release in the oral cavity.

Approaches to Identifying Markers of Effect of Environment and Nutrition in Preschoolers

Background: Identification of the markers of effect of environment and nutrition on the human body, given their interplay, remains relevant for health risk assessment. Objective: To identify markers of children’s health effects of environmental exposures and nutrition with account for their mutual influences. Materials and methods: The study involved 197 preschool children living in two towns of the Sverdlovsk Region with different levels of environmental pollution. In January 2022 to June 2024, we tested mass concentrations of 19 metals, cytogenetic parameters of buccal epithelium, interleukins (IL-1, IL-4), glutathione-S-transferase, and 60 organic acids in urine of the subjects. We also evaluated the preschool menu and off-school nutrition, anthropometric parameters, morbidity based on outpatient cards, and health status using a questionnaire-based survey. Microsoft Excel and IBM SPSS were used for statistical data analysis. Results: We established that in the children living in the territory with high airborne risks, the levels of exposure to seven chemicals exceeded the reference values, especially those of aluminum and manganese by 3.9 and 2.5 times, respectively. Cytogenetic damage to buccal epithelial cells, higher values of markers of allergic reactions (IL 4), higher values of glutathione-S-transfer and organic acids, and markers of detoxification were also more frequent in this group (p &lt; 0.001). We noted that the diet of children was deficient in vitamins B1 and C, calcium, and, judging by the markers of organic acids, essential amino acids and vitamins B9 and B12. The exposed children differed from the controls in high exposure to arsenic, mercury, and copper; there was also a larger proportion (85 %) of children with cells with micronuclei and atypical nuclei, high concentrations of inflammatory cytokines (IL 1), energy and protein deficiencies, excess sugars in the diet, and markers of energy metabolism disorders among them. Conclusions: The identified markers of two non-adaptive metabolotypes to environmental exposure and nutritional phenotypes will allow a differentiated approach to developing diets.

Impact of FokI (rs2228570) and BglI (rs739837) polymorphisms in VDR gene on permanent tooth eruption: A cross-sectional study

IntroductionGenetic polymorphisms who disturb the mineral homeostasis during tooth development and eruption are candidate to clarify the molecular mechanisms involved in changes in the tooth eruption chronology. In this study, we evaluate whether the FokI (rs2228570) and BglI (rs739837) polymorphisms in the Vitamin D receptor (VDR) gene are associated with changes in the chronology of eruption of permanent teeth. Material & methodThis cross-sectional study randomly included 353 biologically unrelated children, both sexes, without systemic impairment or syndromes and history of trauma during the primary dentition. One operator perform the oral clinical examination. The tooth was considered erupted if there was a visible minimum of any tooth surface emerging from the mucosa. Genomic DNA was extracted from buccal epithelial cells from saliva samples. Genotyping was performed by Real-Time Polymerase Chain Reactions using TaqMan® technology. The average of the total number of erupted permanent teeth between the genotypes was compared by the Mann-Whitney test and multivariate Generalized Linear Models (GLM) (α = 5 %). β values with Confidence Interval (CI) 95 % were calculated. ResultsThe heterozygous adenine-guanine genotype of the FokI significantly decreases the number of erupted permanent teeth (β = −1.15; CI 95 % = −2.22 to −0.07; p = 0.036). In the stratified analysis for maxillary and mandibular teeth, this genotype was associated with a decrease in the number of erupted maxillary permanent teeth (β = −0.65; CI 95 % = −1.22 to −0.09; p = 0.023). BglI was not associated with permanent teeth eruption. ConclusionThe FokI, but not BglI, in the VDR may delay the eruption of permanent teeth.

Analysis of Mutational Burden of Mitochondrial Genome in Cells of Different Human Organs and Tissues.

Cells of different human organs and tissues contain different numbers of mitochondria. In these organelles, there are different copies of the mitochondrial genome, which is characteristic of a certain organ or tissue. The aim of the investigation was to analyze the results of scientific works dedicated to the analysis of heteroplasmy levels of mitochondrial genome mutations in a number of organs and tissues. Based on literature data, the level of heteroplasmy of mitochondrial genome mutations was analyzed in organs such as the liver, lungs, muscles, small intestine, large intestine, spleen, kidney, brain, heart, and hair. In addition, this parameter was studied in such tissues as leukocytes, buccal epithelium, and epithelial cells from urine. Significant differences in the mutational burden of the mitochondrial genome were found in various samples of organs and tissues. The highest heteroplasmy level for mtDNA mutations was in muscles; it was lower in buccal epithelium; and in human blood cells, the heteroplasmy level of mitochondrial mutations turned out to be significantly lower compared to other tissues. During the comparison of samples of patients with different diseases and healthy people, significant differences were found in the heteroplasmy level between some organs and tissues. The heteroplasmy level of mitochondrial genome mutations can significantly differ in the organs and tissues of individuals. In addition, in a number of literature sources, it is noted that there is a dependence on the mutational burden of the mitochondrial genome from the type of disease, sex, and age of a person.

Sex-Specific Associations between Prenatal Exposure to Bisphenols and Phthalates and Infant Epigenetic Age Acceleration.

We examined whether prenatal exposure to two classes of endocrine-disrupting chemicals (EDCs) was associated with infant epigenetic age acceleration (EAA), a DNA methylation biomarker of aging. Participants included 224 maternal-infant pairs from a Canadian pregnancy cohort study. Two bisphenols and 12 phthalate metabolites were measured in maternal second trimester urines. Buccal epithelial cell cheek swabs were collected from 3 month old infants and DNA methylation was profiled using the Infinium MethylationEPIC BeadChip. The Pediatric-Buccal-Epigenetic tool was used to estimate EAA. Sex-stratified robust regressions examined individual chemical associations with EAA, and Bayesian kernel machine regression (BKMR) examined chemical mixture effects. Adjusted robust models showed that in female infants, prenatal exposure to total bisphenol A (BPA) was positively associated with EAA (B = 0.72, 95% CI: 0.21, 1.24), and multiple phthalate metabolites were inversely associated with EAA (Bs from -0.36 to -0.66, 95% CIs from -1.28 to -0.02). BKMR showed that prenatal BPA was the most important chemical in the mixture and was positively associated with EAA in both sexes. No overall chemical mixture effects or male-specific associations were noted. These findings indicate that prenatal EDC exposures are associated with sex-specific deviations in biological aging, which may have lasting implications for child health and development.

Telehealth Parenting Program and Salivary Epigenetic Biomarkers in Preschool Children With Developmental Delay

Children with developmental delays are at a heightened risk of experiencing mental health challenges, and this risk is exacerbated among racially minoritized children who face disproportionate adversity. Understanding the impact of parenting interventions on biological markers associated with these risks is crucial for mitigating long-term health disparities. To examine the effect of 20 weeks of an internet-based parent-child interaction training (iPCIT) program on biomarkers associated with aging and chronic inflammation among preschoolers with developmental delay at 12-month follow-up. An observational secondary analysis of data from a randomized clinical trial conducted from March 17, 2016, to December 15, 2020, to assess changes in salivary DNA methylation (DNAm)-derived biomarkers following iPCIT intervention. Participants were recruited from 3 Part C early intervention sites in a large southeastern US city. Eligible participants included children recruited within 3 months of their third birthday who had a Child Behavior Checklist Externalizing Problems T score greater than 60 and provided saliva in at least 1 study wave. Data analysis was conducted May 2023 to April 2024. Participants received either iPCIT (a telehealth therapeutic intervention focused on enhancing the parent-child relationship and addressing behavioral challenges in young children) or referrals as usual. DNAm at the 12-month follow-up was assessed using the Infinium HumanMethylationEPIC Bead Chip Assay to derive biomarkers DunedinPACE, C-reactive protein (CRP), and interleukin-6 (IL-6). Analyses were intent-to-treat and used path analysis. A total of 71 children (mean [SD] age, 36.27 [0.61] months 51 male [71.8%] and 20 female [28.2%]) were analyzed, of whom 34 received iPCIT and 37 received referrals as usual. The iPCIT group had a slower pace of aging (β = 0.26; 95% CI, 0.06 to 0.50; P = .03) and less DNAm-derived CRP (β = 0.27; 95% CI, 0.05 to 0.49; P = .01) relative to the control condition at the 12-month follow-up. These associations remained significant after accounting for baseline DNAm score, child demographics, and symptom severity, and were independent of predicted buccal epithelial cell proportion for both DunedinPACE and CRP. There was no association with DNAm-derived IL-6 (β = 0.14; 95% CI, -0.08 to 0.36; P = .21). In this study of a parenting intervention, iPCIT, the association of intervention with decreased molecular markers of inflammation and biological aging suggests their potential to modify aspects of the biological embedding of stress. Understanding the systemic biological impact of such interventions offers insights into addressing health disparities and promoting resilience among vulnerable populations. ClinicalTrials.gov Identifier: NCT03260816.

Micronuclei as an indicator of genotoxic change in epithelial cells of buccal mucosa after panoramic radiographs.

The radiation released at the time of dental panoramic radiographs causes genotoxic and cytotoxic effects on epithelial cells. This research aimed to evaluate the changes in the frequencies of micronucleated cells in patients' buccal epithelial cells following dental panoramic radiography. 74 patients were recruited for the study who were advised for panoramic radiographs. Using a wooden spatula, the buccal epithelial cells were scraped from both cheeks before to panoramic radiation exposure and ten days after the panoramic radiation exposure. Giemsa stain was used to stain the cells, and 500 cells were scored on a slide to determine the frequency of micronuclei. To determine the difference between the frequency of micronuclei before and after radiation exposure, a paired t-test was used in the statistical analysis. The proportion of micronuclei cells was 0.11% before radiation exposure and 0.57% following radiation exposure after 10 days. A statistically significant increase in the frequencies of micronuclei was noted after radiation exposure values. This study revealed the genotoxicity of epithelial cells with dental panoramic radiation exposure. It is advised to reduce the use of such radiographs and to use only when there is no other diagnostic tool that is helpful or when absolutely essential.

Study of the epithelial state of the oral mucosa in patients with recurrent aphthous stomatitis

Relevance. Recurrent aphthous stomatitis (RAS) is one of the most common diseases affecting the oral mucosa. It is characterized by the recurrent appearance of ulcers (aphthae) and a prolonged course with periodic exacerbations.Purpose. To study the condition of the oral mucosa epithelium in patients with recurrent aphthous stomatitis undergoing treatment with antiseptics and reparative agents.This study examined samples of buccal epithelium from 64 patients, who were randomly divided into two equal groups: the first group used chlorhexidine as a mouthwash (control group); the second group used octenidine + phenoxyethanol for 7 days. For pain relief, patients in both groups applied a gel containing lidocaine + chamomile extract. After 7 days, the therapy continued with reparative agents (a hydrogel-based material with sodium alginate, Derinat, and lidocaine, deproteinized calf blood extract, and sea buckthorn oil). Each group was further divided into three subgroups for this part of the therapy. The condition of the buccal epithelium was assessed three times: before treatment, on the 7th day, and on the 21st day of therapy. Buccal epithelium preparations were prepared and analyzed using standard methods.Results. The analysis of cytogenetic and karyological indicators of buccal epithelial cells in patients with CRAS revealed that, at the peak of the inflammatory process, cells with micronuclei and various forms of nuclear protrusions were more frequently observed. During therapy, there was a notable reduction in buccal epithelial cells exhibiting these pathological features.Conclusion. Before therapy, patients with RAS exhibited cytogenetic instability and various karyological anomalies in the buccal epithelial cells of the oral mucosa. These anomalies included significantly higher occurrences of micronuclei, karyopyknosis, karyolysis, nuclear vacuolization, intercellular bridges, and chromatin condensation. During the different therapy regimens for RAS, normalization of the epithelial state of the oral mucosa was observed, with the most pronounced improvements seen with the use of octenidine + phenoxyethanol in combination with a hydrogel-based material containing sodium alginate, Derinat, and lidocaine.

Comprehensive in vitro and in vivo evaluation of therapeutic potential of Bacopa-derived asiatic acid against a human oral pathogen Streptococcus mutans.

Dental caries is a common human oral disease worldwide, caused by an acid-producing bacteria Streptococcus mutans. The use of synthetic drugs and antibiotics to prevent dental caries has been increasing, but this can lead to severe side effects. To solve this issue, developing and developed countries have resorted to herbal medicines as an alternative to synthetic drugs for the treatment and prevention of dental caries. Therefore, there is an urgent need for plant-derived products to treat such diseases. Bacopa monnieri, a well-documented medicinal plant, contains 52 phytocompounds, including the pentacyclic triterpenoid metabolite known as asiatic acid (ASTA). Hence, this study aimed to demonstrate, for the first time, the antibacterial activity of phytocompound ASTA against S. mutans. The findings revealed that ASTA significantly inhibited the growth of S. mutans and the production of virulence factors such as acidurity, acidogenicity, and eDNA synthesis. Molecular docking analysis evaluated the potential activity of ASTA against S. mutans virulence genes, including VicR and GtfC. Furthermore, toxicity assessment of ASTA in human buccal epithelial cells was performed, and no morphological changes were observed. An in vivo analysis using Danio rerio (zebrafish) confirmed that the ASTA treatment significantly increased the survival rates of infected fish by hindering the intestinal colonization of S. mutans. Furthermore, the disease protection potential of ASTA against the pathognomonic symptom of S. mutans infection was proven by the histopathological examination of the gills, gut, and kidney. Overall, these findings suggest that ASTA may be a promising therapeutic and alternative drug for the treatment and prevention of oral infection imposed by S. mutans.