Buccal Cells Research Articles

Buccal Mesenchymal Stromal Cells as a Source of Osseointegration of Titanium Implants.

We studied the interaction of human buccal mesenchymal stem cells (MSCs) and osteoblasts differentiated from them with the surface of titanium samples. MSCs were isolated by enzymatic method from buccal fat pads. The obtained cell culture was presented by MSCs, which was confirmed by flow cytometry and differentiation into adipocytes and osteoblasts. Culturing of buccal MSCs on titanium samples was accompanied by an increase in the number of cells for 15 days and the formation of a developed network of F-actin fibers in the cells. The viability of buccal MSCs decreased by 8 days, but was restored by 15 days. Culturing of osteoblasts obtained as a result of buccal MSC differentiation on the surface of titanium samples was accompanied by a decrease in their viability and proliferation. Thus, MSCs from buccal fat pads can be used to coat implants to improve osseointegration during bone reconstruction in craniofacial surgery and dentistry. To improve the integration of osteoblasts, modification of the surface of titanium samples is required.

ГЕНЫ НЕЙРОМЕДИАТОРНЫХ СИСТЕМ МОЗГА, ДЕТЕРМИНИРУЮЩИЕ ПСИХОЭМОЦИОНАЛЬНЫЙ СТАТУС ЧЕЛОВЕКА

Background. In recent decades, there has been a significant interest worldwide in the clarification of the genetic mechanisms of the human psychoemotional status. The identification of genetic markers associated with psychoemotional status expands the possibilities of considering individual characteristics in career guidance, as well as in choosing methods of individual psychotherapy. Purpose. To identify the most informative polymorphic variants of genes associated with the human psychoemotional status. Material and methods. Psychological and molecular genetic testing was conducted among representatives of two groups of the Belarusian population: a control group (518 people) and a group of patients with personality disorders and deviant behavior (534 people). The following psychological testing methods were used: PSS-10 and PSS-14 – Russian-language versions of the Perceived Stress Scale-10, HADS-T – a test that measures symptoms of anxiety, and HADS-D – a test that measures symptoms of depression. DNA isolated from buccal epithelial cells was used as biological material for molecular genetic research. Genomic DNA was isolated using an extraction kit from Primetech LLC (Belarus). DNA concentration was assessed using a Qubit fluorimeter (Invitrogen, USA), as well as a Promega GloMax® Explorer Multimode Microplate Reader (USA). Results. A total of 33 polymorphic variants of genes of the brain neurotransmitter systems have been studied. 8 polymorphic variants were found to have statistically significant different frequencies in the studied groups, and 3 ones to have frequencies that differ at the level of tendency. Conclusions. The results obtained made it possible to form a panel of the most informative polymorphic variants of genes of the brain neurotransmitter systems, which can be used to determine individual psycho-emotional characteristics.

Local Low-dose Anti-PD-L1 Antibodies Improve Antitumor Effects in Oral Squamous Cell Carcinoma.

Immune checkpoint inhibitors are highly effective for treating recurrent and metastatic head and neck cancers. However, they require systemic administration and are associated with immune-related adverse events (irAEs). Reducing therapeutic antibody doses to prevent irAEs is challenging. Mouse buccal mucosa squamous cell carcinoma cells (Sq-1979) were transplanted into the backs of mice to induce tumors. The antitumor efficacy and tumor immunohistological environment in tumor-bearing mice were compared after administering a standard dose of programmed death-ligand 1 (PD-L1) antibodies systemically (200 mg/body) or 1/10th of the standard dose (20 mg/body) directly to tumors. Mice received four doses of antibody administered in 3-day intervals. Tumor reduction rates and antitumor efficacies were evaluated 21 days after initiating treatment. CD8+T cell counts and PD-L1, PD-1, perforin, and granzyme B levels; CD25 and Foxp3 expression levels; and tumor Tregs were assessed in the resected subcutaneous tumors. The antitumor efficacies in the local low-dose and systemic standard-dose groups were compared with that of the control group. The efficacies of the two treatment groups were similar, and both treatment groups revealed significant antitumor effects compared to the control group. Perforin and granzyme B levels were higher in the local low-dose group (p<0.05). Local low-dose administration of anti-PD-L1 antibodies exhibits antitumor efficacy similar to systemic standard-dose administration suggesting that local low-dose administration is useful for treating oral squamous cell carcinoma.

Human buccal epithelial cells as a model system for molecular analysis of DNA, RNA and protein

We report use of human buccal epithelial cells as an easy model system for isolation and molecular analysis of genomic DNA, RNA, and protein to study any gene of interest by Polymerase Chain Reaction (PCR), RNA expression by Reverse Transcription-PCR (RT-PCR), protein-profiling, and expression by western blot as well as DNA-methylation by Msp I/Hpa II-restriction digestion. We used simple methods to isolate genomic DNA, RNA and protein from human buccal cells collected by oral swab and cultured them in-vitro. The microscopic observation of haematoxylin and eosin (EA-50) stained cells, genomic PCR of house-keeping genes (GAPDH and β-actin), RT-PCR of GAPDH and β-actin mRNAs and whole cell protein-profiling by Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) were carried out. Expression of β-actin protein in supernatant and pellet fractions of the cells was determined by western blot analysis. MTT-assay was carried out to assess the cell viability and cell growth. Green Fluorescence Protein (GFP)-DNA was expressed in these cells by transient transfection. DNA-methylation in the genome was analyzed by Msp I/ Hpa II restriction digestion and agarose gel electrophoresis. Thus these methods can be used for molecular analysis of DNA, RNA and protein from the human buccal epithelial cells for studying and monitoring health, disease, population genetics/genomics and epidemiology under different conditions.

Вплив паління на цитогенетичний статус слизової оболонки ротової порожнини в осіб підліткового та юнацького віку

Cytogenetic monitoring provides information on both the health status of the population under study and the impact of genotoxic factors, i.e., the cytogenetic status of a person can serve as a biomarker of the level of exposure to genotoxicants, including various types of smoking. Purpose - to investigate the effect of different types of smoking on the cytogenetic status, namely, to analyse the cariological parameters of buccal epithelial cells of the oral mucosa in teenagers and young adults. Materials and methods. The cytogenetic status was studied by analysing the karyological parameters of buccal epithelial cells of the oral mucosa in 114 adolescents aged 15 to 24 years, who were divided into groups: the Group I included 26 people who smoke traditional cigarettes; the Group II - 22 people who smoke electronic cigarettes (vapes); the Group III - 23 people who smoke tobacco heating devices (IQOS); the Group IV - 43 people without smoking habits. Results. A significant increase in epithelial cells of buccal epithelium with nuclear disorders was found in smokers compared to the group of people without smoking habits. In addition, in individuals without smoking habits, no nuclear disorders such as protrusions, proliferation rates, early stage of nucleus destruction and completion of nucleus destruction were detected. Conclusions. The sensitivity of buccal epithelial cells to the effects of different types of smoking was established, which affects changes in cariological parameters. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the author.

Exploring the association between PITX2, third molars agenesis and sella turcica morphology

ObjectivesPITX2 is required for mammalian development and single nucleotide polymorphisms (SNPs) in this gene could be involved in dental agenesis and sella turcica patterns. Thus, the present study evaluated the association between SNPs in PITX2, third molars agenesis and sella turcica phenotypes.Materials and methodsThe sample consisted of healthy orthodontic German patients with lateral cephalometric radiographs with clearly visualization of the sella turcica, and dental orthopantomograms. The morphological variations of the sella turcica were evaluated using the lateral cephalograms, while third molar agenesis was evaluated using orthopantomograms. DNA isolated from buccal cells was used for genotyping three SNPs in PITX2 (rs3796902, rs1947187, and rs2595110). The analyzes were performed using a significance of 5%. There was no association between third molar agenesis and sella turcica phenotypes (p > 0.05). SNPs in PITX2 were also not associated with third molars agenesis (p > 0.05).ResultsSNPs in PITX2 were associated with sella turcica phenotypes. The rs3796902 was associated with hypertrophic posterior clinoid process (p = 0.013). The rs1947187 and rs2595110 were associated with sella turcica bridge type A (p = 0.013 and p = 0.011, respectively for genotype distribution). Patients that carry the genotypes GG-CC-AG (rs3796902- rs1947187- rs2595110) had 7.2 higher chance to present sella turcica bridge type A (p = 0.002; Odds ratio = 7.2, Confidence interval 95% 2.04–27.04).ConclusionsThird molar agenesis was not associated with SNPs in PITX2 and sella turcica phenotypes. SNPs in PITX2 may have an important role in sella turcica pattern.

Prognostic risk factors of buccal squamous cell carcinoma: A case-control study.

To describe the clinicopathologic presentation of buccal squamous cell carcinoma and identify risks factors for recurrence and overall survival. This is a retrospective case-control study of patients with oral cavity squamous cell carcinoma (OCSCC) treated at a single tertiary care center between 2010 and 2022. All patients with buccal subsite OCSCC treated during this time frame were included and paired with a randomly selected age and gender matched patient with non-buccal OCSCC. Relevant data was collected via chart review. Seventy-seven patients with buccal SCC were matched with 77 non-buccal OCSCC controls. The median follow-up time was 27 months (IQR 14-61). Median age was 67 years (IQR 57-75) and 53% of the cohort was female. Twenty (26%) buccal SCC patients experienced a recurrence versus 19 (25%) in the controls. Age ≥65-years-old increased odds of all-cause mortality in the buccal SCC group, but not in the control group. Perineural invasion and positive margins increased odds of recurrence in the buccal group only. Overall survival and progression-free survival did not differ between the groups, despite a greater number of T2 buccal tumors and T1 non-buccal tumors. Buccal SCC presents at a higher T stage than other oral cavity SCC subsite and may exhibit variance in the pathologic risk factors that predict poor outcomes versus non-buccal OCSCC. Despite these relatively minor differences, however, oncologic outcomes between these groups were similar.

Improvements in lung function following vitamin C supplementation to pregnant smokers are associated with buccal DNA methylation at 5 years of age

BackgroundWe previously reported in the “Vitamin C to Decrease the Effects of Smoking in Pregnancy on Infant Lung Function” randomized clinical trial (RCT) that vitamin C (500 mg/day) supplementation to pregnant smokers is associated with improved respiratory outcomes that persist through 5 years of age. The objective of this study was to assess whether buccal cell DNA methylation (DNAm), as a surrogate for airway epithelium, is associated with vitamin C supplementation, improved lung function, and decreased occurrence of wheeze.MethodsWe conducted epigenome-wide association studies (EWAS) using Infinium MethylationEPIC arrays and buccal DNAm from 158 subjects (80 placebo; 78 vitamin C) with pulmonary function testing (PFT) performed at the 5-year visit. EWAS were performed on (1) vitamin C treatment, (2) forced expiratory flow between 25 and 75% of expired volume (FEF25–75), and (3) offspring wheeze. Models were adjusted for sex, race, study site, gestational age at randomization (≤ OR > 18 weeks), proportion of epithelial cells, and latent covariates in addition to child length at PFT in EWAS for FEF25–75. We considered FDR p < 0.05 as genome-wide significant and nominal p < 0.001 as candidates for downstream analyses. Buccal DNAm measured in a subset of subjects at birth and near 1 year of age was used to determine whether DNAm signatures originated in utero, or emerged with age.ResultsVitamin C treatment was associated with 457 FDR significant (q < 0.05) differentially methylated CpGs (DMCs; 236 hypermethylated; 221 hypomethylated) and 53 differentially methylated regions (DMRs; 26 hyper; 27 hypo) at 5 years of age. FEF25–75 was associated with one FDR significant DMC (cg05814800), 1,468 candidate DMCs (p < 0.001), and 44 DMRs. Current wheeze was associated with 0 FDR-DMCs, 782 candidate DMCs, and 19 DMRs (p < 0.001). In 365/457 vitamin C FDR significant DMCs at 5 years of age, there was no significant interaction between time and treatment.ConclusionsVitamin C supplementation to pregnant smokers is associated with buccal DNA methylation in offspring at 5 years of age, and most methylation signatures appear to be persistent from the prenatal period. Buccal methylation at 5 years was also associated with current lung function and occurrence of wheeze, and these functionally associated loci are enriched for vitamin C associated loci.Clinical trial registration ClinicalTrials.gov, NCT01723696 and NCT03203603.

Cross-tissue comparison of telomere length and quality metrics of DNA among individuals aged 8 to 70 years.

Telomere length (TL) is an important biomarker of cellular aging, yet its links with health outcomes may be complicated by use of different tissues. We evaluated within- and between-individual variability in TL and quality metrics of DNA across five tissues using a cross-sectional dataset ranging from 8 to 70 years (N = 197). DNA was extracted from all tissue cells using the Gentra Puregene DNA Extraction Kit. Absolute TL (aTL) in kilobase pairs was measured in buccal epithelial cells, saliva, dried blood spots (DBS), buffy coat, and peripheral blood mononuclear cells (PBMCs) using qPCR. aTL significantly shortened with age for all tissues except saliva and buffy coat, although buffy coat was available for a restricted age range (8 to 15 years). aTL did not significantly differ across blood-based tissues (DBS, buffy coat, PBMC), which had significantly longer aTL than buccal cells and saliva. Additionally, aTL was significantly correlated for the majority of tissue pairs, with partial Spearman's correlations controlling for age and sex ranging from ⍴ = 0.18 to 0.51. We also measured quality metrics of DNA including integrity, purity, and quantity of extracted DNA from all tissues and explored whether controlling for DNA metrics improved predictions of aTL. We found significant tissue variation: DNA from blood-based tissues had high DNA integrity, more acceptable A260/280 and A260/230 values, and greater extracted DNA concentrations compared to buccal cells and saliva. Longer aTL was associated with lower DNA integrity, higher extracted DNA concentrations, and higher A260/230, particularly for saliva. Model comparisons suggested that incorporation of quality DNA metrics improves models of TL, although relevant metrics vary by tissue. These findings highlight the merits of using blood-based tissues and suggest that incorporation of quality DNA metrics as control variables in population-based studies can improve TL predictions, especially for more variable tissues like buccal and saliva.

Up-regulation of extracellular-matrix and inflammation related genes in oral squamous cell carcinoma

ObjectiveOral squamous cell carcinoma (OSCC) is the most prevalent malignancy with late-presentation, site-specific heterogeneity, and high-propensity for recurrence/metastasis that has shown rise in mortality. Lately, research emphasize on dynamic interactions between tumor-cells and extracellular-matrix components within tumor-microenvironment that promote tissue integrity loss and carcinogenesis. Therefore, OSCC clinical-management is still challenging. DesignPresent study validated clinical utility of a 13 gene-panel in two chief sub-sites of OSCC: Buccal mucosa squamous cell carcinoma (BMSCC) (N = 50) and Tongue squamous cell carcinoma (TSCC) (N = 52) using qRT-PCR. Principal component analysis and binary logistic regression analysis were applied to acquire definite multi gene models. Protein expression analysis was employed using the Human Protein Atlas, UALCAN and TIMER 2.0 databases to explore potential correlation between immune cells and gene-panels. ResultsSignificant up-regulation of CXCL8, CXCL10, FN1, GBP1, IFIT3, ISG15, MMP1, MMP3, MMP10, PLAU, SERPINE1 and SPP1 except OASL was observed in OSCC tissue in comparison of absolute normal controls. Although, this gene-panel could potentially discriminate OSCC tissues from absolute normal controls as solitarily diagnostic and/or predictive biomarkers, models generated also showed substantial discriminating efficacy. Eight-genes were found to be significantly associated with poor-prognosis on clinico-pathological association. Protein-expression confirmed overexpression of gene-panel and added advantage of being secretory-protein. Importantly, up-regulated genes in our study showed significant relation with immune-cells infiltration suggesting their contribution in immune-escape. ConclusionThus, we propose that the 13 gene-panel could pave the way to effective and personalized clinical-management of OSCC in terms of diagnostic and prognostic measures and thereby as therapeutic targets.

Antifibrotic effect of silymarin on arecoline-induced fibrosis in primary human buccal fibroblasts: an in silico and in vitro analysis.

This study aimed to assess silymarin's anticancer and antifibrotic potential through in silico analysis and investigate its impact on in vitro arecoline-induced fibrosis in primary human buccal fibroblasts (HBF). The study utilized iGEMDOCK for molecular docking, evaluating nine bioflavonoids, and identified silymarin and baicalein as the top two compounds with the highest target affinity, followed by subsequent validation through a 100ns Molecular Dynamic Simulation demonstrating silymarin's stable behavior with Transforming Growth Factor Beta. HBF cell lines were developed from tissue samples obtained from patients undergoing third molar extraction. Arecoline, a known etiological factor in oral submucous fibrosis (OSMF), was employed to induce fibrogenesis in these HBFs. The inhibitory concentration (IC50) of arecoline was determined using the MTT assay, revealing dose-dependent cytotoxicity of HBFs to arecoline, with notable cytotoxicity observed at concentrations exceeding 50µM. Subsequently, the cytotoxicity of silymarin was assessed at 24 and 72 h, spanning concentrations from 5µM to 200µM, and an IC50 value of 143µM was determined. Real-time polymerase chain reaction (qPCR) was used to analyze the significant downregulation of key markers including collagen, epithelial-mesenchymal transition (EMT), stem cell, hypoxia, angiogenesis and stress markers in silymarin-treated arecoline-induced primary buccal fibroblast cells. Silymarin effectively inhibited fibroblast proliferation and downregulated genes associated with cancer progression and EMT pathway, both of which are implicated in malignant transformation. To our knowledge, this study represents the first exploration of silymarin's potential as a novel therapeutic agent in an in vitro model of OSMF.

Mitochondrial proteins as biomarkers of occupational disease risk of pilots and astronauts

Introduction. Mitochondrial dysfunction is an important pathogenic mechanism of neurodegeneration, characterized by a progressive structural and functional loss of neurons, leading to heterogeneous clinical and pathological manifestations with subsequent impairment of the functional anatomy of the brain. Aim of research. To study the influence of occupational hazards and stress experienced by civil aviation pilots and cosmonauts on the expression of mitochondrial biomarkers in buccal epithelial cells to assess the risk of developing neurodegenerative processes. Material and methods. The study involved 23 male participants in two age groups. 4 groups of investgation were formed, according to the occupation, comparable in age. The expression of mitochondrial proteins prohibitin and parkin in the buccal epithelium of the study participants was assessed by immunohistochemical methods. Results. A decrease in the expression level of the prohibitin protein was found in the group of civil aviation pilots compared to the control group of the corresponding age. There was also a tendency to a decrease in the level of expression of the studied proteins prohibitin and parkin in the group of cosmonauts compared with the control group of the corresponding age. Conclusion. The results obtained indicate a mitochondrial dysfunction, which may increase the risk of developing neurodegenerative changes.

New insights into the genetics of mandibular retrognathism: novel candidate genes.

Mandibular retrognathism (MR) is acommon skeletal malocclusion in humans with astrong genetic component. Single nucleotide polymorphisms (SNPs) in genes encoding epidermal growth factor (EGF) and EGF receptor (EGFR) could be involved in the etiology of mandibular retrognathism. Therefore, in this study, we investigated whether SNPs in the genes encoding for EGF and EGFR are associated with MR in German teenagers. This nested case-control study evaluated German orthodontic patients, aged 10-18years. DNA, which was isolated from buccal epithelial cells using two cytobrushes, was used for genotyping analysis and digital pretreatment lateral cephalograms were examined to calculate SNB and ANB. Patients with aretrognathic mandible (SNB < 78°) were included as cases, while patients with an orthognathic mandible (SNB = 78-82°) were included as controls. Four SNPs in the genes encoding for EGF and EGFR were chosen and genotyped using real-time PCR. Allele, genotype, and haplotype frequency were compared across groups (α = 5%). Finally, 119 patients were included in this study (45orthognathic mandible, 74retrognathic mandible). The minor alleleG in rs4444903 (EGF) was statistically more frequent in individuals with an orthognathic mandible (p = 0.008). The haplotype formed by the mutant alleles for rs4444903|rs2237051 (EGF; G|A) was statistically more frequent in the orthognathic mandible group (p = 0.007). The SNPs rs4444903 and rs2237051 in EGF, and rs2227983 in EGFR were statistically associated with adecreasing risk of developing aretrognathic mandible according to univariate and multivariate statistical analysis (p < 0.05). SNPs in EGF (rs4444903 and rs2237051) and EGFR (rs2227983) were associated with MR in our German sample and could be genetic biomarkers for early and individualized diagnostic identification of retrognathic mandibular development by means of genetic screening tests.

KARYOLOGICAL CHANGES OF BUCCAL EPITHELIOCYTES IN CHILDREN WITH ACUTE RESPIRATORY PATHOLOGY CAUSED BY SARS-COV-2

Pneumonia is one of the leading causes of death in children under 5 years of age in lowincome countries worldwide. This problem became especially acute with the onset ofthe new coronaviral infection caused by SARS-CoV-2. Community- acquired pneumoniais also one of the most common diseases in children, which requires extensive use ofantibiotics and admission to pediatric clinics. That is why diagnosis of the infectiousinflammatory process activity and prediction of the disease severity is the main factorin optimizing the treatment of children who suffer from infectious respiratory pathology.Objective – to improve the prediction of the severe course of acute respiratory pathologycaused by the SARS-CoV-2 virus by analyzing the results of kariological indicators ofexfoliative cells of the buccal epithelium in children of different ages. Materials and methods. 63 children with acute respiratory pathology caused by theSARS-CoV-2 virus (average age – 10.6±0.5 years) were comprehensively examinedby the method of simple random sampling using the «experiment- control» method.The verification of the etiological factor of inflammation of the respiratory tract wascarried out by the method of polymerase chain reaction with reverse transcriptase(a set of COVID-19 Multiplex RT-PCR in real time from Labsystems Diagnostics Oy) ina nasopharyngeal swab and/or lung expiratory tissue.Fixed smears- imprints of exfoliative cells of the buccal epithelium were stained with azureeosin according to Romanovsky- Giemsa. By light microscopy of cytological samples,changed cells with nuclear abnormalities were evaluated and the number of cells withmicronuclei and karyological indicators of exfoliative cells of the buccal epithelium per100 cells in the preparation were determined.Results.Pathological changes of buccal epithelium exfoliative cells as protrusion, karyopyknosis,karyorrhexis and apoptosis are directly proportional to the disease severity and reflect exclusivelytoxic effects. Identification of the morphological changes expressiveness in the buccal epitheliumcells can be used as a diagnostic factor in prediction of the severe disease, and accordingly,serves as an impetus in optimization of the conducted treatment strategy.Conclusions. A greater number of pathological cells in smears- imprints of the buccalepithelium, and, accordingly, slightly higher cytogenetic indices are determined in patientswith pneumonia and multisystem inflammatory syndrome associated with SARS-CoV-2, whohad a severe and extremely severe course of the disease. Significantly fewer pathologicallychanged cells are found in patients with acute respiratory tract infection of multiplelocalization and bronchitis, who mostly had a medium- severe course of the disease, whichcan be used to predict the severity of the infectious- inflammatory process in children.

Photobiomodulation Effect of Different Diode Wavelengths on the Proliferation of Human Buccal Fat Pad Mesenchymal Cells

This study aimed to determine the most effective wavelength for the proliferation of Human Buccal Fat Pad Mesenchymal Stem Cells (BFPMSCs) in cell culture. These cells can be used for different purposes such as regenerative periodontal procedures. Materials and Methods: The wells containing BFPMSCs were subjected to laser irradiation at 635, 660, 808, and 980 nm wavelengths with 1, 1.5, 2.5, and 4 J/cm2 energy densities. Cell proliferation and viability were evaluated after 1, 3, and 5 days with the methyl thiazolyl tetrazolium (MTT) assay. Result: The proliferation rate of human Buccal Fat Pad Mesenchymal Cells (BFPMSCs) was increased on the first and third days at a wavelength of 808 nm and day five at a wavelength of 980 nm in comparison to the control group. Our findings distinguished that PBMT with 635, 660, 808, and 980 nm wavelengths increased the proliferation of BFPMSCs. Conclusion: The best laser radiation setting, which led to the highest proliferation rate of the cells, included a wavelength of 808 nm with 2.5 J/cm2 energy density.

Cytogenetic disorders in cells of mucous membranes under the influence of welding aerosols

Preserving the health of the working population is a priority direction of state policy. At present, the search for the most informative physiological, biochemical, immunological and other criteria for assessing the action of harmful factors is of great importance. The purpose of the study is to study the cytological structures in the cells of the mucous membranes when exposed to welding aerosols on the body of electric and gas welders. A diagnostic examination was carried out for electric and gas welders of the metallurgical plant of the Republic of Bashkortostan. Cytological laboratory parameters of cells of the mucous membranes of the oral cavity (buccal epithelium) and discharge from the nose (rhinocytogram) were analyzed under the influence of a harmful adverse production factor — welding aerosol (SA). The index of accumulation of cytogenetic disorders was used in the work — Index of accumulation of cytogenetic damage (Iac), which takes into account the indicators of cellular kinetics. The general hygienic assessment of the working conditions of electric and gas welders according to the criteria R 2.2.2006–05 corresponds to the harmful 3rd class of 2–3 degrees (3.1–3.2). The results of the study revealed cytogenetic disorders of buccal epithelium cells in 95,65 % of electric and gas welders. A low probability of cytogenetic disorders was found in 24,64 %, moderate — in 56,52 %, high — in 18,84 % of workers. When evaluating the rhinocytogram, an increase in the number of segmented neutrophils was found in 86,96 %, eosinophils in 39,13 %, reflecting nonspecific (microbial) and allergic inflammation. The data obtained made it possible to identify the important role of diagnostic procedures for identifying indicators of health disorders among the working population and the development of individual preventive measures.

Evaluating the effect of strontium ranelate and photobiomodulation on cementogenic and osteogenic differentiation of buccal fat pad-derived stem cells: An in vitro study.

This study aimed to analyze the impact of strontium ranelate (Str), photobiomodulation (PBM), or their combination of the proliferation, osteogenic differentiation, and cementogenic differentiation of buccal fat pad-derived stem cells. BFPdSCs were exposed to one of the following interventions: (1) PBM (660 nm), (2) PBM (660 nm) + Str, (3) PBM (880 nm), (4) PBM (880 nm) + Str, (5) Str. All study groups had significantly higher osteogenic differentiation than the control group (p < 0.05), and no significant difference existed between the 660 and 808 nm groups (p = 0.97). Compared to the Str group, 660 nm and 880 nm group samples had significantly lower osteogenic differentiation (p < 0.0001), while other groups did not show a significant difference. Regarding cementogenic differentiation, the 660 nm group showed higher values than the 808 nm group (p < 0.01). Compared with the Str group, 660 nm, 660 nm + Str, and 808 nm + Str groups showed significantly higher gene expression (p < 0.05). In the case of osteogenic differentiation, although photobiomodulation alone had a lower inducing effect than strontium ranelate, combining 808 nm diode lasers and strontium ranelate may provide the best results. Moreover, using a 660 nm diode laser and exposing stem cells to strontium ranelate can be the most effective approach to induce cementogenic differentiation.

3D bioprinted small extracellular vesicles from periodontal cells enhance mesenchymal stromal cell function

Recent research indicates that combining 3D bioprinting and small extracellular vesicles (sEVs) offers a promising ‘cell-free’ regenerative medicine approach for various tissue engineering applications. Nonetheless, the majority of existing research has focused on bioprinting of sEVs sourced from cell lines. There remains a notable gap in research regarding the bioprinting of sEVs derived from primary human periodontal cells and their potential impact on ligamentous and osteogenic differentiation. Here, we investigated the effect of 3D bioprinted periodontal cell sEVs constructs on the differentiation potential of human buccal fat pad-derived mesenchymal stromal cells (hBFP-MSCs). Periodontal cell-derived sEVs were enriched by size exclusion chromatography (SEC) with particle-shaped morphology, and characterized by being smaller than 200 nm in size and CD9/CD63/CD81 positive, from primary human periodontal ligament cells (hPDLCs) and human gingival fibroblasts (hGFs). The sEVs were then 3D bioprinted in 10 % gelatin methacryloyl (GelMA) via microextrusion bioprinting. Release of sEVs from bioprinted constructs was determined by DiO-labelling and confocal imaging, and CD9 ELISA. Attachment and ligament/osteogenic/cementogenic differentiation of hBFP-MSCs was assessed on bioprinted GelMA, without and with sEVs (GelMA/hPDLCs-sEVs and GelMA/hGFs-sEVs), scaffolds. hBFP-MSCs seeded on the bioprinted sEVs constructs spread well with significantly enhanced focal adhesion, mechanotransduction associated gene expression, and ligament and osteogenesis/cementogenesis differentiation markers in GelMA/hPDLCs-sEVs, compared to GelMA/hGFs-sEVs and GelMA groups. A 2-week osteogenic and ligamentous differentiation showed enhanced ALP staining, calcium formation and toluidine blue stained cells in hBFP-MSCs on bioprinted GelMA/hPDLCs-sEVs constructs compared to the other two groups. The proof-of-concept data from this study supports the notion that 3D bioprinted GelMA/hPDLCs-sEVs scaffolds promote cell attachment, as well as ligamentous, osteogenic and cementogenic differentiation, of hBFP-MSCs in vitro.

Evaluation of Cytotoxic and Genotoxic Effects in Buccal Mucosal Cells in Non-Smokers and Users of Traditional Combustible Tobacco Products and Non-Combustible Alternatives.

The aim of this cross-sectional observational study was to investigate cytogenetic damage to the buccal mucosa in non-smokers and consumers of traditional combustible tobacco products and non-combustible alternatives. A total of 160 participants were divided into four groups according to the type of product used, including non-smokers, users of conventional combustible tobacco (cigarettes), heated tobacco, and electronic, tobacco-free vapor products (e-cigarettes). Buccal mucosa samples were analyzed using the micronucleus cytome assay to assess cytotoxic and genotoxic damage. E-cigarette users showed significantly higher values for all tested parameters in the micronucleus test compared to non-smokers (p < 0.05). Similarly, users of tobacco heating products showed an increase in all parameters (p < 0.05), with the exception of the number of cells with micronuclei. Conventional cigarette smokers showed a notable increase in the number of binucleated cells and cells with karyorrhexis and karyolysis (p ≤ 0.05). When assessing the differences between users of traditional combustible tobacco products and non-combustible alternatives, these did not appear to be significant, except for e-cigarette users, who had significantly more cells with condensed chromatin (p ≤ 0.001), while users of tobacco heating products had more pyknotic cells (p ≤ 0.001). The results of this study underscore the heightened occurrence of cytotoxic and genotoxic damage in users of both conventional combustible tobacco products and non-combustible alternatives compared to non-smokers, emphasizing the detrimental impact of these products on the oral mucosa.

RFX transcription factor in the human-associated yeast Candida albicans regulates adhesion to oral epithelium.

Adhesion to mucosal surfaces is a critical step in many bacterial and fungal infections. Here, using a mouse model of oral infection by the human fungal pathobiont Candida albicans, we report the identification of a novel regulator of C. albicans adhesion to the oral mucosa. The regulator is a member of the regulatory factor X (RFX) family of transcription factors, which control cellular processes ranging from genome integrity in model yeasts to tissue differentiation in vertebrates. Mice infected with the C. albicans rfx1 deletion mutant displayed increased fungal burden in tongues compared to animals infected with the reference strain. High-resolution imaging revealed RFX1 transcripts being expressed by C. albicans cells during infection. Concomitant with the increase in fungal burden, the rfx1 mutant elicited an enhanced innate immune response. Transcriptome analyses uncovered HWP1, a gene encoding an adhesion protein that mediates covalent attachment to buccal cells, as a major RFX1-regulated locus. Consistent with this result, we establish that C. albicans adhesion to oral cells is modulated by RFX1 in an HWP1-dependent manner. Our findings expand the repertoire of biological processes controlled by the RFX family and illustrate a mechanism whereby C. albicans can adjust adhesion to the oral epithelium.