Buccal Epithelial Cells Research Articles

Sodium glycodeoxycholate and sodium deoxycholate as epithelial permeation enhancers: in vitro and ex vivo intestinal and buccal bioassays

Bile salts were first tested as epithelial permeation enhancers (PEs) for the intestine and buccal routes over 20 years ago. They are not as popular as other PEs due to their non-specific mechanism of action and perceived toxicity potential. We revisited two of them by comparing efficacy and toxicity of sodium glycodeoxycholate (SGC) and sodium deoxycholate (DC) for both routes using in vitro and ex vivo methods. Cytotoxicity assays in Caco-2 cells revealed that both agents altered cellular parameters at concentrations >2 mM over 60 min. Both agents reduced the transepithelial resistance (TEER) and doubled the Papp of [3H]-octreotide across isolated rat colonic mucosae mounted in Ussing chambers at 10 mM concentrations. In some studies, 10 mM GDC also increased the Papp of the paracellular marker, FITC-dextran 4000 (FD4) and the fluorescent peptide, FITC-LKP, across colonic mucosae. Tissue histology was intact despite some mild perturbation at 10 mM. In the buccal epithelial cell line, TR146, changes in cell parameters were also seen at 1.5 mM over 60 min for both agents, with slightly more sensitivity seen for DC. In isolated porcine buccal epithelial mucosae, GDC was slightly more potent and efficacious than DC at increasing the Papp of [14C]-mannitol. It also increased the Papp of [3H]-octreotide and FITC-LKP by ∼3-fold across porcine buccal tissue without causing damage. Overall, GDC and DC were efficacious in intestinal and buccal models. Both cause mild perturbation leading to an increase in paracellular fluxes for hydrophilic molecules including peptides. Their moderate efficacy, low potency, and low toxicity in these models are similar to that of more established PEs in clinical trials.

A Disorder of Sex Development in a Holstein-Friesian Heifer with a Rare Mosaicism (60,XX/90,XXY): A Genetic, Anatomical, and Histological Study.

Simple SummaryDisorders of sex development (DSDs) are congenital conditions in which a discordance between chromosomal, gonadal, or phenotypic sex is observed. DSDs are serious problems in animal breeding, as they lead to sterility. In cattle, the most common form of DSD is freemartinism, which manifests as the presence of leukocyte chimerism (XX/XY), and occurs in heifers originating from heterosexual twin pregnancy. Other forms of DSD are rarely observed in this species. In this study, we describe a very rare diploid/triploid (60,XX/90,XXY) condition in a DSD heifer. Comprehensive clinical, anatomical, histopathological and genetic analysis was performed.In this study, we describe an eighteen-month-old Holstein–Friesian heifer with a deformed vulva, located abdominally. The heifer showed typical signs of estrus. A comprehensive anatomical and histopathological examination revealed a blind-ended vagina and an additional section of urethra, which became a part of the shortened penis. Cytogenetic analysis showed the presence of two cell lines: 60,XX and 90,XXY. The frequency of the triploid cell line was low (3%) in leukocytes and elevated (35%) in fibroblasts. The molecular detection of Y-linked genes (SRY and AMELY) in the blood, skin, hair follicles, and buccal epithelial cells confirmed the presence of a cell line carrying the Y chromosome. Genotyping of 16 microsatellite markers in DNA isolated from hair follicles and fibroblast culture showed the presence of one (homozygous) or two variants (heterozygous) at all the studied loci, and allowed chimerism to be excluded. We concluded that the heifer had diploid/triploid (60,XX/90,XXY) mosaicism. To our knowledge, this is only the fifth such case to be reported worldwide in this species. Since cytogenetic studies are routinely performed on in vitro cultured leukocytes, we suspect that the prevalence of this chromosome abnormality is underestimated, as it is known from published reports that the frequency of the triploid cell line is usually very low in leukocytes.

Internalizing symptoms associate with the pace of epigenetic aging in childhood

Childhood psychiatric symptoms may be associated with advanced biological aging. This study examined whether epigenetic age acceleration (EAA) associates with internalizing and externalizing symptoms that were prospectively collected across childhood in a longitudinal cohort study. At age 6 buccal epithelial cells from 148 children (69 girls) were collected to survey genome-wide DNA methylation. EAA was estimated using the Horvath clock. Internalizing symptoms at ages 2.5 and 4 years significantly predicted higher EAA at age 6, which in turn was significantly associated with internalizing symptoms at ages 6–10 years. Similar trends for externalizing symptoms did not reach statistical significance. These findings indicate advanced biological aging in relation to child mental health and may help better identify those at risk for lasting impairments associated with internalizing disorders.

Usnic acid deteriorates acidogenicity, acidurance and glucose metabolism of Streptococcus mutans through downregulation of two-component signal transduction systems

The principal etiological agent of human dental caries, Streptococcus mutans is a multi-virulent pathogen that can transform commensal oral microbial community to plaque biofilms. Major virulence factors that are associated with the cariogenicity of S. mutans include adhesion, acidogenicity and acidurity. All these pathogenic traits coordinate and alter the dental plaque ecology which provide room for interaction with other similar acidogenic and aciduric bacteria. This cariogenic flora increases the possibility of enamel demineralization which headway to caries development. The present study was aimed at evaluating the antimicrobial and antiinfective potential of a lichen secondary metabolite usnic acid (UA) against S. mutans. Minimum inhibitory concentration (MIC), Minimum bactericidal concentration (MBC) and growth kinetics were evaluated to determine the antimicrobial potential of UA against S. mutans. UA at 5 µg mL−1 and 10 µg mL−1 concentration were considered as MIC and MBC respectively. Effect on biofilm formation was microscopically assessed and found to be reduced in a concentration dependent manner. Gene expression of gtfB, gtfC, gtfD, vicR, ComDE and smu0630 was found to be downregulated upon treatment with sub-MIC of UA. Acidogenicity, acidurity, eDNA synthesis and response to oxidative stress were found to be attenuated by the influence of UA. It was also demonstrated to act on preformed mature biofilm of S. mutans. Moreover, UA was shown to possess very low frequency to acquire spontaneous resistance development in S. mutans. Besides, no morphological aberrations or toxic effect was instigated by UA in the human buccal epithelial cells as well as to the oral commensals. Altogether, these results demonstrate the therapeutic potential of usnic acid in the treatment of S. mutans infection.

Epicatechin and quercetin exhibit in vitro antioxidant effect, improve biochemical parameters related to metabolic syndrome, and decrease cellular genotoxicity in humans

Metabolic syndrome is a condition whose incidence has been increasing around the world. It promotes a metabolic state of chronic systemic inflammation, correlated to cellular stress and genetic mutations, and subsequently with deadly chronic diseases, such as type 2 diabetes mellitus, cardiovascular diseases, and cancer. A randomized placebo-controlled study (n = 156) was conducted to determine the effects of consuming an enriched bread with 0.05% of a 1:1 mixture of (−)-epicatechin and quercetin on anthropometric and biochemical parameters of the participants. As a result, total cholesterol, LDL-cholesterol, total triglycerides, and fasting plasma glucose significantly decreased after three months of daily enriched bread consumption. Nuclear abnormalities in buccal epithelium cells also decreased (15.8 ± 3.2 down to 8.3 ± 1.0), showing a genoprotective effect. The antioxidant properties of these compounds were observed by monitoring changes in the cytoplasmic redox tone of intact Caco-2 cells expressing HyPer, a fluorescent redox biosensor. The combination of (–)-epicatechin and quercetin changes the cytoplasmic redox ambient in living cells and significantly improves biochemical parameters related to metabolic syndrome, and decreases the number of cell abnormalities in buccal epithelium cells of patients.

DBD-FISH, an effective marker for detecting genotoxicity in buccal mucosa exfoliated cells of patients with oral cancer

Oral squamous cell carcinoma (OSCC) is characterized by increased genetic instability as an essential variable of event of neoplastic transformation. The aim of this study was to evaluate genomic instability in exfoliated cells from the buccal mucosa of patients with OSCC vs. the control group, using DNA Breakage Detection/Fluorescence In Situ hybridization (DBD-FISH). Exfoliated cells from the buccal mucosa were obtained from 38 patients with oral cancer (case group) and from 10 individuals without oral lesions (control group). DNA damage was evaluated by DBD-FISH using the whole-genome DNA probe and digital imaging analysis. Collaterally, HPV infection was determined utilizing the INNO-LiPA HPV kit. Patients with OSCC showed an increase in the hybridization signal five times more intense than that of the baseline level of DNA damage detected in control individuals. The best cutoff value for predicting oral squamous cell carcinoma was 67.46, and an Odds Ratio (OR) value of 87. HPV detection analysis revealed than one patient with OSCC (2.6%) was positive for HPV. All controls were negative HPV. In conclusion, DBD-FISH permitted the clear visualization of level high of DNA damage in the buccal epithelial cells of patients with OSSC respect to control group. Chromosome instability in oral mucosa may be an individual marker of malignant transformation in OSCC.

Experimental application of tissue engineered constructions for urethroplasty

Introduction. For diseases such as stricture and obliteration of the urethra, urethral hypospadias, reconstructive operations are required. The buccal mucosa is the material most commonly used for these operations. The search for alternative materials, carried out in order to reduce trauma and complications in the donor area, is an urgent area of modern urology. Tissue engineered constructions (TEC) can be used as such material.Objective. Justification of the possibility of applying a TEC based on the biodegradable polymers and seeded with autologous buccal epithelium (BE) cells as an implantable material for urethroplasty in an experiment.Methods and Materials. TEC based on poly-L-lactide-caprolactone (PLC) and poly-L-lactide-glycolide (PLG) seeded with BCs was created. Rabbits (n=12) underwent a biopsy of the oral mucosa, BCs were isolated, cultured and PLC-PLG scaffold was seeded with cells. TECs seeded with autological BCs were used on the model of acute trauma of rabbit urethra for replacement urethroplasty.Results. The results were evaluated after 12 weeks, according to the histological examination, there was a repair of the urethral mucosa. According to the data of retrograde urethrography, no impaired urethra patency was detected.Conclusion. TEC (PLK-PLG) seeded with autologous BCs ensured the maintenance of the rabbit urethral lumen which is necessary for adequate urination. This TEC could be recommended for the further clinical studies.

Взаимосвязь цитологических особенностей буккального эпителия и биохимических параметров у школьников с ожирением

Objective. To determine the relationship between metabolic parameters and cytological characteristics of the buccal mucosa in obese children. Patients and methods. An open cross-sectional case-control study included 78 schoolchildren aged 9 to 15 years, 44 of them were obese. We determined body mass index, lipid metabolism parameters (total cholesterol, high- and low-density lipoprotein cholesterol, apolipoproteins), and also cytological characteristics of buccal epithelial cells. Results. The number of buccal epithelial cells with condensed chromatin was by 10 times higher in obese children. A correlation was found between cholesterol concentrations and certain cytological parameters, but this correlation was statistically significant only in the pair “cholesterol – number of binucleated cells” (correlation coefficient 0.925, p = 0.05) in obese children and in the pair “total protein – binucleated cells” in practically healthy children. Conclusion. We confirmed the known data about specific characteristics of lipid metabolism in obese children that correlate with certain cytological parameters of the buccal mucosa, namely, an increased number of cells with chromatin condensation. This phenomenon is suggestive of activation of apoptosis in obese children. Keywords: buccal epithelial cells, obesity, schoolchildren, cholesterol

Age and AgNor-A morphometric study

To investigate and correlate total AgNOR area/Total nucleus area (TAA/TNA) values in buccal epithelial cells of healthy individuals in different age groups. In present study 50 healthy individuals are included with age ranging 10 to 60 years. These are divided into 5 groups. Group 1-10-20 year, Group2-20- 30year, Group3-30- 40year, Group 4-40-50-year, Group 5-50-60 year. Oral epithelial cells collected with the help of cyto brush from buccal mucosa. Smears were prepared on clean glass slides and fixed with 95% alcohol. Fixed slides Smears were stained with AgNOR stain. Statistically significant correlations were found between mean TAA/TNA values and age p < 0.001 for linear and p < 0.0001 for polynominal regression, and between AgNOR number and age, p < 0.001 for linear. There is a significant correlation between age and AgNOR amount (ribosome biosynthesis rate) in buccal epithelial cells of healthy individuals. AgNORs in buccal epithelial cells may be used for detection of age

Ассоциация относительной длины теломер и генетического варианта генa SIRT1 с возрастной макулярной дегенерацией

One of the most important factors predisposing to the development of age-related macular degeneration (AMD) is aging. Telomeres are important for aging by maintaining genome stability. Aim: to identify the association between relative telomere length of buccal epithelial cells and SIRT1 rs12778366 genetic variation and late AMD. Patients and Methods: 100 patients (200 eyes) were enrolled, i.e., 50 patients with AMD (AREDS category 4) and 50 patients without AMD. Genomic DNA isolated from buccal epithelial cells by phenol-chloroform extraction was used. Genotyping of SIRT1 rs12778366 polymorphic locus was performed by TaqMan® real-time PCR. Telomere length was measured by real-time PCR as described earlier [Cawthon, 2002] using specific primers. Relative telomere length was assessed by the relative telomere to single-copy gene (T/S) ratio. Results: the rate of allele C was 25% in the study group and 14% in the control group (p=0.049). The rate of heterozygotic TC genotype was twice higher in the study group compared to the control group (p=0.045). In heterozygotic carriers of the allele C of the SIRT1 rs12778366 gene, the risk of AMD is 2.048- and 2.425-times higher in сodominant and dominant inheritance pattern, respectively. In patients with late AMD, there are more short telomeres (64% vs. 48% in the control group, р=0.0002). Conclusions: further studies of a polymorphic SIRT1 gene locus in the association with telomere length in a larger sample are required. In the future, these molecular markers can be applied to predict the individual course of AMD and to implement preventive measures. Keywords: age-related macular degeneration, relative telomere length, rs12778366, SIRT1 gene, age-related diseases, buccal epithelium, genetic testing. For citation: Moshetova L.K., Dmitrenko O.P., Abramova O.I. et al. Association between relative telomere length and a genetic variant of SIRT gene and age-related macular degeneration. Russian Journal of Clinical Ophthalmology. 2021;21(3):143–146 (in Russ.). DOI: 10.32364/2311- 7729-2021-21-3-143-146.

Is duration of exposure a determinant factor for genotoxicity and clinical manifestations induced by Formaldehyde?

Introduction: Formaldehyde is a naturally occurring compound extensively used as cadavers' preservative in medical schools anatomy departments. The International Agency for Research on Cancer (IARC) classifies formaldehyde as carcinogenic to humans (group 1). Aim of the work: To examine duration of exposure as a determining factor in genotoxicity of formaldehyde exposure using the micronucleus test, correlate clinical manifestations with genotoxic effects, and measure formaldehyde air levels in dissecting rooms of Alexandria Faculty of Medicine, Egypt. Materials and methods: A pre-designed questionnaire was filled for each subject to assess clinical signs and symptoms of exposure and to screen for exclusion criteria. Buccal epithelial cells were collected; twice from 40 undergraduate students before and after one month of exposure, and once from 10 staff members who had participated in anatomy classes. Cells of both groups were stained using Feulgen method and micronucleus test (MN) was applied. The air levels were measured in 3 anatomy labs using MIRAN-IR. Results: Mean frequency of micronuclei was significantly higher in staff group (2.60 ±1.26)/1000 cells than the control and student groups (0.80±0.76)/1000 cells, (1.05 ±1.28)/1000 cells respectively with p= 0.001. Significant positive correlation between frequency of MN and skin, eye and respiratory symptoms. The mean level of airborne formaldehyde in 3 dissecting rooms was 7.2 ±6.9 ppm which is above OSHA recommendations. Conclusion: Duration of exposure is an important determinant in genotoxiciy of formaldehyde exposure. Clinical manifestations of formaldehyde exposure were correlated to the MN frequency. Studied participants are exposed to high concentrations of formaldehyde above the standard levels according to time of exposure. Efforts have to be made to improve air quality and reduce exposures during anatomy classes.

An exfoliative cytology study to assess oral mucosa of oral submucous fibrosis patients in Chennai, Tamil Nadu: A preliminary study

Background: Oral submucous fibrosis (OSMF) is a chronic, insidious disease that affects the oral mucosa. It is a progressive high risk potentially malignant disorder that leads to oral mucosa scarring. Aim: In this study, we aimed to study and compare the differences in the buccal cytological smear and the nuclear and cytoplasmic changes seen in the exfoliated cells of patients with OSMF and healthy controls, during preliminary screening. Materials and Methods: Patients with clinical OSMF (reduced mouth opening, blanched buccal mucosa, burning sensation in the buccal mucosa, palpable vertical bands) were included in the study. The cytological smears were stained with Papanicolaou stain and studied under light microscopy. The nuclear and cytoplasmic changes were recorded. The cytologic features that were assessed included the presence of keratinized squames, intermediate cells, inflammatory cells, cytoplasmic creasing, peri nuclear halo, and abnormal chromatin. In addition, the presence and absence of candidal organisms in the smear were assessed. Results: The cytological features which were seen to predominate in the OSMF patients were cells of intermediate and keratinized buccal epithelial cells, cytoplasmic creasing, and presence of inflammatory cells. Conclusion: We conclude that the results of our study indicate that there are cytological changes associated with OSMF that can be assessed with exfoliative cytology. The cellular pattern alterations recorded in our study adds on to the current knowledge pool and literature on OSMF. Further studies are necessary to validate the significance of such cellular alterations.

First total synthesis of chromanone A, preparation of related compounds and evaluation of their antifungal activity against Candida albicans, a biofilm forming agent.

A straightforward and convenient approach for the first total syntheses of chromanone A and a related 7-OMe substituted natural product is reported. These unique C-3 substituted 2-hydroxymethyl chromones were recently isolated as fungal metabolites. Chromanone A was synthesized in 25.3% overall yield from the readily available pyrocatechol, whereas the second natural product was prepared in 39.7% global yield. A small library of chromones, including both natural products and some of their synthetic heterocyclic precursors, was evaluated against Candida albicans ATCC 10231, a biofilm forming agent. It was found that 8-methoxy-3-methyl-4-oxo-4H-chromene-2-carbaldehyde, a partially oxidized form of chromanone A, exhibited a minimum inhibitory concentration of 7.8 μg mL−1 and significantly inhibited the yeast's virulence factors, including the adherence to buccal epithelial cells and the secretion of phospholipases, as well as the formation of germ tubes and the generation of the hyphal pseudomycelium. In addition, despite the heterocycle exhibiting non-significant inhibition of the formation of the Candida biofilm, it completely inhibited the growth of C. albicans in preformed biofilms at 62.5 μg mL−1.

Відповідь ізольованих клітин букального епітелію людини на спільну дію протипухлинного антибіотика доксорубіцину і магнітного поля

The combined effect of doxorubicin and magnetic field on the viability of isolated buccal epithelial cells two donors was investigated. Doxorubicin is an effective antitumor antibiotic, but this drug has a large amount of side effects. This significantly reduces the number of cases when you can change the drug. To reduce the toxic effect or to increase the effectiveness of anticancer therapy, the introduction of additional substances is used. For a long time, magnetic and electromagnetic fields have been used as a therapeutic factor by three therapies for cancer alone, or as an adjunct. Doxorubicin at a concentration of 2 μg/ml for 2 hours has been shown to increase of cell membrane permeability and heterochromatin granule quantity in cell nuclei. Exposure of cells to 25 mT magnetic field for 30 and 60 minutes results in decrease in cell viability and increase in cell membrane permeability and the number of heterochromatin granules in nuclei. Combined exposure to static magnetic field and doxorubicin results in decrease of the number of heterochromatin granules in the nuclei of cells from one of the donors, as compared to the variant of exposure only to doxorubicin, which indicates the protective effect of the magnetic field. A possible mechanism of the protective action of the magnetic field in our experiments is that the stress under the influence of the magnetic field activates the protective mechanisms of the cell (including the transition of chromatin to a heterochromatinized state). That way, been pre-activated due to the action of the magnetic field, the defense mechanisms of the cell reduce the toxic effects caused by doxorubicin.

Цитобіофізична методика визначення індивідуальної чутливості до НВЧ-терапії

Розроблено й досліджено цитобіофізичну методику визначення індивідуальної чутливості до надвисокочастотної терапії в неврології. Проведено дослідження 55 пацієнтів із діагнозом дисциркуляторної енцефалопатії I і II ступенів. Показано неоднозначну реакцію клітин букального епітелію на надвисокочастотне випромінювання. Застосування цитобіофізичної методики дозволяє на клітинному рівні визначати індивідуальну чутливість хворого до надвисокочастотної терапії на всіх етапах лікування.

Применение биоконструкции с культивированными клетками буккального эпителия в лечении пациентов с повреждениями роговицы

Применение биоконструкции с культивированными клетками буккального эпителия в лечении пациентов с повреждениями роговицы

Age and AgNor- A Morphometric study

To investigate and correlate total AgNOR area/Total nucleus area (TAA/TNA ) values in buccal epithelial cells of healthy individuals in different age groups. Material and Methodology:- In present study 50 healthy individuals are included with age ranging 10 to 60 years. These are divided into 5 groups.Group1-10-20year, Group2-20- 30year, Group3-30-40year, Group4-40-50year, Group5-50-60year. Oral epithelial cells collected with the help of cyto brush from buccal mucosa. Smears were prepared on clean glass slides and fixed with 95% alcohol. Fixed slides Smears were stained with AgNOR stain. Results: Statistically significant correlations were found between mean TAA/TNA values and age p < 0.001 for linear and p < 0.0001 for polynominal regression, and between AgNOR number and age, p < 0.001 for linear. Conclusion: There is a significant correlation between age and AgNOR amount (ribosome biosynthesis rate) in buccal epithelial cells of healthy individuals. AgNORs in buccal epithelial cells may be used for detection of age.

Micronucleus Evaluation in Exfoliated Human Buccal Epithelium Cells among E-Waste Workers in Payatas, the Philippines.

Background.The improper recycling of electronic waste (e-waste) by informal recyclers often leads to contamination of the environment. E-waste contains organic and inorganic compounds along with heavy metals and trace elements. These pollutants can have a negative effect on humans. Biomonitoring can provide information on the sources, amount, geographical distribution, and adverse health effects of contaminants.Objectives.The present study aimed to assess risks to the health of informal e-waste recyclers in Payatas, the Philippines due to their exposure to e-waste toxicity by examining the presence of micronuclei in buccal epithelium cells.Methodology.Frequencies of binucleated cells (BNc) and abnormal cells were obtained from the buccal epithelium of the study population composed of e-waste exposed recyclers (n=40) and a control group (n=52). Descriptive statistics and regression analysis were employed for the data analysis.Results.Participants' gender, occupation, smoking status, alcohol consumption, and the number of karyolitic cells of both groups were significantly associated. Only occupation in e-waste recycling and length of e-waste exposure were significantly associated in terms of the number of abnormal cells and micronuclei. Similar trends were found in the linear regression analysis drawn from participants' length of e-waste exposure with a significance of R2= 7346, indicating that as the length of e-waste exposure increased, the number of micronuclei found in the participants' buccal epithelium cells increased as well.Conclusions.Longer exposure to e-waste materials may induce genotoxic damage in human cells which is a serious concern, leading to adverse effects to human health.Competing Interests.The authors declare no competing financial interests.

Assessment of Karyorrhexis Incidence in Exfoliated Buccal Mucosa Epithelial Cells among Fuel Station Employees in Sleman, Special Region of Yogyakarta, Indonesia

Assessment of Karyorrhexis Incidence in Exfoliated Buccal Mucosa Epithelial Cells among Fuel Station Employees in Sleman, Special Region of Yogyakarta, Indonesia

Antagonism of Type I Interferon by Severe Acute Respiratory Syndrome Coronavirus 2.

The coronavirus disease 2019 (COVID-19) pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), warranting urgent study of the molecular mechanisms of SARS-CoV-2 infection and host immune response. Type I interferon (IFN-I) is a key component of host innate immune system responsible for eliminating the virus at the early stage of infection. In contrast, SARS-CoV-2 has evolved multiple strategies to evade innate immune response to facilitate viral replication, transmission, and pathogenesis. This review summarizes the recent progresses on SARS-CoV-2 proteins that antagonize host IFN-I production and/or signaling. These progresses have provided knowledge for new vaccine and antiviral development to prevent and control COVID-19.