ABSTRACT
The extraction and burning of coal release genotoxic pollutants, and understanding the relationship between genetic damage and the spatial distribution of residences in coal-using regions is crucial. The study aimed to conduct a spatial analysis of genotoxic damage through the of micronuclei (MNs) number and their proximity to coal mining/burning in the largest coal exploration region in Brazil. In this study, the detection of genotoxic damage was performed using the MN assay in oral cells of residents exposed to coal mining activities. Spatial analysis was conducted using QGIS 3.28.10 based on information obtained from a questionnaire administered to the population. Multiple linear regression analysis was carried out to assess the influence of the distance from residential areas to polluting sources on the number of MNs found. Additionally, Spearman's correlation was performed to identify the strength and direction of the association between the frequency of MNs and each of the polluting sources. A total of 147 MNs were quantified among all participants in the coal mining region. Notably, residents living within 2â¯km and 10â¯km of pollution sources exhibited the highest prevalence of MNs. The analysis demonstrated a significant correlation between closer proximity to pollution sources and increased MN frequency, underscoring the spatial relationship between these sources and genotoxic damage. Environmental pollutants from anthropogenic sources present a major health risk, potentially leading to irreversible damage. The spatial analysis in this study highlights the importance of targeted public policies. These policies should aim for a sustainable balance between economic development and public health, promoting effective measures to mitigate environmental impacts and protect community health.
Subject(s)
Coal Mining , Micronuclei, Chromosome-Defective , Micronucleus Tests , Mouth Mucosa , Brazil , Humans , Mouth Mucosa/cytology , Micronuclei, Chromosome-Defective/statistics & numerical data , Micronuclei, Chromosome-Defective/chemically induced , Adult , Male , Environmental Exposure/adverse effects , Female , Middle Aged , DNA Damage , Spatial Analysis , Young AdultABSTRACT
The aim of this study was to assess the surface and tissue quality of keratinized mucosa grafts (KMG) obtained using the conventional scalpel and mucotome techniques. This was an experimental in vitro/ex vivo study involving six porcine hemi-mandibles. Specimens were harvested using both the mucotome and conventional scalpel techniques, with randomization determining the choice of technique for tissue removal. The specimens were prepared following predefined laboratory protocols and subsequently subjected to optical microscopy for evaluating epithelial and connective tissue and scanning electron microscopy for topographical and 3D profilometry analysis. Tissues harvested using the mucotome exhibited a linear base and uniform thickness, along with the presence of submucosa and fibrous connective tissue, all of which are ideal for graft success. Differences in the surface characteristics of specimens obtained through the two techniques were observed during a comparative analysis of images obtained through both microscopy types. KMG obtained using the mucotome technique displayed greater uniformity and reduced undesirable cell presence compared to the scalpel technique, thereby enhancing the likelihood of success in soft tissue graft surgical procedures. This study provides valuable insights to oral healthcare professionals and may contribute to future research aimed at achieving more successful surgeries, shorter postoperative recovery times, reduced discomfort, and an overall more positive patient experience.
Subject(s)
Mandible , Mouth Mucosa , Animals , Swine , Mouth Mucosa/transplantation , Mouth Mucosa/cytology , Mandible/surgery , Keratins/metabolism , Microscopy, Electron, Scanning , Tissue and Organ Harvesting/methodsABSTRACT
Insufficient osseointegration of titanium-based implants is a factor conditioning their long-term success. Therefore, different surface modifications, such as multifunctional oxide coatings, calcium phosphates, and the addition of molecules such as peptides, have been developed to improve the bioactivity of titanium-based biomaterials. In this work, we investigate the behavior of human oral mucosal stem cells (hOMSCs) cultured on amorphous titanium oxide (aTiO2), surfaces designed to simulate titanium (Ti) surfaces, biofunctionalized with a novel sequence derived from cementum attachment protein (CAP-p15), exploring its impact on guiding hOMSCs towards an osteogenic phenotype. We carried out cell attachment and viability assays. Next, hOMSCs differentiation was assessed by red alizarin stain, ALP activity, and western blot analysis by evaluating the expression of RUNX2, BSP, BMP2, and OCN at the protein level. Our results showed that functionalized surfaces with CAP-p15 (1 µg ml-1) displayed a synergistic effect increasing cell proliferation and cell attachment, ALP activity, and expression of osteogenic-related markers. These data demonstrate that CAP-p15 and its interaction with aTiO2surfaces promote osteoblastic differentiation and enhanced mineralization of hOMSCs when compared to pristine samples. Therefore, CAP-p15 shows the potential to be used as a therapeutical molecule capable of inducing mineralized tissue regeneration onto titanium-based implants.
Subject(s)
Cell Adhesion , Cell Differentiation , Cell Proliferation , Mouth Mucosa , Osteogenesis , Stem Cells , Titanium , Titanium/chemistry , Humans , Osteogenesis/drug effects , Mouth Mucosa/cytology , Mouth Mucosa/metabolism , Stem Cells/cytology , Stem Cells/metabolism , Surface Properties , Cells, Cultured , Osteoblasts/cytology , Osteoblasts/metabolism , Coated Materials, Biocompatible/chemistry , Coated Materials, Biocompatible/pharmacology , Cell Survival , Osseointegration/drug effects , Biocompatible Materials/chemistryABSTRACT
OBJECTIVE: To analyze the effect of hookah and cigarettes on the oral mucosa of smokers through the use of exfoliative cytology. STUDY DESIGN: Smear samples were collected by exfoliative cytology from the tongue of 33 hookah smokers, 22 cigarette smokers, and 30 non-smokers. The selected analyses include micronuclei (MN), metanuclear anomalies, epithelial maturation, and cytomorphology (nuclear area [NA], cytoplasmic area [CA], and NA/CA ratio). RESULTS: The largest differences observed for MN and metanuclear anomalies were between cigarette smokers and the control group (notably 1 MN P = .04; total cells with MN P = .039; total MN P = .042; karyorrhexis and binucleation, P = .0001). The hookah group, compared with the control group, showed the greatest differences for karyolysis (P = .0023), binucleation (P = .0003), and broken egg (P = .008). Significant differences were found between the smokers and the control groups regarding changes in the superficial cell without nucleus, perinuclear halo, vacuolization, color change, mucus, and keratohyalin granules. There was a significant increase in the NA and NA/CA ratio in the smoker groups. CONCLUSION: This study showed that a combined analysis of exfoliative cytology associated with other diagnostic methods is a useful tool for studying oral carcinogenesis. Hookah and cigarettes showed similar effects in terms of displaying substantial cytogenetic and cytotoxic damage.
Subject(s)
Micronucleus Tests , Mouth Mucosa , Humans , Mouth Mucosa/pathology , Mouth Mucosa/cytology , Male , Female , Adult , Middle Aged , Micronuclei, Chromosome-Defective , Smoking/adverse effects , Cytodiagnosis/methods , Cigarette Smoking/adverse effects , Case-Control StudiesABSTRACT
BACKGROUND: Diagnosing imprinting defects in neonates and young children presents challenges, often necessitating molecular analysis for a conclusive diagnosis. The isolation of genetic material from oral swabs becomes crucial, especially in settings where blood sample collection is impractical or for vulnerable populations like newborns, who possess limited blood volumes and are often too fragile for invasive procedures. Oral swab samples emerge as an excellent source of DNA, effectively overcoming obstacles associated with rare diseases. METHODS: In our study, we specifically addressed the determination of the quality and quantity of DNA extracted from oral swab samples using NaCl procedures. RESULTS: We compared these results with extractions performed using a commercial kit. Subsequently, the obtained material underwent MS-HRM analysis for loci associated with imprinting diseases such as Prader-Willi and Angelman syndromes. CONCLUSIONS: Our study emphasizes the significance of oral swab samples as a reliable source for obtaining DNA for MS-HRM analysis. NaCl extraction stands out as a practical and cost-effective method for genetic studies, contributing to a molecular diagnosis that proves particularly beneficial for patients facing delays in characterization, ultimately influencing their treatment.
Subject(s)
Angelman Syndrome , DNA , Genomic Imprinting , Mouth Mucosa , Prader-Willi Syndrome , Humans , Mouth Mucosa/cytology , Mouth Mucosa/pathology , Angelman Syndrome/genetics , Angelman Syndrome/diagnosis , Prader-Willi Syndrome/genetics , Prader-Willi Syndrome/diagnosis , DNA/genetics , DNA/isolation & purification , Sodium Chloride , Infant, Newborn , Male , Imprinting DisordersABSTRACT
This study aimed to evaluate all studies which used the micronucleus assay using oral cells in the attempt to understand whether such technique is efficient in evaluating genotoxicity in gas station attendants. Full manuscripts from 16 studies were carefully selected by the authors. Our results demonstrate that continuous exposure to derivatives of petroleum may lead to genotoxic effects since all studies demonstrated positive findings (16 out of 16) and 11 of them had a strong or moderate final rating. In summary, our results reveal that gas station attendants are occupationally exposed to genotoxic agents and that the micronucleus assay in oral mucosa is indeed an effective method to evaluate genotoxicity in this specific case. Such findings are very important for protecting these professionals who are continuously exposed to chemicals for long periods.
Subject(s)
Micronucleus Tests , Mouth Mucosa , Occupational Exposure , Micronucleus Tests/methods , Humans , Occupational Exposure/adverse effects , Mouth Mucosa/drug effects , Mouth Mucosa/cytology , Carcinogenesis/chemically induced , Carcinogenesis/drug effects , Biomarkers/analysis , Petroleum/toxicity , Risk Assessment/methodsABSTRACT
Human oral mucosa stem cells (hOMSCs) arise from the neural crest, they can self-renew, proliferate, and differentiate to several cell lines and could represent a good source for application in tissue engineering. Because of their anatomical location, hOMSCs are easy to isolate, have multilineage differentiation capacity and express embryonic stem cells markers such as-Sox2, Oct3/4 and Nanog. We have used SHEM (supplemented hormonal epithelial medium) media and cultured hOMSCs over human amniotic membrane and determined the cell's capacity to differentiate to an epithelial-like phenotype and to express corneal specific epithelial markers-CK3, CK12, CK19, Pan-cadherin and E-cadherin. Our results showed that hOMSCs possess the capacity to attach to the amniotic membrane and express CK3, CK19, Pan-Cadherin and E-Cadherin without induction with SHEM media and expressed CK12 or changed the expression pattern of E-Cadherin to a punctual-like feature when treated with SHEM media. The results observed in this study show that hOMSCs possess the potential to differentiate toward epithelial cells. In conclusion, our results revealed that hOMSCs readily express markers for corneal determination and could provide the ophthalmology field with a therapeutic alternative for tissue engineering to achieve corneal replacement when compared with other techniques. Nevertheless, further studies are needed to develop a predictable therapeutic alternative for cornea replacement.
Subject(s)
Cell Differentiation/genetics , Epithelium, Corneal/growth & development , Mesenchymal Stem Cells/cytology , Mouth Mucosa/growth & development , Amnion/growth & development , Cells, Cultured , Cornea/cytology , Cornea/growth & development , Cornea/metabolism , Culture Media/pharmacology , Embryonic Stem Cells/cytology , Embryonic Stem Cells/metabolism , Epithelial Cells/cytology , Epithelial Cells/metabolism , Epithelium, Corneal/cytology , Gene Expression Regulation, Developmental/genetics , Humans , Mouth Mucosa/cytology , Tissue Engineering/trendsABSTRACT
Different body systems (epidermis, respiratory tract, cornea, oral cavity, and gastrointestinal tract) are in continuous direct contact with innocuous and/or potentially harmful external agents, exhibiting dynamic and highly selective interaction throughout the epithelia, which function as both a physical and chemical protective barrier. Resident immune cells in the epithelia are constantly challenged and must distinguish among antigens that must be either tolerated or those to which a response must be mounted for. When such a decision begins to take place in lymphoid foci and/or mucosa-associated lymphoid tissues, the epithelia network of immune surveillance actively dominates both oral and gastrointestinal compartments, which are thought to operate in the same immune continuum. However, anatomical variations clearly differentiate immune processes in both the mouth and gastrointestinal tract that demonstrate a wide array of independent immune responses. From single vs. multiple epithelia cell layers, widespread cell-to-cell junction types, microbial-associated recognition receptors, dendritic cell function as well as related signaling, the objective of this review is to specifically contrast the current knowledge of oral versus gut immune niches in the context of epithelia/lymphoid foci/MALT local immunity and systemic output. Related differences in 1) anatomy 2) cell-to-cell communication 3) antigen capture/processing/presentation 4) signaling in regulatory vs. proinflammatory responses and 5) systemic output consequences and its relations to disease pathogenesis are discussed.
Subject(s)
Allostasis , Homeostasis , Immunity, Mucosal/immunology , Immunologic Surveillance/immunology , Intestinal Mucosa/immunology , Mouth Mucosa/immunology , Adaptive Immunity , Animals , Antigen Presentation , Bacterial Translocation/immunology , Cell Adhesion Molecules/physiology , Cell Communication , Dendritic Cells/immunology , Dysbiosis/immunology , Epithelial Cells/immunology , Humans , Inflammation , Intercellular Junctions/physiology , Intestinal Mucosa/cytology , Microbiota , Mouth Mucosa/cytology , Mucus/physiology , Organ Specificity , Saliva/immunology , Signal TransductionABSTRACT
The Buccal Micronucleus Cytome Assay (BMCyt) has become an important biomonitoring tool for assessing cytogenetic damage in many studied populations. Each laboratory applies protocols that vary according to the method of collecting and preparing samples. Besides, Brazil is a country of great territorial extensions that received immigrants from various parts of the world with different genetic backgrounds. Therefore, the present study aimed to evaluate the inter-laboratory variation in scoring the same set of slides using the more comprehensive scoring criteria, to standardize the BMCyt protocol, to observe the basal alterations in populations of different Brazilian regions and to compare it with other places around the world. Our results showed that a valuable number of laboratories participated, ten laboratories from different regions of the country, for the validation of the BMCyt in human biomonitoring studies, resulting in the 804 healthy individuals. This was possible because we observed: a range of measures needs to be considered, such as the baseline frequency of DNA damage and cell death in non-exposed individuals; age when grouped showed an influence on DNA damage, although when evaluated by group we did not see an influence; association between smoking habit and all endpoints of the BMCyt (except karyolytic cells) was evident; the basal MN frequency, in the majority of groups, follows those around the world; and the BMCyt was confirmed as a good health status biomarker. We emphasize the need for constant discussions on the parameters of cell death due to greater difficulty among the analyzers.
Subject(s)
Biological Assay/standards , Cell Nucleus/genetics , Epithelial Cells/ultrastructure , Laboratories/standards , Micronuclei, Chromosome-Defective , Micronucleus Tests/standards , Mouth Mucosa/cytology , Adolescent , Adult , Biological Assay/methods , Brazil , Cell Death/genetics , Cell Nucleus/ultrastructure , DNA Damage , Female , Humans , Male , Micronuclei, Chromosome-Defective/statistics & numerical data , Micronucleus Tests/methods , Middle Aged , Mouth Mucosa/ultrastructure , Young AdultABSTRACT
BACKGROUND: The herbicide dichlorophenoxyacetic acid (2,4-D) is one of the most widely used crop spraying products in the world. Some pesticides induce the degranulation of mast cells and increase allergic responses. This is the first study to evaluate the damage to the oral mucosa after an experimental simulation of environmental inhalation exposure to the 2,4-D herbicide. The aim of this study was evaluate the possible oral damage caused by acute inhalation exposure to the herbicide 2,4-D. RESULTS: There was a difference between the exposure concentrations in relation to tissue congestion intensity (p = 0.002) and mast cell counts (p = 0.002), a difference in the evaluation of the interaction between the exposure concentrations and nebulization time in the dorsum epithelium thickness (p = 0.013), and a significant correlation between the epithelial thickness and the number of nucleoli organizing regions on the dorsum of the tongue (p = 0.048). CONCLUSIONS: Even after acute exposure, the herbicide 2,4-D had the potential to damage the oral epithelium, especially at higher doses.
Subject(s)
2,4-Dichlorophenoxyacetic Acid/toxicity , Herbicides/toxicity , Inhalation Exposure/adverse effects , Mouth Mucosa/pathology , Animals , Epithelium/pathology , Male , Mast Cells , Mice , Mouth Mucosa/cytology , Nucleolus Organizer Region , Tongue/cytology , Tongue/pathologyABSTRACT
The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins ß1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins ß1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.
Subject(s)
Cell Culture Techniques/methods , Epithelial Cells/cytology , Keratinocytes/cytology , Mouth Mucosa/cytology , Phenotype , Stem Cells/cytology , Antigens, CD/analysis , Biomarkers/analysis , Cell Separation/methods , Flow Cytometry/methods , Humans , Nerve Tissue Proteins/analysis , Receptors, Nerve Growth Factor/analysis , Receptors, Transferrin/analysis , Reproducibility of ResultsABSTRACT
Background: Micronucleus is a microscopically visible cyto-plasmic chromatin mass in the extranuclear vicinity, originating from aberrant mitosis, which consists of eccentric chromosomes that have failed to reach spindle poles during mitosis. The present study was designed to evaluate and compare cytogenetic changes in the buccal mucosa of smokers and non-smokers based on the occurrence of micronuclei. The study aimed to determine the correlation between the micronuclei count and the frequency and duration of smoking habit. Materials and Methods: Two groups (smokers and non-smokers) of 34 individuals each were examined. Cytological buccal smears were taken from participants using a moistened wooden spatula and stained with standard Papanicolaou stain. Presence of micronuclei was assessed at 40X magnification using a light microscope and a count per 500 cells was determined. The results of the study were analyzed statistically using Mann-Whitney U test, Spearman's rank correlation coefficient and Student t-test. Result: Smears from smokers showed a significant increase in the total number of micronuclei per 500 cell count compared to non-smokers. There was a strong positive correlation between the occurrence of micronuclei and the frequency and duration of smoking. A age-related increase in older age groups was also observed. Conclusion: The study reveals a strong positive correlation between the occurrence of micronuclei and the frequency and duration of smoking. This observation is vital in the utilization of the micronuclei detection in smears as a prognostic, educational and interventional tool in the management of patients with smoking habits.
Antecedentes: El micronúcleo es una masa de cromatina citoplasmática microscópicamente visible en el área extranuclear, que se origina a partir de la mitosis aberrante, y que consiste en cromosomas excéntricos que no han podido alcanzar los polos del huso durante la mitosis. El presente estudio fue diseñado para evaluar y comparar los cambios citogenéticos en la mucosa bucal de fumadores y no fumadores en función de la aparición de micronúcleos. El estudio tuvo como objetivo determinar la correlación entre el recuento de micronúcleos y la frecuencia y duración del hábito de fumar. Materiales and Métodos: Se examinaron dos grupos (fumadores y no fumadores) de 34 individuos cada uno. Se tomaron frotis bucales citológicos de todos los participantes con una espátula de madera humedecida y se tiñeron con la tinción estándar de Papanicolaou. La presencia de micronúcleos se evaluó al microscopio óptico con un aumento de 40X y se determinó un recuento por 500 células. Los resultados del estudio se analizaron estadísticamente utilizando la prueba U de Mann-Whitney, el coeficiente de correlación de rango de Spearman y la prueba t de Student. Resultados: Los frotis de fumadores mostraron un aumento significativo en el número total de micronúcleos por 500 células en comparación con los no fumadores. Hubo una fuerte correlación positiva entre la aparición de micronúcleos y la frecuencia y duración del tabaquismo. También se observó un aumento relacionado con la edad en los grupos de mayor edad. Conclusión: el estudio revela una fuerte correlación positiva entre la aparición de micronúcleos y la frecuencia y duración del tabaquismo. Esta observación es vital en la utilización de la detección de micronúcleos en frotis como una herramienta pronostica, educativa e intervencionista en el manejo de pacientes con hábitos de fumar.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Young Adult , Micronucleus Tests , Micronuclei, Chromosome-Defective , Tobacco Use/adverse effects , Tobacco Smoking/adverse effects , Mouth Mucosa/cytology , In Vitro Techniques , Chromosome Aberrations , Non-Smokers , IndiaABSTRACT
In the present study, the morphological development of the Brycon amazonicus digestive tract is described to provide basic knowledge for nutritional studies and, therefore, increase the survival of this species during larviculture. Samples were collected from hatching up to 25 days of age, measured, processed and observed under a stereomicroscope and light microscopy. Newly hatched larvae presented their digestive tract as a straight tube, dorsal to the yolk sac, lined with a single layer of undifferentiated cells. At 24 h post-hatching (hPH), the buccopharyngeal cavity was open, but the posterior region of the digestive tube remained closed. At 25 hPH, the digestive tube was completely open and could be divided into buccopharyngeal cavity, oesophagus and intestine. At 35 hPH, the intestine presented a dilatation in the proximal region, which had the function of storing food. Differentiation of the stomach started at 83 hPH, and mucous cells were observed in the epithelium. These cells are important in the production of mucus, whose function is to protect the organ against acidity, although the gastric glands began developing only from 171 hPH, when three stomach regions were observed: cardiac, fundic and pyloric. The gastric glands were observed in the cardiac region, indicating that this organ already had digestive functionality. From 243 hPH, the absorption and assimilation of nutrients were already possible but, only from 412 hPH, the digestive tract was completely developed and functional.
Subject(s)
Characiformes/growth & development , Gastrointestinal Tract/growth & development , Animals , Branchial Region/cytology , Branchial Region/embryology , Branchial Region/growth & development , Characiformes/anatomy & histology , Characiformes/embryology , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/embryology , Embryonic Development/physiology , Gastric Mucosa/cytology , Gastric Mucosa/embryology , Gastric Mucosa/growth & development , Gastrointestinal Tract/cytology , Gastrointestinal Tract/embryology , Larva/cytology , Larva/growth & development , Mouth Mucosa/cytology , Mouth Mucosa/embryology , Mouth Mucosa/growth & development , Time FactorsABSTRACT
Human cementum protein 1 (CEMP1) is known to induce cementoblast and osteoblast differentiation and alkaline phosphatase (ALP) activity in human periodontal ligament-derived cells in vitro and promotes bone regeneration in vivo. CEMP1's secondary structure analysis shows that it has a random-coiled structure and is considered an Intrinsic Disordered Protein (IDP). CEMP1's short peptide sequences mimic the biological capabilities of CEMP1. However, the role and mechanisms of CEMP1's C-terminal-derived synthetic peptide (CEMP1-p4) in the canonical Wnt/ß-catenin signaling pathway are yet to be described. Here we report that CEMP1-p4 promotes proliferation and differentiation of Human Oral Mucosa Stem Cells (HOMSCs) by activating the Wnt/ß-catenin pathway. CEMP1-p4 stimulation upregulated the expression of ß-catenin and glycogen synthase kinase 3 beta (GSK-3B) and activated the transcription factors TCF1/7 and Lymphoid Enhancer binding Factor 1 (LEF1) at the mRNA and protein levels. We found translocation of ß-catenin to the nucleus in CEMP1-p4-treated cultures. The peptide also penetrates the cell membrane and aggregates around the cell nucleus. Analysis of CEMP1-p4 secondary structure revealed that it has a random-coiled structure. Its biological activities included the induction to nucleate hydroxyapatite crystals. In CEMP1-p4-treated HOMSCs, ALP activity and calcium deposits increased. Expression of Osterix (OSX), Runt-related transcription factor 2 (RUNX2), Integrin binding sialoproptein (IBSP) and osteocalcin (OCN) were upregulated. Altogether, these data show that CEMP1-p4 plays a direct role in the differentiation of HOMSCs to a "mineralizing-like" phenotype by activating the ß-catenin signaling cascade.
Subject(s)
Mouth Mucosa/growth & development , Osteogenesis/genetics , Periodontal Ligament/growth & development , Proteins/chemistry , Stem Cells/cytology , Bone Regeneration/genetics , Cell Differentiation/drug effects , Cell Proliferation/genetics , Core Binding Factor Alpha 1 Subunit/genetics , Dental Cementum/metabolism , Durapatite/metabolism , Gene Expression Regulation, Developmental/genetics , Glycogen Synthase Kinase 3 beta/genetics , Humans , Integrin-Binding Sialoprotein/genetics , Mouth Mucosa/cytology , Mouth Mucosa/metabolism , Osteoblasts/metabolism , Osteocalcin/genetics , Peptides/chemistry , Peptides/genetics , Periodontal Ligament/cytology , Periodontal Ligament/metabolism , Protein Structure, Secondary , Proteins/genetics , Proteins/ultrastructure , Sp7 Transcription Factor/genetics , Stem Cells/metabolism , Wnt Signaling Pathway/geneticsABSTRACT
Abstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.
Subject(s)
Humans , Phenotype , Stem Cells/cytology , Keratinocytes/cytology , Cell Culture Techniques/methods , Epithelial Cells/cytology , Mouth Mucosa/cytology , Receptors, Transferrin/analysis , Biomarkers/analysis , Antigens, CD/analysis , Cell Separation/methods , Reproducibility of Results , Receptors, Nerve Growth Factor/analysis , Flow Cytometry/methods , Nerve Tissue Proteins/analysisABSTRACT
The presence of Candida albicans in the biofilm underlying the dental prosthesis is related to denture stomatitis (DS), an inflammatory reaction of the oral mucosa. The oral epithelium, a component of the innate immune response, has the ability to react to fungal invasion. In this study, we evaluated the in vitro effect of viable C. albicans on the apoptosis, nitric oxide (NO) production, and ß-defensin 2 (hBD-2) expression and production of human palate epithelial cells (HPECs). We further determined whether or not these effects were correlated with fungal invasion of epithelial cells. Interaction between HPEC primary culture and C. albicans was obtained through either direct or indirect cell-cell contact with a supernatant from a hyphal fungus. We found that the hyphae supernatants were sufficient to induce slight HPEC apoptosis, which occurred prior to the activation of the specific mechanisms of epithelial defense. The epithelial defense responses were found to occur via NO and antimicrobial peptide hBD-2 production only during direct contact between C. albicans and HPECs and coincided with the fungus's intraepithelial invasion. However, although the hBD-2 levels remained constant in the HPEC supernatants over time, the NO release and hBD-2 gene expression were reduced at a later time (10 h), indicating that the epithelial defense capacity against the fungal invasion was not maintained in later phases. This aspect of the immune response was associated with increased epithelial invasion and apoptosis maintenance.
Subject(s)
Fibroblasts , Keratinocytes , Mouth Mucosa , Nitric Oxide/metabolism , Palate , beta-Defensins/metabolism , Biofilms , Candida albicans/physiology , Candidiasis/immunology , Candidiasis/microbiology , Cell Line , Fibroblasts/cytology , Fibroblasts/metabolism , Host-Pathogen Interactions , Humans , Immunity, Innate , Keratinocytes/cytology , Keratinocytes/metabolism , Mouth Mucosa/cytology , Mouth Mucosa/metabolism , Palate/cytology , Palate/metabolismABSTRACT
Soybean farmers are exposed to various types of pesticides that contain in their formulations a combination of chemicals with genotoxic and mutagenic potential. Therefore, the objective of this paper was to evaluate the genetic damages caused by this pesticide exposure to soybean producers in the state of Mato Grosso (Brazil), regarding biochemical, genetic polymorphic and in silico analyses. A total of 148 individuals were evaluated, 76 of which were occupationally exposed and 72 were not exposed at all. The buccal micronucleus cytome assay (BMCyt) detected in the exposed group an increase on DNA damage and cell death. No inhibition of butyrylcholinesterase (BchE) was observed within the exposed group. The detection of inorganic elements was made through the particle-induced X-ray emission technique (PIXE), which revealed higher concentrations of Bromine (Br), Rubidium (Rb) and Lead (Pb) in rural workers. A molecular model using in silico analysis suggests how metal ions can cause both DNA damage and apoptosis in the exposed cells. Analysis of the compared effect of X-ray Repair Cross-complement Protein 1 (XRCC1) and Paraoxonase 1 (PON1) genotypes in the groups demonstrated an increase of binucleated cells (exposed group) and nuclear bud (non-exposed group) in individuals with the XRCC1 Trip/- and PON1 Arg/- genes. There was no significant difference in the telomere (TL) mean value in the exposed group in contrast to the non-exposed group. Our results showed that soybean producers showed genotoxic effect and cell death, which may have been induced by exposure to complex mixtures of agrochemicals and fertilizers. In addition, XRCC1 Arg/Arg could, in some respects, provide protection to individuals.
Subject(s)
Complex Mixtures/toxicity , DNA Damage , Fertilizers/toxicity , Micronuclei, Chromosome-Defective/chemically induced , Occupational Exposure/adverse effects , Pesticides/toxicity , Polymorphism, Genetic , Adult , Apoptosis/drug effects , Aryldialkylphosphatase/drug effects , Brazil , Computer Simulation , Epithelial Cells/drug effects , Farmers , Genotype , Humans , Male , Middle Aged , Mouth Mucosa/cytology , Occupational Exposure/analysis , X-ray Repair Cross Complementing Protein 1/geneticsABSTRACT
BACKGROUND/AIM: Studies have demonstrated the biological consequences of environmental contamination caused by human pesticide exposure following banana production. The aim of this study was to evaluate genomic instability and cytotoxicity in buccal mucosal cells of workers in banana farming. MATERIALS AND METHODS: For this purpose, a total of 21 male workers in banana farming in the Ribeira Valley were included in the experimental group. A total of 20 individuals, not occupationally exposed to pesticides, were included in the control group. RESULTS: The frequency of micronuclei was significantly increased (p<0.05) in buccal mucosa cells from workers of banana farming when compared to the control group. Furthermore, a high frequency of karyolysis was detected in buccal mucosaI cells in these individuals. No significant differences were found in pyknosis or karryorhexis when compared to controls. CONCLUSION: Taken together, our results indicate that workers in banana farming represent a group in high risk for carcinogenesis since chromosomal damage and cellular death are increased in these individuals.
Subject(s)
Micronuclei, Chromosome-Defective/chemically induced , Mouth Mucosa/cytology , Musa , Occupational Exposure/adverse effects , Pesticides/toxicity , Adult , Brazil , Genomic Instability , Humans , Male , Micronucleus TestsABSTRACT
Periodontitis is a bacterial infection characterized by the presence of a dense inflammatory infiltrate, which may result in increased DNA damage and other nuclear/cellular abnormalities. Therefore, it is important to evaluate the periodontal diseases influence on DNA damage and other nuclear/cellular abnomalies formation as cancer risk markers. Thus, the aim of this study was to evaluate the periodontal diseases effect, according to its severity, on the occurrence of DNA damage and other nuclear/cellular abnormalities. This is a cross-sectional study with 77 subjects from the dentistry clinic of the University of Santa Cruz do Sul, Brazil, divided in control group (26 subjects), moderate periodontal disease group (26 subjects) and severe periodontal disease group (25 subjects). All subjects answered self-referenced questionnaires, underwent periodontal clinical examinations and allowed the collection of oral mucosa cells for the BMCyt. In relation to DNA damage biomarkers (micronuclei (MN) and/or nuclear buds (NBUD)), our results indicated no increase in MN frequencies (p > 0.05), however it indicated significant difference in NBUD frequencies between groups (p < 0.024). This result suggests that the periodontal disease status may influence DNA damage. Regarding the other nuclear/cellular abnormalities, was observed a significant difference in the binucleated (BN) frequencies between groups (p < 0.05). Moreover, the periodontitis severity was associated to an increase in the combined (summed) frequency of cells with different levels of DNA damage (MN and/or NBUD), cytokinetic defects (BN cells) and/or cell death (karyorrhexis, pyknotic and karyolytic cells) (r = 0.235; p = 0.040). Periodontal disease depending on its severity, induces nuclear anomalies in buccal cells.
Subject(s)
DNA Damage/genetics , Genomic Instability/genetics , Mouth Mucosa/cytology , Periodontitis/genetics , Adult , Aged , Brazil , Cross-Sectional Studies , Female , Humans , Male , Micronuclei, Chromosome-Defective , Micronucleus Tests , Middle Aged , Periodontitis/microbiology , Periodontitis/pathology , Reactive Oxygen Species/metabolism , Surveys and QuestionnairesABSTRACT
Contamination with mining wastes affects the environmental health and public, especially the human populations that live in these environments. The aim of this study was to evaluate the genotoxicity and levels of mercury (Hg) and arsenic (As) in blood samples from human populations exposed to mining activities in the upper basin of the San Jorge River. A total of 100 individuals participated in the study, 50 as an exposed group (Bocas de Ure = 15 individuals, Mina el Alacrán = 19 individuals, Torno Rojo = 16 individuals) and 50 individuals participated as the control group. Hg and As contents in blood samples were analyzed with atomic absorption spectrophotometry. A comet assay in peripheral blood lymphocytes and a micronucleus (MN) cytome assay (BMCyt) in exfoliated buccal cells were used to assess the effects of exposure to heavy metals on human communities located in mining areas. Higher concentrations of Hg and As were observed in human populations located in mining areas. The comet assay and BMCyt data revealed DNA damage and cell death in human communities located in mining areas. A positive association between blood arsenic and genetic damage was found. These data confirm the public health risk of the population near mining sites. Our findings suggest that populations that live at sites close to mining activities have high contents of heavy metals and genotoxic effects, representing a risk to human health.