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1.
Braz Oral Res ; 34: e033, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32267290

RESUMO

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.


Assuntos
Técnicas de Cultura de Células/métodos , Células Epiteliais/citologia , Queratinócitos/citologia , Mucosa Bucal/citologia , Fenótipo , Células-Tronco/citologia , Antígenos CD/análise , Biomarcadores/análise , Separação Celular/métodos , Citometria de Fluxo/métodos , Humanos , Proteínas do Tecido Nervoso/análise , Receptores de Fator de Crescimento Neural/análise , Receptores da Transferrina/análise , Reprodutibilidade dos Testes
2.
Mutat Res ; 852: 503159, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32265039

RESUMO

Dental technicians may be chronically exposed to methyl methacrylate (MMA), used in the production of dental prostheses. We have studied whether occupational exposure to MMA affects genotoxicity biomarkers such as 8-OHdG formation, comet assay, and buccal micronucleus frequency. MMA exposure was assessed via ambient air analysis. Although no significant differences between exposed and non-exposed individuals were seen with respect to blood genotoxicity measurements, we found a higher level of buccal-cell anomalies in the exposed group.


Assuntos
Cimentos para Ossos/toxicidade , Técnicos em Prótese Dentária , Metilmetacrilato/toxicidade , Micronúcleos com Defeito Cromossômico/efeitos dos fármacos , Mucosa Bucal/efeitos dos fármacos , 8-Hidroxi-2'-Desoxiguanosina/sangue , Adulto , Estudos de Casos e Controles , Ensaio Cometa , Dano ao DNA , Feminino , Humanos , Masculino , Testes para Micronúcleos , Mucosa Bucal/citologia , Exposição Ocupacional/análise
3.
Int J Occup Environ Med ; 11(1): 33-40, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31905193

RESUMO

BACKGROUND: Workers in cement warehouses of Kerala are enduring long-standing exposure to cement dust, which is considered genotoxic. OBJECTIVE: To evaluate the extent of genotoxicity and cytotoxicity caused due to exposure of cement dust among those working in cement warehouses. METHODS: The study included 82 cement warehouse workers and 82 age-matched individuals with no exposure to cement dust. Exfoliated buccal micronucleus cytome assay (BMCyt) was performed to analyze the genotoxic and cytotoxic effects caused by inhalation of cement dust. RESULTS: The frequency of various genotoxic and cytotoxic end markers (micronucleated cells [2-fold increase, p<0.001], nuclear buds [4-fold increase, p<0.001], binucleated cells [4-fold increase, p<0.001], karyorrhectic cells [2-fold increase, p<0.001], pyknotic cells [3-fold increase, p<0.001], and karyolytic cells [2-fold increase, p<0.001]) were higher in the exposed workers compared with unexposed group. Increase of these parameters represented an increased level of chromosomal damage, nuclear disintegration and increased cell death among exposed group compared with unexposed group. CONCLUSION: Continuous exposure to cement dust results in increased frequency of nuclear aberrations and cellular apoptosis. This may lead to defects in genome maintenance, accelerated ageing, increased chance of oral cancer and neurodegenerative disorders in those occupationally exposed to cement dust.


Assuntos
Dano ao DNA/efeitos dos fármacos , Poeira , Instabilidade Genômica/genética , Mucosa Bucal/citologia , Exposição Ocupacional/efeitos adversos , Adulto , Apoptose/efeitos dos fármacos , Aberrações Cromossômicas/induzido quimicamente , Humanos , Masculino , Testes para Micronúcleos/métodos , Pessoa de Meia-Idade
4.
Anticancer Res ; 39(12): 6673-6684, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31810932

RESUMO

BACKGROUND/AIM: The aim of the study was to evaluate the antitumor potential and combination effects of chemotherapeutic drugs. MATERIALS AND METHODS: The cytotoxicity of 20 drip-type classical and molecular-targeted anticancer drugs was examined against 4 human oral squamous cell carcinoma cell lines and 5 human oral normal mesenchymal and epithelial cells. Cell cycle progression was monitored by a cell sorter. Combination effect was evaluated by combination index. RESULTS: Most of the classical anticancer drugs showed much higher antitumor activity than molecular-targeted drugs, except bortezomib. Among 12 classical anticancer drugs, taxanes and gemsitabine showed the highest tumor-specificity (TS) and potency-selectivity expression (PSE) values, whereas platinum analogs showed the least TS value. Combination of two classical or a classical and a molecular-targeted drug showed mostly additive or antagonistic effect. 5-FU and cisplatin did not produce a subG1 population, but induced G2/M or G1/S arrest, regardless of the addition of cetuximab. Cetuximab, nibolumab and bortezomib showed potent keratinocyte toxicity. CONCLUSION: The present TS monitoring system may provide useful information for building up the treatment regimens of anticancer drugs.


Assuntos
Antineoplásicos/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Carcinoma de Células Escamosas/tratamento farmacológico , Células Epiteliais/efeitos dos fármacos , Terapia de Alvo Molecular , Neoplasias Bucais/tratamento farmacológico , Bortezomib/farmacologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Cetuximab/farmacologia , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Fibroblastos/efeitos dos fármacos , Fluoruracila/farmacologia , Hormese , Humanos , Técnicas In Vitro , Queratinócitos/efeitos dos fármacos , Mucosa Bucal/citologia , Nivolumabe/farmacologia , Compostos de Platina/farmacologia , Taxoides/farmacologia
5.
J Coll Physicians Surg Pak ; 29(10): 1012-1014, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31564282

RESUMO

The aim of this study was to assess the morphological changes of oral mucosa epithelial cells in contact with fixed orthodontic appliances using exfoliated cytology. This study was conducted on 100 individuals that were divided into two groups (50 each). Buccal cytological smears were collected from the control group and case group at 3 to 6 months after the application of orthodontic appliances. Slides were stained with papanicolaou stain and analysed using a light microscope. In the case group, 43 out of 50 patients (86%) exhibited morphological changes which included variable degrees of atrophy in epithelial cells; nuclear fragments and inflammatory cells infiltrate. Epithelial cells showed some morphological changes in patients with orthodontic appliances, which represent an adaptive response.


Assuntos
Células Epiteliais/citologia , Mucosa Bucal/citologia , Aparelhos Ortodônticos Fixos , Feminino , Humanos , Masculino , Adulto Jovem
6.
Nat Commun ; 10(1): 4496, 2019 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-31582750

RESUMO

Solitary chemosensory cells (SCCs) are epithelial sentinels that utilize bitter Tas2r receptors and coupled taste transduction elements to detect pathogenic bacterial metabolites, triggering host defenses to control the infection. Here we report that SCCs are present in mouse gingival junctional epithelium, where they express several Tas2rs and the taste signaling components α-gustducin (Gnat3), TrpM5, and Plcß2. Gnat3-/- mice have altered commensal oral microbiota and accelerated naturally occurring alveolar bone loss. In ligature-induced periodontitis, knockout of taste signaling molecules or genetic absence of gingival SCCs (gSCCs) increases the bacterial load, reduces bacterial diversity, and renders the microbiota more pathogenic, leading to greater alveolar bone loss. Topical treatment with bitter denatonium to activate gSCCs upregulates the expression of antimicrobial peptides and ameliorates ligature-induced periodontitis in wild-type but not in Gnat3-/- mice. We conclude that gSCCs may provide a promising target for treating periodontitis by harnessing innate immunity to regulate the oral microbiome.


Assuntos
Células Quimiorreceptoras/imunologia , Gengiva/imunologia , Imunidade Inata , Microbiota/imunologia , Periodontite/imunologia , Animais , Células Quimiorreceptoras/metabolismo , Modelos Animais de Doenças , Feminino , Gengiva/citologia , Gengiva/microbiologia , Células HEK293 , Proteínas Heterotriméricas de Ligação ao GTP/genética , Proteínas Heterotriméricas de Ligação ao GTP/metabolismo , Humanos , Masculino , Camundongos , Camundongos Knockout , Mucosa Bucal/citologia , Mucosa Bucal/imunologia , Mucosa Bucal/metabolismo , Periodontite/microbiologia , Fosfolipase C beta/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Transdução de Sinais/imunologia , Canais de Cátion TRPM/metabolismo
7.
Eur. j. anat ; 23(5): 383-388, sept. 2019. ilus, graf, tab
Artigo em Inglês | IBECS | ID: ibc-183869

RESUMO

Langerhans cells (LCs) are the most effective antigen presenting cells against foreign bodies and carcinogens. Since the oral cavity is a portal of entry for these antigens, the aim of this study was to morphologically classify CD1a+ LCs, quantify them in the normal and malignant buccal mucosa, and evaluate their relation to the age of patients. Healthy buccal mucosal samples collected from 16 patients undergoing reconstructive operation, and malignant samples obtained from 15 patients undergoing radical oncological resection. were processed for immunohistochemistry four- to fivemicron thick sections were stained with CD1a antibody (CD1a). At 40X magnification, CD1a+ LCs were morphologically classified and quantified manually for a 25mm length of basement membrane using Cellsens image analysing software and the data was analysed. Two categories of CD1a+ LCs were identified in the normal and malignant buccal mucosa a) typical dendritic LCs and b) non-dendritic LCs (a new entity). Non-dendritic LCs were of significantly higher number compared to the typical dendritic LCs in the normal tissues (p -0.001). In the malignant group, the non-dendritic CD1a+ LCs were significantly fewer in number (p0.004), when compared to the normal group. Nondendritic LCs were also significantly fewer (p0.026) in patients over 60 years of age. This is the first report of non-dendritic Langerhans cells in normal buccal mucosa and malignant buccal mucosa using the CD1a marker. The significantly higher number of these cells in normal tissues and younger individuals supports their role as accessory antigen presenting cells


No disponible


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Células de Langerhans/patologia , Mucosa Bucal/anatomia & histologia , Mucosa Bucal/patologia , Células de Langerhans/efeitos dos fármacos , Mucosa Bucal/citologia , Antígenos CD1
8.
Niger J Clin Pract ; 22(8): 1041-1048, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31417045

RESUMO

Background: X-rays are potential mutagenic agents that can cause both the gene mutations and chromosomal aberrations. Aims: In this study, the micronucleus (MN) test and the comet assay methods are implemented in order to observe the damage that can occur in the cell nucleus and in the structure of DNA of the patients who underwent a panoramic examination. Methods and Materials: In our study, buccal mucosa swabs were obtained just before the radiography and 2 weeks after the radiography from 30 volunteer patients who had to take radiographs due to dental diagnosis. Changes in the nuclei of 1,000 cells of each swab sample had been counted under a light microscope and recorded. Besides, 100 cells of each other swab samples were analyzed by the comet assay. Comet assay parameters namely tail length and percentage of DNA in tail, which indicate the level of DNA damage were analyzed and compared in both groups. Statistical analysis was performed by using the Statistical Package for the Social Sciences (Version 21). Results: In our study, the results of percentage of DNA in tail and tail length before and after X-ray exposure were statistically significant (P < 0.001). Likewise, increase in the MN frequency observed in buccal mucosa cells after X-ray exposure was found significant (P < 0.001). Conclusions: As a result, panoramic radiographs taken during dental diagnosis and treatment cause cytotoxicity and DNA damage in oral mucosal cells. Panoramic radiographs should be applied only when necessary, using an accurate radiographic technique and radioprotection criteria.


Assuntos
Núcleo Celular/efeitos da radiação , Dano ao DNA/efeitos da radiação , Mucosa Bucal/efeitos da radiação , Radiografia Panorâmica/efeitos adversos , Ensaio Cometa , Feminino , Humanos , Masculino , Testes para Micronúcleos , Mucosa Bucal/citologia , Testes de Mutagenicidade
9.
Forensic Sci Int Genet ; 43: 102142, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31437782

RESUMO

Small variations between haplotypes detected in different tissues from the same individual have been previously described. These differences complicate the interpretation of mtDNA results in real forensic casework. mtDNA haplotypes detected in hair strands collected at the crime scene have to be frequently compared with haplotypes of reference samples (buccal swabs) from victims or suspects. Nucleotide position 16093 is a well-known hot spot where differences can accumulate between different tissues of the same individual. Intra individual variation was also detected at positions 16182 and 16183 in haplotypes showing an uninterrupted HV1 poly-C stretch (with 16189C). In order to better characterize the type of variation in these positions between buccal cells and hair strands from the same individual, we have performed Sanger sequencing in 25-28 hair strands (411 in total) from 15 individuals showing either an uninterrupted HV1 polyC-stretch (16189C) or 16093C/Y in their buccal cells. The results have been evaluated by also taking into account our previous results published in [19]. We have found that no variation among hair strands was detected in individuals showing T16093 in buccal cells, while variation in hair strands (T16093, 16093C and 16093Y) were detected in individuals showing 16093C or 16093Y in buccal cells. Regarding nucleotide positions 16182 and 16183 in combination with an uninterrupted polyC-stretch, no variation was detected in hairs from individuals showing A16182 16183C in their buccal cells. In contrast, individuals A16182 A16183 showed hair strands with A16182 16183 M and A16182 16183C. And finally, individuals with 16182C 16183C showed some variation in a small amount of their hair strands (some hairs with 16182 M 16183C). These results can be relevant for forensic practitioners when comparing reference samples with hair strands, which is the type of sample most tested by using mtDNA analysis in forensic casework.


Assuntos
DNA Mitocondrial/genética , Cabelo/química , Polimorfismo Genético , Análise de Sequência de DNA , Sequência de Bases , Células Epiteliais/química , Humanos , Mucosa Bucal/citologia , Reação em Cadeia da Polimerase
10.
Cornea ; 38(10): 1273-1279, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31356413

RESUMO

PURPOSE: To evaluate the outcomes of autologous cultivated oral mucosal epithelial transplantation (COMET) in ocular surface reconstructive procedures. METHODS: Twenty-five eyes of 24 patients who underwent COMET for ocular surface reconstruction were studied retrospectively from April 2011 to March 2014. Oral mucosal biopsy of 2 mm was harvested from all patients and cultured on amniotic membrane for 14 days. COMET was performed as a means to achieve corneal epithelization (group 1) and for fornix reconstruction (group 2). The extent of epithelization achieved (group 1) and the depth of the fornix reconstructed (group 2) were the primary outcome measures. RESULTS: Six eyes of 5 patients [2 chemical injury (CI), 3 Stevens-Johnson Syndrome (SJS)] underwent COMET for nonhealing of an epithelial defect within a month after insult or had a nonhealing defect since insult despite maximal medical/surgical therapy. Group 2 included 19 eyes of 19 patients (10 CI, 8 SJS, 1 ocular cicatricial pemphigoid) which underwent COMET for fornix reconstruction. Postsurgery, the cornea was fully epithelized in 66.67% of the eyes (n = 4) and partially epithelized in 33.33% of the eyes (n = 2). The overall improvement in surface epithelization was statistically significant (P = 0.046). Reconstruction of an anatomically deep fornix was achieved in 57% of the eyes (4 CI, 6 SJS and 1 ocular cicatricial pemphigoid). The fornix was partially formed in 26% of the eyes (4 CI and 1 SJS). The overall improvement in fornix reconstruction was considered statistically significant (P = 0.024). The follow-up ranged from 1 to 5 to 27 months with a mean follow-up of 18 months SD ±8.9. CONCLUSIONS: COMET, by providing an alternate source of epithelium, aids in faster epithelization and thus can be considered as an option in management of severe grade CI or SJS in the acute stage as well as in fornix reconstructive procedures in chronic stage of ocular surface disorders.


Assuntos
Córnea/cirurgia , Doenças da Córnea/cirurgia , Células Epiteliais/transplante , Mucosa Bucal/transplante , Procedimentos Cirúrgicos Oftalmológicos/métodos , Procedimentos Cirúrgicos Reconstrutivos/métodos , Acuidade Visual , Adolescente , Adulto , Idoso , Células Cultivadas , Criança , Pré-Escolar , Córnea/patologia , Doenças da Córnea/diagnóstico , Células Epiteliais/citologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/citologia , Reepitelização , Estudos Retrospectivos , Transplante Autólogo , Adulto Jovem
11.
Cells ; 8(7)2019 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-31336838

RESUMO

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.


Assuntos
Fibroblastos , Queratinócitos , Mucosa Bucal , Óxido Nítrico/metabolismo , Palato , beta-Defensinas/metabolismo , Biofilmes , Candida albicans/fisiologia , Candidíase/imunologia , Candidíase/microbiologia , Linhagem Celular , Fibroblastos/citologia , Fibroblastos/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Queratinócitos/citologia , Queratinócitos/metabolismo , Mucosa Bucal/citologia , Mucosa Bucal/metabolismo , Palato/citologia , Palato/metabolismo
12.
J Biol Regul Homeost Agents ; 33(3): 695-706, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31184088

RESUMO

Lipids are an alternative energy source for cells and provide structural integrity in cell membrane and their metabolism is regulated with the use of different pathways, such as integrin signalling, oxidative stress, mechanical stress, and pH changes. All of those processes take place in the oral mucosa which is subject to different environmental impacts. In this study, porcine buccal pouch mucosal cells (pBPMCs) were used during long-term primary in vitro culture. The cultured cells were collected at 7, 15 and 30 days of IVC and subsequently transferred to RNA isolation. In the results of the following microarray analysis, we analyzed the genes detected, belonging to ontology groups, such as "cellular lipid metabolic process", "response to lipid" and "response to lipopolysaccharides. All of the genes involved in these ontological groups were expressed at higher levels at 7 days of IVC and substantially decreased in expression at days 15 and 30 of primary culture. We observed new genes, which may be recognized as markers in regulation of lipid metabolism in mucosal cells in vitro. The results suggested that the biochemical mechanism-involved lipids were accompanied by increased enzymatic activation and synthesis of crucial growth factors reaching high activity at day 7 of culture, which is also well documented as a stage of tissue regeneration period within oral mucosa. Therefore, this "biochemical fingerprint" may be an additional checkpoint of the integrity, resistance and easy adaptability of oral tissues, which are important conditions of success in tissue engineering and grafting for tissue reconstruction.


Assuntos
Expressão Gênica , Metabolismo dos Lipídeos , Lipopolissacarídeos , Mucosa Bucal/citologia , Animais , Células Cultivadas , Bochecha , Análise de Sequência com Séries de Oligonucleotídeos , Cultura Primária de Células , Suínos
13.
Middle East Afr J Ophthalmol ; 26(1): 23-26, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31114120

RESUMO

PURPOSE: The purpose of this study is to analyze the ocular surface microbial flora in patients with chronic limbal stem cell deficiency (LSCD) due to Stevens-Johnson Syndrome (SJS) and ocular chemical injury undergoing cultivated oral mucosal epithelial transplantation (COMET). METHODS: Patients of SJS and chemical injury who had bilateral total LSCD planned for COMET were studied. Conjunctival swab was taken before surgery. Parameters evaluated were organism cultured, sensitivity pattern, frequency of positive culture, and clinical impact on management strategy. RESULTS: Thirteen patients were included in which nine were males and four females. All patients had positive conjunctival swab culture. Most common organism isolated was Staphylococcus epidermidis, followed by Staphylococcus aureus and Pseudomonas aeruginosa. The staphylococcal species isolated were sensitive to all the conventional antibiotics while Pseudomonas cultured showed resistance to cefuroxime, ceftriaxone, and ceftazidime. Repeat conjunctival swab sent after a week of topical antibiotic therapy yielded positive culture of the same organism twice in 25% (3/12), thrice in 58.3% (7/12), and four times in 16.6% (2/12) of the patients. One patient had a polymicrobial flora with positive yield of S. aureus (thrice), S. epidermidis (twice), and P. aeruginosa (twice) in consecutive conjunctival swab culture in the absence of clinical infection. Two patients with persistent positive cultures had to undergo repeat oral mucosal harvesting as the transplantation of the cultivated explants had to be deferred. CONCLUSION: Ocular surface in LSCD patients yielded pathogenic organisms on culture. Poor ocular surface with absent tear film could be the contributing factors. It is important to perform the conjunctival swab culture before COMET surgery.


Assuntos
Bactérias/isolamento & purificação , Túnica Conjuntiva/microbiologia , Doenças da Córnea/cirurgia , Células Epiteliais/transplante , Limbo da Córnea/patologia , Mucosa Bucal/citologia , Células-Tronco/patologia , Adolescente , Adulto , Técnicas Bacteriológicas , Queimaduras Químicas/cirurgia , Células Cultivadas , Criança , Doença Crônica , Queimaduras Oculares/induzido quimicamente , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Síndrome de Stevens-Johnson/cirurgia , Adulto Jovem
14.
Klin Lab Diagn ; 64(4): 229-233, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31108036

RESUMO

The Russian northern regions development is associated with the extraction and processing of natural resources, which leads to environmental pollution and makes the task of sanitary-hygienic monitoring relevant. Buccal micronucleus cytome assay is one of the toxicological methods for human population studies. Studies on the inter-latitudinal comparison of Buccal micronucleus cytome assay are lacking, therefore there are difficulties in comparing the results obtained in the Russian Arctic with data from more southern regions. The aim of this study is to compare cytogenetic abnormalities in the buccal epithelium in two groups of older schoolchildren living in high and middle latitudes. The study was conducted in the city of Apatity (Murmansk region, 67 ° 34'03 ″ N, 33 ° 23'36 ″ E) and the city of Serpukhov (Moscow region, 54 ° 54 '56 ″ N, 37 ° 24 '40 "E). A total of 61 children were examined: 41 children from the Apatity and 20 children from the Serpukhov (16-18 years old). The Buccal micronucleus cytome assay was carried out according to an international protocol. It was shown that the average frequency values of the cells with micronucleus in the comparison groups of schoolchildren living in high and middle latitudes did not significantly differ and did not exceed the values for the average population norm. The frequency of cells with nuclear buds and two nucleus was significantly higher in the group of schoolchildren living in middle latitudes, which, in turn, is compensated by a higher rate of elimination of cells with impaired. Therefore, when comparing Buccal micronucleus cytome assay data, it is quite possible not to take into account the breadth of the studied groups.


Assuntos
Células Epiteliais/citologia , Mucosa Bucal/citologia , Adolescente , Regiões Árticas , Humanos , Testes para Micronúcleos , Moscou , Federação Russa
15.
Environ Sci Pollut Res Int ; 26(19): 19676-19683, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31079294

RESUMO

Since many different pesticides have been used occupationally, there have been inconsistent results regarding DNA damages among greenhouse workers. Thus, the aim of the study is to evaluate DNA damages, cell death, and chromosomal instability by using the buccal micronucleus cytome (BMcyt) assay in greenhouse workers and to compare those with a non-exposed group. The BMcyt assay was applied to the exfoliated buccal cell samples collected from 66 pesticide-exposed and 50 non-exposed individuals. We evaluated the frequency of micronucleus (MN), nuclear bud (NBUD), binucleated (BN) cells, and karyolitic (KL), pyknotic (PY), and karyorrhectic (KH) cells. The results showed that the MN, BN, PY, and KH frequencies of the pesticide-exposed group were significantly higher than those of the controls (P Ë‚ 0.05, P Ë‚ 0.05, P Ë‚ 0.01, and P Ë‚ 0.05, respectively). We observed that the MN, BN, PY, and KH frequencies in the autumn were statistically different compared with those in the control group (P = 0.037 for MN, P = 0.001 for BN, P = 0.016 for PY, and P = 0.033 for KH). The same comparison was done in the spring for the control, and there was a statistically significant difference for MN (P = 0.046) and PY (P = 0.014). We can conclude that pesticide exposure in greenhouse workers was one of the factors that altered DNA damages, cell death, and chromosomal instability in oral mucosa cells.


Assuntos
Instabilidade Cromossômica/efeitos dos fármacos , Dano ao DNA , Monitoramento Ambiental/métodos , Fazendeiros , Mucosa Bucal/efeitos dos fármacos , Exposição Ocupacional/análise , Praguicidas/toxicidade , Adulto , Morte Celular/efeitos dos fármacos , Feminino , Humanos , Masculino , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Testes para Micronúcleos , Mucosa Bucal/citologia , Exposição Ocupacional/efeitos adversos , Turquia
16.
Graefes Arch Clin Exp Ophthalmol ; 257(6): 1253-1263, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31004182

RESUMO

PURPOSE: The purpose of this study was to compare the efficacy of allogeneic cultured limbal epithelial transplantation (ACLET) and cultivated oral mucosal epithelial transplantation (COMET) in treating total limbal stem cell deficiency (LSCD). METHODS: In this retrospective cohort study, 73 patients (76 eyes) with total LSCD, including 41 patients (42 eyes) treated with ACLET and 32 patients (34 eyes) receiving COMET, were evaluated. The age, gender and injury cause of all patients were recorded. RESULTS: The mean follow-up was 23.3 ± 9.9 months in the ACLET group and 16.1 ± 5.8 months in the COMET group. A higher incidence of persistent epithelial defect was observed after COMET (P = 0.023). The overall ocular surface grading scores were all lower in the ACLET group than in the COMET group at 3, 6, and 12 months after surgery and the last follow-up. Kaplan-Meier survival curve analysis demonstrated a significantly higher success rate of ACLET (71.4%), compared with that of COMET (52.9%; P = 0.043). The risk of graft failure was higher in patients with entropion and trichiasis, incomplete eyelid closure and treated with COMET. The graft failure risk rate after COMET was 3.5 times higher than that of ACLET. CONCLUSIONS: For total LSCD patients, ACLET should be prioritized, since limbal epithelial cells have better ability to maintain corneal epithelial integrity and ocular surface stability and benefit the ocular surface when compared with oral mucosal epithelial cells. Preoperative and postoperative eyelid abnormalities should be corrected as early as possible.


Assuntos
Doenças da Córnea/cirurgia , Limbo da Córnea/patologia , Mucosa Bucal/transplante , Transplante de Células-Tronco/métodos , Acuidade Visual , Adolescente , Adulto , Células Cultivadas , Doenças da Córnea/patologia , Células Epiteliais/citologia , Células Epiteliais/transplante , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/citologia , Estudos Retrospectivos , Transplante Autólogo , Resultado do Tratamento , Adulto Jovem
17.
Forensic Sci Int Genet ; 41: 42-49, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30954892

RESUMO

We report the successful separation of sperm cells from a relevant composition of mock sexual assault samples using a novel acoustic differential extraction (ADE) technology. A multi-layer microfluidic device fabricated in a non-photolithographic process from glass and polydimethylsiloxane (PDMS) was capable of interfacing with custom-built instrumentation to exploit a standing acoustic wave for the trapping of individual sperm cells in a sample containing an abundance of epithelial cells. Samples were generated from buccal and vaginal swabs to mimic post-coital vaginal swabs, and processed through the ADE system followed by DNA extraction of the captured cells with amplification of DNA using a custom short tandem repeat (STR) chemistry. The prototype acoustic trapping technology was fully capable of isolating intact sperm cells from mock samples with disparate masses of male and female DNA. Other biological components were evaluated for adverse effects on sperm cell trapping, including blood, yeast, and bacteria (E. coli), and these had negligible effects on observed sperm cell trapping. Finally, we demonstrate the successful capture of sperm cells from mock samples containing a 40-fold excess in female epithelial cells over sperm cells. The effectiveness of sperm cell purification was ascertained with polymerase chain reaction (PCR) amplification of STR loci from the male fraction post separation with an 18-plex amplification kit, which resulted in male-only profiles.


Assuntos
Impressões Digitais de DNA , Técnicas Analíticas Microfluídicas , Sêmen/citologia , Delitos Sexuais , Manejo de Espécimes , Separação Celular , Feminino , Humanos , Masculino , Repetições de Microssatélites , Mucosa Bucal/citologia , Reação em Cadeia da Polimerase , Vagina/citologia
18.
Cell Mol Biol Lett ; 24: 3, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30988673

RESUMO

Background: Autologous cultivated oral mucosal epithelial transplantation (COMET) is an important treatment for limbal stem cell deficiency. However, peripheral corneal neovascularization after surgery hinders its application. This study aims to employ a culture system using allogenic limbal niche cells (LNCs) instead of mouse-derived 3T3 cells as a feeder layer that could relieve postoperative neovascularization. Methods: Rat oral mucosal epithelial cells (OMECs) were co-cultured with rat LNCs or 3T3 cells. Cultivated oral mucosal epithelial cells (COMECs) of different culture systems were identified by hematoxylin and eosin staining and immunocytochemistry. The expression levels of the angiogenesis-related factors were analyzed by RT-qPCR and western blotting/ELISA. Angiogenic potential was reconfirmed by cell viability and tube formation assays with human umbilical vein endothelial cells (HUVECs). Results: COMECs were obtained from both culture systems successfully. Immunocytochemistry showed approximately equal percentages of positive staining cells for p63α (p = 0.9177), ABCG2 (p = 0.526), Ki67 (p = 0.0987), and CK3 (p = 0.4000) in COMECs of different groups. RT-qPCR and western blotting/ELISA showed that COMECs of the LNC group expressed a significantly lower amount of basic fibroblast growth factor (bFGF) (p = 0.0038 for RT-qPCR, p = 0.0026 for western blotting) but more pigment epithelium-derived factor (PEDF) (p = 0.0172 for RT-qPCR, p = 0.0253 for western blotting) and soluble fms-like tyrosine kinase-1 (sFlt-1) (p < 0.0001 for RT-qPCR, p = 0.0064 for ELISA) than the COMECs of the 3T3 group. Furthermore, compared with COMECs of the 3T3 group, COMECs of the LNC group could reduce the viability (p = 0.0002) and tube formation (p = 0.0002) of HUVECs. Conclusions: LNCs could substitute 3T3 cells for expanding OMECs in vitro, and the COMECs obtained in this system are less likely to induce postsurgical neovascularization, which provides an alternative option for an ex vivo culture system and promotes the application of COMET.


Assuntos
Células Epiteliais/citologia , Limbo da Córnea/citologia , Mucosa Bucal/citologia , Neovascularização Fisiológica , Nicho de Células-Tronco , Células 3T3 , Animais , Biomarcadores/metabolismo , Proliferação de Células , Forma Celular , Sobrevivência Celular , Células Cultivadas , Técnicas de Cocultura , Células Epiteliais/metabolismo , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Camundongos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley
19.
Ecotoxicol Environ Saf ; 179: 135-142, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31035247

RESUMO

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.


Assuntos
Misturas Complexas/toxicidade , Dano ao DNA , Fertilizantes/toxicidade , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Exposição Ocupacional/efeitos adversos , Praguicidas/toxicidade , Polimorfismo Genético , Adulto , Apoptose/efeitos dos fármacos , Arildialquilfosfatase/efeitos dos fármacos , Brasil , Simulação por Computador , Células Epiteliais/efeitos dos fármacos , Fazendeiros , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/citologia , Exposição Ocupacional/análise , Proteína 1 Complementadora Cruzada de Reparo de Raio-X/genética
20.
Int J Oncol ; 54(5): 1567-1578, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30896790

RESUMO

Emerging evidence indicates that mesenchymal stem cells (MSCs) serve an indispensable role in the tumor microenvironment. However, whether MSCs participate in the development of oral carcinogenesis remains unclear. The present study isolated MSCs from clinical tissues and investigated the differences of MSCs derived from normal oral mucosa (N­MSC), oral leukoplakia with dysplasia (LK­MSC) and oral carcinoma (Ca­MSC). The results revealed that the LK­MSCs exhibited reduced proliferation and migration, compared with the N­MSCs and Ca­MSCs. Furthermore, it was demonstrated that the exosomes secreted by LK­MSCs have significant roles in promoting proliferation, migration and invasion in vitro, which was similar to the Ca­MSC­derived exosomes. The promoting effect was also demonstrated in a 3D coculture model. When the secretion of exosomes was blocked, the promoting effect of LK­MSCs was reversed. Based on a microarray analysis of MSC­derived exosomes, microRNA­8485 (miR­8485) was identified to be ectopically expressed. The exosomal miR­8485 was capable of promoting the proliferation, migration and invasion of tumor cells. Therefore, the present study highlights the significance of MSC­derived exosomes and exosomal miR­8485 in premalignant lesions and carcinogenesis. Intervention with the secretion of MSC­derived­exosomes may be an innovative strategy to retard the carcinogenesis.


Assuntos
Exossomos/patologia , Leucoplasia Oral/patologia , MicroRNAs/genética , Mucosa Bucal/citologia , Neoplasias Bucais/patologia , Adulto , Idoso , Diferenciação Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura/métodos , Exossomos/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Leucoplasia Oral/genética , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Mucosa Bucal/metabolismo , Mucosa Bucal/patologia , Neoplasias Bucais/genética , Microambiente Tumoral , Regulação para Cima
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