Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.838
Filtrar
1.
In Vitro Cell Dev Biol Anim ; 57(2): 148-159, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33447968

RESUMO

Three-dimensional, organotypic models of the oral mucosa have been developed to study a wide variety of phenomena occurring in the oral cavity. Although a number of models have been developed in academic research labs, only a few models have been commercialized. Models from academic groups offer a broader range of phenotypes while the commercial models are more focused on the oral and gingival mucosa. The commercialized models are manufactured under highly controlled conditions and meet the requirements of quality standards, which leads to high levels of reproducibility. These in vitro models have been used to evaluate the irritancy of oral care products such as toothpastes, mouthwashes, and mucoadhesives. The effects of cigarette smoke on oral cavity tissues have been studied and compared to those of e-cigarettes. Oral tissue models have facilitated investigation of the mechanisms of oral mucositis and oral candidiasis and have been used to examine transbuccal drug delivery rates and the absorption of nanoparticles. Infection studies have investigated the effects of HIV-1 along with the effects of commensal and pathogenic bacteria. More recently, a differentiated oral tissue model has been shown to express the ACE2 receptor, which is known to be important for the receptor-mediated entry of the SARS-CoV-2 coronavirus into human cells and tissues. Hence, oral mucosal models may find application in determining whether viral infection of the oral mucosa is possible and whether such infection has implications vis-a-vis the current COVID-19 pandemic. As is apparent, these models are used in a broad variety of applications and often offer advantages versus animal models in terms of reproducibility, avoiding species extrapolation, and the ethical concerns related to human and animal experimentation. The goals of this paper are to review commercially available models of the human buccal and gingival mucosa and highlight their use to gain a better understanding of a broad range of phenomena affecting tissues in the oral cavity.


Assuntos
Materiais Dentários/efeitos adversos , Infecções , Mucosa Bucal/citologia , Mucosa Bucal/virologia , Técnicas de Cultura de Tecidos/métodos , /transmissão , Sistemas de Liberação de Medicamentos/efeitos adversos , Sistemas de Liberação de Medicamentos/métodos , Humanos , Infecções/microbiologia , Infecções/virologia , Boca , Mucosa Bucal/microbiologia , Mucosa Bucal/patologia , Controle de Qualidade , Técnicas de Cultura de Tecidos/instrumentação , Engenharia Tecidual , Tabaco , Cremes Dentais/efeitos adversos , Raios Ultravioleta/efeitos adversos
2.
Nanotoxicology ; 15(2): 223-237, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33373530

RESUMO

The available biomonitoring studies on workers producing/handling nanomaterials (NMs) focused on potential effects on respiratory, immune and cardio-vascular system. Aim of this study was to identify a panel of sensitive biomarkers and suitable biological matrices to evaluate particularly genotoxic and oxidative effects induced on workers unintentionally exposed to graphene or silica nanoparticles during the production process. These nanomaterials have been chosen for 'NanoKey' project, integrating the workplace exposure assessment (reported in part I) with the biomonitoring of exposed workers reported in the present work. Simultaneously to workplace exposure characterization, we monitored the workers using: Buccal Micronucleus Cytome (BMCyt) assay, fpg-comet test (lymphocytes), oxidized DNA bases 8-oxoGua, 8-oxoGuo and 8-oxodGuo measurements (urine), analysis of oxidative stress biomarkers in exhaled breath condensate (EBC), FENO measurement and cytokines release detection (serum). Since buccal cells are among the main targets of NM occupational exposure, particular attention was posed to the BMCyt assay that represents a noninvasive assay. This pilot study, performed on 12 workers vs.11 controls, demonstrates that BMCyt and fpg-comet assays are the most sensitive biomarkers of early, still reparable, genotoxic and oxidative effects. The findings suggest that these biomarkers could represent useful tools for the biomonitoring of workers exposed to nanoparticles, but they need to be confirmed on a high number of subjects. However, such biomarkers don't discriminate the effects of NM from those due to other chemicals used in the NM production process. Therefore, they could be suitable for the biomonitoring of workers exposed to complex scenario, including nanoparticles exposure.


Assuntos
Dano ao DNA , Grafite/toxicidade , Mucosa Bucal/efeitos dos fármacos , Nanopartículas/toxicidade , Exposição Ocupacional/efeitos adversos , Estresse Oxidativo/efeitos dos fármacos , Dióxido de Silício/toxicidade , Adulto , Biomarcadores/metabolismo , Células Cultivadas , Ensaio Cometa , Citocinas/metabolismo , Feminino , Grafite/administração & dosagem , Humanos , Inflamação , Masculino , Testes para Micronúcleos , Mucosa Bucal/citologia , Mucosa Bucal/metabolismo , Nanopartículas/administração & dosagem , Exposição Ocupacional/análise , Oxirredução , Estresse Oxidativo/genética , Estresse Oxidativo/imunologia , Projetos Piloto , Dióxido de Silício/administração & dosagem , Local de Trabalho/normas
3.
Artigo em Inglês | MEDLINE | ID: mdl-33321868

RESUMO

Many subjects perceive venous blood collection as too invasive, and thus moving to better-accepted procedures for leukocytes collection might be crucial in human biomonitoring studies (e.g., biomonitoring of occupational or residential exposure to genotoxins) management. In this context, primary DNA damage was assessed in buccal lymphocytes (BLs), fresh whole venous, and capillary blood leukocytes, and compared with that in peripheral blood lymphocytes (PBLs)-the most frequently used cells-in 15 young subjects. Mouthwashes were collected after the volunteers rinsed their mouths with normal saline, and BLs were isolated by density gradient centrifugation. Blood samples were collected by venipuncture or by lancet. Anthropometric and lifestyle information was obtained by the administration of a structured questionnaire. As shown in the Bland-Altman plots, the level of agreement between BLs and PBLs lied within the accepted range, we thus enrolled a wider population (n = 54) to assess baseline DNA damage in BLs. In these cells, mean values of tail length (µm), tail intensity (%), and tail moment were 25.7 ± 0.9, 6.7 ± 0.4 and 1.0 ± 0.1, respectively. No significant association was observed between sex and smoking habit with any of the DNA damage parameters. Conversely, underweight subjects displayed significantly higher genomic instability compared with normal weight group (p < 0.05). In conclusion, we successfully managed to set up and update a non-invasive and well-accepted procedure for the isolation of BLs from saliva that could be useful in upcoming biomonitoring studies.


Assuntos
Monitoramento Biológico , Ensaio Cometa , Dano ao DNA , Linfócitos , Monitoramento Biológico/métodos , Feminino , Humanos , Leucócitos/patologia , Linfócitos/patologia , Masculino , Mucosa Bucal/citologia
5.
Ecotoxicol Environ Saf ; 203: 110989, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32888600

RESUMO

AIM: The city of L'Aquila (central Italy) was hit by a strong earthquake in 2009 that caused the collapse of several buildings, deaths and injured people. In the following years, a great number of building sites were activated, building workers resulted intensely exposed and represent a relevant target for research on environmental mutagenesis and epidemiological surveillance. Cells of buccal mucosa are considered an appropriate site for early detecting of cytogenetic damage, since it represents the first barrier in inhalation or ingestion and can metabolize carcinogenic agents into reactive chemicals. Our study is aimed 1) at comparing the early genotoxic damage as measured by the buccal mucosa micronucleus test in two subgroups of workers defined by different occupational exposure and 2) at evaluating possible confounding variables such as lifestyle factors. METHODS AND RESULTS: A cross-sectional study was conducted in L'Aquila, on 24 outdoor workers (OWs) highly exposed on the construction sites and 26 indoor workers (IWs), all subjected to the compulsory occupational surveillance system, in the period 2017-2018. Buccal cells samples were collected and, based on the Micronucleus test, the exfoliated cells were classified in respect of nuclear changes observed. Moreover, a self-report questionnaire composed of 84 items, was administered to the workers. RESULTS: Significant differences were observed between Exp+ (OWs) and Exp- (IWs) in the number of the analyzed cells (expressed as mean value out of 1000 cells): respectively 954.46 vs 990.06 normal cells, (p < 0.001); 19.79 vs 4.95 micronucleated cells, as marker of chromosomal damage (p < 0.001); 13.93 vs 8.96 binucleated cells, as marker of failed cytokinesis (p < 0.001); 2.09 vs 1.18 karyolytic cells, as marker of cell death and damaged DNA (p < 0.05). According with a multivariate regression analysis, in addition to the job exposure (OW vs IW, beta = 12.221, p < 0.001), the only variable independently associated with an increase in Micronuclei (MNs) is the smoking habit (OWs vs IWs, beta = 6.683, p < 0.001) which, even if not associated with dust exposure, worsens cell integrity. Moreover, this worsening effect is weaker in workers not exposed to the site dust (moderation effect). Within social demographic factors, the high educational level only apparently seems to affect MNs number: even if unbalanced in favor of IWs vs OWs, this variable resulted a confounder, since its effect disappears when the interaction between these two factors is considered, because it is a covariate of smoking habit as well as of the job condition. CONCLUSION: Despite some limitation, our findings clearly confirm the role of occupational exposure as a marker of cytogenetic damage associated with MNs number in construction workers. Moreover, smoking status appears as the only other investigated factor independently associated to the outcome. The statistical model, in addition, highlights possible moderation and confounding effects, such as interaction between smoking and occupational exposure and the unbalanced school education level in workers. Micronucleus test in exfoliated buccal cells would be considered a suitable method for studying the early genotoxic damage in the construction occupational setting as well as in evaluating the efficacy of preventive practices.


Assuntos
Indústria da Construção , Dano ao DNA , Poeira , Mucosa Bucal/citologia , Exposição Ocupacional/efeitos adversos , Adulto , Estudos Transversais , Humanos , Itália , Masculino , Testes para Micronúcleos , Pessoa de Meia-Idade , Adulto Jovem
6.
Int J Mol Sci ; 21(18)2020 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-32957696

RESUMO

At present, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has quickly become a health emergency because no specifics vaccines or drugs, at this moment, are available. Recent studies have shown that the transplantation of mesenchymal stem cells (MSCs) into Coronavirus Disease 2019 (COVID-19) patients could represent a promising strategy for the development of new therapeutic methods. We speculate and suggest that the secretome of human Oral Tissue Stem Cells (hOTSCs), for their immunomodulatory and anti-inflammatory specific properties, could exert beneficial effects on the COVID-19 patients through an innovative aerosolisation technique. This non-invasive technique can offer multiple advantages in prophylaxis, as well as the prevention and treatment of severe epidemic respiratory syndrome with minimum risk and optimal therapeutic effects. This has the potential to create a novel pathway towards immunomodulatory therapy for the treatment of COVID-19 positive patients.


Assuntos
Infecções por Coronavirus/tratamento farmacológico , Fatores Imunológicos/uso terapêutico , Células-Tronco Mesenquimais/metabolismo , Mucosa Bucal/citologia , Pneumonia Viral/tratamento farmacológico , Proteoma/uso terapêutico , Humanos , Fatores Imunológicos/metabolismo , Pandemias , Proteoma/metabolismo , Via Secretória
7.
Toxicol Lett ; 333: 242-250, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32841739

RESUMO

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.


Assuntos
Bioensaio/normas , Núcleo Celular/genética , Células Epiteliais/ultraestrutura , Laboratórios/normas , Micronúcleos com Defeito Cromossômico , Testes para Micronúcleos/normas , Mucosa Bucal/citologia , Adolescente , Adulto , Bioensaio/métodos , Brasil , Morte Celular/genética , Núcleo Celular/ultraestrutura , Dano ao DNA , Feminino , Humanos , Masculino , Micronúcleos com Defeito Cromossômico/estatística & dados numéricos , Testes para Micronúcleos/métodos , Pessoa de Meia-Idade , Mucosa Bucal/ultraestrutura , Adulto Jovem
9.
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
10.
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
11.
J. oral res. (Impresa) ; 9(2): 104-110, abr. 30, 2020. ilus, graf, tab
Artigo em Inglês | LILACS | ID: biblio-1151903

RESUMO

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.


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Adulto Jovem , Testes para Micronúcleos , Micronúcleos com Defeito Cromossômico , Uso de Tabaco/efeitos adversos , Fumar Tabaco/efeitos adversos , Mucosa Bucal/citologia , Técnicas In Vitro , Aberrações Cromossômicas , não Fumantes , Índia
12.
Int J Mol Sci ; 21(4)2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-32075221

RESUMO

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.


Assuntos
Mucosa Bucal/crescimento & desenvolvimento , Osteogênese/genética , Ligamento Periodontal/crescimento & desenvolvimento , Proteínas/química , Células-Tronco/citologia , Regeneração Óssea/genética , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/genética , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Cemento Dentário/metabolismo , Durapatita/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Glicogênio Sintase Quinase 3 beta/genética , Humanos , Sialoproteína de Ligação à Integrina/genética , Mucosa Bucal/citologia , Mucosa Bucal/metabolismo , Osteoblastos/metabolismo , Osteocalcina/genética , Peptídeos/química , Peptídeos/genética , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Estrutura Secundária de Proteína , Proteínas/genética , Proteínas/ultraestrutura , Fator de Transcrição Sp7/genética , Células-Tronco/metabolismo , Via de Sinalização Wnt/genética
13.
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
14.
Int J Mol Sci ; 21(2)2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31936462

RESUMO

The corneal surface is an essential organ necessary for vision, and its clarity must be maintained. The corneal epithelium is renewed by limbal stem cells, located in the limbus and in palisades of Vogt. Palisades of Vogt maintain the clearness of the corneal epithelium by blocking the growth of conjunctival epithelium and the invasion of blood vessels over the cornea. The limbal region can be damaged by chemical burns, physical damage (e.g., by contact lenses), congenital disease, chronic inflammation, or limbal surgeries. The degree of limbus damage is associated with the degree of limbal stem cells deficiency (partial or total). For a long time, the only treatment to restore vision was grafting part of the healthy cornea from the other eye of the patient or by transplanting a cornea from cadavers. The regenerative medicine and stem cell therapies have been applied to restore normal vision using different methodologies. The source of stem cells varies from embryonic stem cells, mesenchymal stem cells, to induced pluripotent stem cells. This review focuses on the use of oral mucosa epithelial stem cells and their use in engineering cell sheets to treat limbal stem cell deficient patients.


Assuntos
Ensaios Clínicos como Assunto , Células Epiteliais/transplante , Limbo da Córnea/patologia , Mucosa Bucal/citologia , Células-Tronco/patologia , Humanos , Engenharia Tecidual
15.
J Oral Pathol Med ; 49(4): 342-349, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31788854

RESUMO

BACKGROUND: Small extracellular vesicles (SEVs) have a diameter between 30 and 150 nm and play a key role in cell-cell communication. As cells cultured in 3D vs 2D behave differently, this project aimed to assess whether there were differences in SEVs derived from human oral mucosa lamina propria-progenitor cells (OMLP-PCs) cultured in a 3D matrix compared with traditional 2D monolayer cultures. METHODS: OMLP-PCs were cultured in 3D type I collagen matrices or on traditional 2D tissue culture plastic. Cell morphology and viability were assessed by light microscopy, actin staining, and trypan blue staining. SEVs secreted by OMLP-PCs were purified and quantitatively analyzed by a BCA assay and nanoparticle tracking analysis (NTA; nanosight™). SEVs were further characterized by flow cytometry. SEV proliferative function was assessed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: Cells cultured in 3D grew well as observed by light microscopy and phalloidin staining with cells branching in three dimensions (as opposed to the cells grown as monolayers on tissue culture plastic). NTA demonstrated a significantly higher number of SEV-sized particles in the conditioned medium of cells grown in 3D type I collagen matrices vs a 2D monolayer (P < .01). Like SEVs from 2D culture, SEVs from 3D culture demonstrated a particle size within the expected SEV range. Tetraspanin analysis confirmed that 3D-derived SEVs were positive for typical, expected tetraspanins. Cell proliferation analysis demonstrated that SEVs produced through 3D cell culture conditions significantly reduced the proliferation of skin fibroblasts when compared with SEVs from 2D monolayers (P < .05). CONCLUSION: 3D culture of OMLP-PCs produced typical SEVs but in a greater amount than when the same cells were cultured in 2D. The downstream proliferative potential of the SEVs was influenced by the initial culture methodology. Future work should now assess the potential effects of 3D SEVs on key wound healing activities.


Assuntos
Técnicas de Cultura de Células , Vesículas Extracelulares , Células-Tronco/citologia , Proliferação de Células , Células Cultivadas , Meios de Cultivo Condicionados , Humanos , Mucosa Bucal/citologia
16.
Eur J Pharmacol ; 867: 172837, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31811857

RESUMO

Cancer stem cells (CSCs) play a key role in carcinogenesis and progression of head and neck squamous cell carcinomas (HNSCC). The most common markers indicating for CSCs are: CD44, CD24, CD133, ALDH1A1. Our objective was to evaluate the prognostic potential of CSC markers in HNSCC. The study included 49 patients treated for primary HNSCC, 11 patients with upper respiratory tract epithelial dysplasia and 12 subjects with the normal pharyngeal mucosa as a control group. The frequency and expression levels of the four CSC markers were assessed by immunohistochemistry. Univariate and multivariate analyses were used to correlate CSC expression levels with tumor stage, lymph node metastases or overall survival (OS). CD44, CD24, CD133, ALDH1A1 were widely expressed in tumors, whereas CD44 was found to be higher in cancer tissue (P = 0.001). ALDH1A1 expression levels were found to be significantly higher in T3-T4 tumors vs. T1-T2 tumors (P = 0.05). Lymph node metastases had significantly higher expression levels of CD24 (P = 0.01) and CD133 (P < 0.05) than primary tumors. Multifactorial analysis revealed that overall survival (OS) for patients with ALDH1A1 negative tumors was 5.25 times higher than for patients with ALDH1A1 positive (ALDH1A1+) tumors (P = 0.01). On univariate and multivariate analysis, only ALDH1A1 positivity had a significant effect on OS of HNSCC patients (HR = 2.47 for P = 0.02). Immunohistochemistry-based assessments of CSC marker expression in HNSCC has significant predictive implications for patients with HNSCC. The frequency of CSCs in the tumor, specifically of ALDH1A1+ cells correlated with five-year OS in these patients.


Assuntos
/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias de Cabeça e Pescoço/mortalidade , Células-Tronco Neoplásicas/patologia , Retinal Desidrogenase/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/mortalidade , Idoso , Biomarcadores Tumorais/análise , Feminino , Seguimentos , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Hipofaringe/citologia , Hipofaringe/patologia , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Laringe/citologia , Laringe/patologia , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/citologia , Mucosa Bucal/patologia , Cultura Primária de Células , Prognóstico , Mucosa Respiratória/citologia , Mucosa Respiratória/patologia , Estudos Retrospectivos , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Taxa de Sobrevida , Células Tumorais Cultivadas
17.
Proc Natl Acad Sci U S A ; 117(38): 23329-23335, 2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-31611402

RESUMO

The development of biological markers of aging has primarily focused on adult samples. Epigenetic clocks are a promising tool for measuring biological age that show impressive accuracy across most tissues and age ranges. In adults, deviations from the DNA methylation (DNAm) age prediction are correlated with several age-related phenotypes, such as mortality and frailty. In children, however, fewer such associations have been made, possibly because DNAm changes are more dynamic in pediatric populations as compared to adults. To address this gap, we aimed to develop a highly accurate, noninvasive, biological measure of age specific to pediatric samples using buccal epithelial cell DNAm. We gathered 1,721 genome-wide DNAm profiles from 11 different cohorts of typically developing individuals aged 0 to 20 y old. Elastic net penalized regression was used to select 94 CpG sites from a training dataset (n = 1,032), with performance assessed in a separate test dataset (n = 689). DNAm at these 94 CpG sites was highly predictive of age in the test cohort (median absolute error = 0.35 y). The Pediatric-Buccal-Epigenetic (PedBE) clock was characterized in additional cohorts, showcasing the accuracy in longitudinal data, the performance in nonbuccal tissues and adult age ranges, and the association with obstetric outcomes. The PedBE tool for measuring biological age in children might help in understanding the environmental and contextual factors that shape the DNA methylome during child development, and how it, in turn, might relate to child health and disease.


Assuntos
Epigenômica/métodos , Células Epiteliais/metabolismo , Mucosa Bucal/citologia , Adolescente , Adulto , Criança , Pré-Escolar , Estudos de Coortes , Ilhas de CpG , Epigênese Genética , Feminino , Humanos , Lactente , Estudos Longitudinais , Masculino , Mucosa Bucal/metabolismo , Adulto Jovem
18.
J Oral Pathol Med ; 49(4): 320-327, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31145494

RESUMO

BACKGROUND: Candida albicans (C albicans) is the most common fungal pathogen causing opportunistic infections. IL17 (IL17A) is a vital mediator of antifungal immunity. The aim of the study was to investigate the effect of recombinant human interleukin 17A (rhIL17A) on human oral mucosal epithelial cells (hOMECs) defending against C albicans infection. METHODS: Human oral mucosal epithelial cells were divided into four groups: C albicans+ (MOI = 0.1), rhIL17A+ (100 µg/L), rhIL17A + C albicans+ (MOI = 0.1, rhIL17A:100 µg/L) and blank control. Then, C albicans growth was observed after 24 hours. Human beta-2 defensin (hBD-2), S100A8 and LL-37 in supernatants and their mRNAs in cells were measured by enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction, respectively. RESULTS: In C albicans+ group, C albicans hyphae formation and the death of infected hOMECs were observed. However, in the rhIL17A + C albicans+ group, IL17 inhibited both hypha formation, and C albicans from infecting hOMECs and its further growth. There was no statistical significance in adhesion rates of C albicans to hOMECs. Compared with the control group, the level of hBD-2 mRNA has increased, while hBD-2 and hBD-2 mRNA levels in the rhIL17A + C albicans+ group were the highest. Both hBD-2 and hBD-2 mRNA levels were higher in the rhIL17A+ group than in the C albicans+ group. S100A8 and LL-37 mRNAs have similar trend, and both upregulated after treatment with rhIL17A; however, protein levels were undetectable. CONCLUSION: Recombinant human interleukin 17A may inhibit C albicans from infecting hOMECs by affecting the growth and reproduction of C albicans as well as the formation of hyphae. Besides, rhIL17A might induce hBD-2, S100A8 and LL-37 secretion from hOMECs to strengthen their anti-infective ability.


Assuntos
Candida albicans/crescimento & desenvolvimento , Células Epiteliais/imunologia , Interleucina-17/farmacologia , beta-Defensinas/imunologia , Anti-Infecciosos , Peptídeos Catiônicos Antimicrobianos/imunologia , Calgranulina A/imunologia , Candida albicans/efeitos dos fármacos , Células Cultivadas , Células Epiteliais/microbiologia , Humanos , Mucosa Bucal/citologia , Proteínas Recombinantes/farmacologia
19.
Sci Rep ; 9(1): 18088, 2019 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-31792300

RESUMO

This study investigated a rabbit model of autologous simple oral mucosal epithelium transplantation (SOMET) for limbal stem cell deficiency (LSCD). LSCD was created in the SOMET group and the Control group. In the SOMET group, oral mucosa harvested from the buccal region was treated with dispase, cut into small pieces, and placed on the exposed corneal stroma without using graft sutures, amniotic membrane, and/or glue. A soft contact lens was positioned and tarsorrhaphy was performed in both groups. Postoperative corneal neovascularization and fluorescein staining scores were evaluated by slit lamp microscopy in both groups. At 2 weeks postoperatively, eyes were excised and subjected to immunohistochemical staining for CK3, CK13, CK15, and p63. In the SOMET group, transplantation of oral mucosa led to complete recovery of LSCD, as indicated by low neovascularization scores, low fluorescein staining scores, and detection of stratified K3/K13-positive cells on the stroma at 2 weeks after surgery. In contrast, corneal epithelial defects persisted in the Control group at 2 weeks. SOMET achieved re-epithelialization of the corneal surface in this rabbit LSCD model. It is a simple technique that does not require culture and could be a promising option for ocular surface reconstruction in bilateral LSCD.


Assuntos
Doenças da Córnea/terapia , Células Epiteliais/transplante , Mucosa Bucal/transplante , Animais , Doenças da Córnea/patologia , Células Epiteliais/citologia , Células Epiteliais/ultraestrutura , Feminino , Queratinas/análise , Limbo da Córnea/patologia , Mucosa Bucal/citologia , Mucosa Bucal/ultraestrutura , Coelhos , Técnicas de Cultura de Tecidos , Transplante Autólogo
20.
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
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...