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1.
Braz. dent. sci ; 23(1): 1-8, 2020. tab, ilus
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-1049962

RESUMO

Objective: Dental composites developed by using nanotechnology in the field of dentistry are widely used in the treatment of anterior and posterior teeth. This study aimed to investigate the cytotoxic effects of dental composites of different particle size on L929 mouse fibroblast cell line by extract test method in vitro. Material and Methods: Composite samples of 8 x 2 mm diameter were prepared by polymerizing with led light device by using glass mod in a sterile cabinet. Composite samples of which surface areas were calculated according to ISO standards (3 cm2 / ml), were incubated for 24 and 72 hours, at 37 o C. cell viability was assessed by 3-[4,5-dimethylthiazole-2- yl]-2,5-diphenyltetrazolium bromide (MTT) assay and cell death was evaluated by the lactate dehydrogenase (LDH) leakage assay. Results: The 1:1 extracts of the composites at the end of 24 hours (except for nanoceramic composite) showed no toxic effect. When the cell viability results of the 1:1 extracts of the composite samples at the end of 72 hours were statistically analyzed, significant differences were found comparing to the control group (p < 0.05). Conclusion: It was observed that the type and size of the filler were effective on the toxicity of the composites, and the composites containing Bis-GMA, TEGDMA, UDMA and Bis EMA monomers in their organic matrix showed acceptable cell viability (70%) as specified by ISO. However, the composites with PEGDMA and BPA monomers in their organic matrix showed poor cell viability, which is below the acceptable level of 70%, and were found to have a toxic effect. (AU)


Objetivo: As resinas compostas desenvolvidas pela nanotecnologia no campo da odontologia são amplamente utilizadas no tratamento de dentes anteriores e posteriores. Este estudo teve como objetivo investigar os efeitos citotóxicos de resinas compostas de diferentes tamanhos de partículas na linha celular de fibroblastos de camundongos L929 pelo método de teste de extrato in vitro. Material e Métodos: Amostras compostas de 8 x 2 mm de diâmetro foram preparadas por polimerização com dispositivo de luz led usando um molde de vidro em um gabinete estéril. Amostras de resinas cujas áreas de superfície foram calculadas de acordo com os padrões ISO (3 cm2 / ml), foram incubadas por 24 e 72 horas, a 37 o C. A viabilidade celular foi avaliada pelo ensaio de brometo de 3- [4,5-dimetiltiazol-2- il] -2,5-difeniltetrazólio (MTT) e a morte celular foi avaliada pelo ensaio de infiltração de lactato desidrogenase (LDH). Resultados: Os extratos 1: 1 dos compósitos ao final de 24 horas (exceto o composto nanocerâmico) não apresentaram efeito tóxico. Quando os resultados de viabilidade celular dos extratos 1: 1 das amostras compostas ao final de 72 horas foram analisados, estatisticamente, foram encontradas diferenças significativas em relação ao grupo controle (p < 0,05). Conclusão: Observou-se que o tipo e tamanho da carga foram eficazes na toxicidade dos compósitos, e os compósitos contendo os monômeros Bis-GMA, TEGDMA, UDMA e Bis EMA em sua matriz orgânica apresentaram viabilidade celular aceitável (70%) como especificado pela ISO. No entanto, os compósitos com monômeros PEGDMA e BPA em sua matriz orgânica apresentaram baixa viabilidade celular, que está abaixo do nível aceitável de 70%, e foram encontrados como tendo um efeito tóxico. (AU)


Assuntos
Animais , Camundongos , Resinas Compostas/toxicidade , Estética Dentária , Fibroblastos , Técnicas In Vitro , Linhagem Celular , Sobrevivência Celular , Nanopartículas , L-Lactato Desidrogenase/toxicidade
2.
Int. j. morphol ; 37(4): 1229-1233, Dec. 2019. graf
Artigo em Inglês | LILACS | ID: biblio-1040117

RESUMO

SUMMARY: Cell culture is an important tool in medical, odontological and biological research laboratories, supporting cell therapies and tissue bioengineering strategies. Gingival fibroblasts present structural function, being able to modulate their metabolic capacity, which is reflected in the tissue morphology. The possibility of culturing fibroblasts in vitro, in monolayer or on three-dimensional scaffolds, for subsequent transplants in vivo opens important perspectives for the periodontal surgical clinic. The objective of the present article is to present a method of obtaining and cultivating viable human gingival fibroblasts for in vitro research. Explants derived from periodontal surgical discards were used, grown in 25 cm2 bottles to obtain a primary cell culture. After observing the proliferation and growth of the fibroblasts that interconnected and formed a monolayer network, involving the periphery of the explants, it was possible to remove the explants, to make the passage and the new subcultures were obtained in a ratio of 1:1. After 7 days, the amount of viable cells was analyzed in triplicate, using the Neubauer chamber technique, in cell culture bottles of 25 mm2 (T25) and 75 mm2 (T75). Fibroblasts were described and subclassified morphologically. The results showed a growth pattern in both bottles, but with a larger number in bottles of 75 cm2. Cells with fibroblastic morphology were subclassified into reticular and fusiform, being predominant those with fusiform morphology. In conclusion, culture of explant of human gingival connective tissue is a viable method for obtaining gingival connective tissue cells suitable for laboratory tests in cell culture, aiming at obtaining constructs for gingival tissue engineering.


RESUMEN: El cultivo celular es una herramienta importante en los laboratorios de investigación médica, odontológica y biológica, que apoyan las terapias celulares y las estrategias de bioingeniería de tejidos. Los fibroblastos gingivales presentan una función estructural, pudiendo modular su capacidad metabólica, que se refleja en la morfología tisular. La posibilidad de cultivar fibroblastos in vitro, en monocapa o en andamios tridimensionales, para trasplantes posteriores in vivo abre perspectivas importantes para la clínica de cirugía periodontal. El objetivo del presente artículo es presentar un método para obtener y cultivar fibroblastos gingivales humanos viables para investigación in vitro. Se utilizaron explantes derivados de los descartes quirúrgicos periodontales, crecidos en frascos de 25 cm2 para obtener un cultivo de células primarias. Después de observar la proliferación y el crecimiento de los fibroblastos que se interconectaron y formaron una red de monocapa, que involucraba la periferia de los explantes, fue posible eliminar los explantes, hacer el pasaje y los nuevos subcultivos se obtuvieron en una proporción de 1:1. Después de 7 días, la cantidad de células viables se analizó por triplicado, utilizando la técnica de cámara de Neubauer, en botellas de cultivo celular de 25 mm2 (T25) y 75 mm2 (T75). Los fibroblastos fueron descritos y sub-clasificados morfológicamente. Los resultados mostraron un patrón de crecimiento en ambas botellas, pero con un número mayor en botellas de 75 cm2. Las células con morfología fibroblástica se subclasificaron en reticulares y fusiformes, predominando aquellas con morfología fusiforme. En conclusión, el cultivo de explante de tejido conectivo gingival humano es un método viable para obtener células de tejido conectivo gingival adecuadas para pruebas de laboratorio en cultivos celulares, con el objetivo de obtener construcciones para la ingeniería del tejido gingival.


Assuntos
Humanos , Células do Tecido Conjuntivo , Técnicas de Cultura de Células/métodos , Bioengenharia/métodos , Gengiva/citologia , Biologia Celular , Fibroblastos
3.
Acta cir. bras ; 34(11): e201901101, Nov. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1054681

RESUMO

Abstract Purpose: To determine the efficacy of norbixin-based poly(hydroxybutyrate) (PHB) membranes for Achilles tendon repair. Methods: Thirty rats were submitted to total tenotomy surgery of the right Achilles tendon and divided into two groups (control and membrane; n = 15 each), which were further subdivided into three subgroups (days 7, 14, and 21; n = 5 each). Samples were analyzed histologically. Results: Histological analysis showed a significant reduction in inflammatory infiltrates on days 7, 14 (p < 0.0001 for both), and 21 (p = 0.0004) in the membrane group compared to that in the control group. There was also a significant decrease in the number of fibroblasts in the control group on days 7, 14 (p < 0.0001), and 21 (p = 0.0032). Further, an increase in type I collagen deposition was observed in the membrane group compared to that in the control group on days 7 (p = 0.0133) and 14 (p = 0.0107). Conclusion: Treatment with norbixin-based PHB membranes reduces the inflammatory response, increases fibroblast proliferation, and improves collagen production in the tendon repair region, especially between days 7 and 14.


Assuntos
Humanos , Animais , Masculino , Poliésteres/farmacologia , Tendão do Calcâneo/cirurgia , Tendão do Calcâneo/efeitos dos fármacos , Carotenoides/farmacologia , Tenotomia/métodos , Hidroxibutiratos/farmacologia , Valores de Referência , Regeneração/efeitos dos fármacos , Tendão do Calcâneo/patologia , Reprodutibilidade dos Testes , Ratos Wistar , Colágeno Tipo I/análise , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo III/análise , Colágeno Tipo III/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos
4.
Rev. bras. cir. plást ; 34(3): 391-398, jul.-sep. 2019. ilus
Artigo em Inglês, Português | LILACS | ID: biblio-1047162

RESUMO

Introdução: Queloides surgem de resposta excessiva à lesão da derme, resultando em proliferação de fibroblastos, produção exagerada de colágeno e comprometimento da pele sadia adjacente. O diagnóstico é clínico e muitos métodos conservadores e cirúrgicos já foram utilizados para tratamento. Porém, dados da eficácia desses tratamentos são limitados e não há consenso na literatura quanto a melhor técnica a ser empregada, permanecendo uma lacuna que necessita ser preenchida, a fim de que seus usos sejam indicados com maior confiabilidade, em um modelo de medicina baseada em evidências. Métodos: Revisão não sistemática da literatura sobre "queloides" nas bases de dados PubMed, Scielo, MEDLINE, UptoDate e livros-texto das áreas de Dermatologia e Cirurgia Dermatológica. Revisão de Literatura: Foram enumeradas e abordadas as principais informações sobre técnicas cirúrgicas e adjuvantes empregadas para essas lesões, que são: excisão, injeções intralesionais, crioterapia, laserterapia, revestimento com gel de silicone, radioterapia e pressoterapia. Torna-se relevante o levantamento dessas informações, tendo em vista que, além de poder causar dor, prurido e restrição de movimento, o principal motivo da procura de assistência médica para queloide é devido ao aspecto cosmético/estético, e as taxas de reincidência e falha terapêutica ainda são altas, sendo necessário conscientizar o paciente sobre o procedimento e seus efeitos. Conclusão: São muitos os tratamentos disponíveis para o queloide, sejam cirúrgicos ou não, todavia não há consenso sobre uma abordagem universalmente aceita. São necessários mais estudos, com a finalidade de definir a melhor conduta e atingir melhores resultados, visto a qualidade mediana das evidências apresentadas nos estudos.


Introduction: Keloids are characterized by an abnormal response to dermal trauma, resulting in fibroblast proliferation, excessive collagen production, and impairment of adjacent healthy tissue. The diagnosis is clinical, and many conservative and surgical methods can be used as treatments. However, data on the efficacy of these treatments are limited, and there is no consensus regarding the best treatment option. This gap needs to be filled by developing comprehensive evidence-based therapies. Methods: A non-systematic literature review of keloid scars was carried out using PubMed, Scielo, MEDLINE, UptoDate, and dermatology and dermatological surgery textbooks. Literature review: The search retrieved relevant information on surgical and adjuvant therapies used for keloids, including excision, intralesional injections, cryotherapy, laser therapy, silicone gel sheeting, radiation therapy, and pressure therapy. These data are crucial because, in addition to complaints of pain, itching, and restriction of movement, the main reason for seeking treatment for keloids is for cosmetic and aesthetic improvement, and the rates of recurrence and treatment failure are high, emphasizing the importance of creating awareness regarding the available procedures and their effectiveness. Conclusion: Many surgical and adjuvant therapies for keloids are available. Nonetheless, there is no consensus on a universally accepted treatment. Therefore, additional high-quality studies are needed to identify the most effective therapeutic approaches to achieve better results.


Assuntos
Humanos , História do Século XXI , Recidiva , Cirurgia Plástica , Terapêutica , Fator 1 de Crescimento de Fibroblastos , Fibroblastos , Procedimentos Cirúrgicos Dermatológicos , Queloide , Cirurgia Plástica/efeitos adversos , Cirurgia Plástica/métodos , Terapêutica/métodos , Ferimentos e Lesões , Ferimentos e Lesões/cirurgia , Ferimentos e Lesões/terapia , Fator 1 de Crescimento de Fibroblastos/análise , Fator 1 de Crescimento de Fibroblastos/efeitos adversos , Cicatriz , Cicatriz/complicações , Procedimentos Cirúrgicos Dermatológicos/métodos , Queloide/cirurgia
5.
An. bras. dermatol ; 94(4): 462-469, July-Aug. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1038307

RESUMO

Abstract: Cutaneous mucinoses are a heterogeneous group of dermatoses in which excess deposition of mucin in the dermis gives the skin a waxy appearance, with papules and plaques that can vary from self-healing mucinosis to even disrupting the normal shape of a patient's face, conferring a leonine facies, or be part of life threatening diseases like scleromyxedema. This review will describe the most recent classification on lichen myxedematosus in the generalized (scleromyxedema) and the localized forms, as well as the different organ systems involved in scleromyxedema, diagnostic workup, current management, and prognosis.


Assuntos
Humanos , Dermatopatias/diagnóstico , Dermatopatias/patologia , Escleromixedema/diagnóstico , Escleromixedema/patologia , Pele/patologia , Dermatopatias/classificação , Dermatopatias/terapia , Escleromixedema/classificação , Escleromixedema/terapia , Fibroblastos/patologia , Mucinas
6.
Rev. ADM ; 76(2): 72-76, mar.-abr. 2019. tab, graf
Artigo em Espanhol | LILACS | ID: biblio-1000403

RESUMO

Introducción: Los materiales para la obturación retrógrada son diversos. Actualmente, IRM y MTA son las alternativas clínicas más utilizadas, no obstante, es relativamente reciente la introducción de materiales a base de silicatos tricálcicos tal como Biodentine. Objetivo: Determinar la citotoxicidad de fibroblastos del ligamento periodontal humano expuestos a medios de cultivo condicionados con Biodentine, IRM y MTA. Material y métodos: 1 × 103 fibroblastos del ligamento periodontal humano fueron expuestos a medios DMEM/F12 condicionados con MTA, IRM y Biodentine en tres protocolos diferentes. Se realizó un ensayo de MTT para determinar la viabilidad celular a las cero, 24, 48, 72 horas, siete y 14 días. Se realizó una prueba ANOVA (p < 0.05). Resultados: En los tres protocolos con los diferentes medios de cultivo condicionados, la viabilidad de las células fue predominantemente proliferativa; sin embargo, las células expuestas a Biodentine mostraron una tendencia mayor que la MTA o la IRM. Conclusión: Las células expuestas a la Biodentine mostraron un comportamiento proliferativo a los 14 días de análisis. Se debe realizar más investigación a nivel in vivo y clínico para obtener más información sobre la conducta de estos materiales empleados para la obturación retrógrada (AU)


Introduction: The materials for retrograde filling are diverse. Currently, IRM and MTA are the most commonly used clinical alternatives, however, the introduction of materials based on tricalcium silicates such as Biodentine is relatively recent. Objective: To determine the cytotoxicity of human periodontal ligament fibroblasts exposed to culture media conditioned with Biodentine, IRM and MTA. Material and methods: 1 × 103 fibroblasts of the human periodontal ligament were exposed to DMEM/F12 media conditioned with MTA, IRM and Biodentine in 3 different protocols. An MTT assay was performed to determine cell viability at 0, 24, 48, 72 hours, seven and 14 days. An ANOVA test was performed (p < 0.05). Results: In the three protocols with the different conditioned culture media, the viability of the cells was predominantly proliferative, however, the cells exposed to Biodentine showed a higher tendency than the MTA or the IRM. Conclusion: The cells exposed to the Biodentine showed a proliferative behavior at 14 days of analysis. More research should be done at in vivo and clinical level to obtain more information about the behavior of these materials used for retrograde filling (AU)


Assuntos
Humanos , Materiais Restauradores do Canal Radicular/classificação , Materiais Restauradores do Canal Radicular/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Ligamento Periodontal , Obturação Retrógrada , Análise de Variância , Compostos de Cálcio , Compostos de Alumínio , Meios de Cultura , Fibroblastos
7.
Pesqui. bras. odontopediatria clín. integr ; 19(1): 4537, 01 Fevereiro 2019. graf, tab
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-998237

RESUMO

Objective: To analyze the effect of immediate placement of implants with extract from the new bone formation histometically. Material and Methods: In this true-experimental design with randomized post test control group, 9 mongrel dogs weighing 10 to 12 kg were used, which were divided into 3 groups, based on observation time of 14 days, 28 days and 56 days. On the installation of implants (∅3.5x10 mm) sequentially, the former socket extraction of the lower jaw's right second premolar tooth in the study sample injected 10% Aloe vera gel extract and left second left premolar tooth without injection of 10% Aloe vera extract. To compare independent groups use the Mann-Whitney test. All analysis were carried out using SPSS version 20. Results: There was an increase in the number of osteoblast cells in both treatment and control groups, but the value of the treatment group was greater. There were significant differences in the number of osteoblast cells between the treatment and control groups 14 days (p=0.019), 28 days: (p=0.018), and 56 days (p=0.009). There were no significant differences in the number of fibroblast cells between the treatment and control groups (p>0.05). But at observations 28 and 56 days, it was showed a significant difference in the number of fibroblast cells between the treatment and control groups (p=0.353 and p=0.024, respectively). Conclusion: Immediate placement of implants with 10% Aloe vera extract gel on extracted socket increases the number of osteoblasts and suppresses the number of osteoclasts and fibroblasts.


Assuntos
Animais , Cães , Osteoclastos , Células do Tecido Conjuntivo , Implantação Dentária Endo-Óssea , Aloe , Estatísticas não Paramétricas , Fibroblastos , Indonésia , Odontoblastos
8.
Acta cir. bras ; 34(2): e201900202, 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-989055

RESUMO

Abstract Purpose: To evaluate the hyaluronic acid (HA) inflammatory reaction, fibroblasts, fibrosis and duration of effect in the dorsal region of tobacco-exposed rats. Methods: Ten Wistar rats were divided into two groups: tobacco-exposed-group (TEG;n=5) and air-control-group (CG;n=5). The TEG animals were tobacco-exposed twice a day, 30-minutes/session, during 60 days. After this period, all animals received 0.1 mL HA subcutaneous injection in the dorsal area. The volume of HA was measured immediately after HA injection and weekly using a hand-caliper in nine weeks. After this period, all the animals were euthanized, and a specimen of was collected to evaluate inflammatory cells, fibroblasts, and fibrosis by HE. Results: This study showed a higher inflammatory reaction in TEG than CG: inflammatory cell-count (CG: 1.07±0.9; TEG: 8.61±0.36, p<0.001); fibroblast count (CG: 2.92±0.17; TEG: 19.14±0.62, p<0.001), and fibrosis quantification (CG: 2.0; TEG: 3.75, p<0.001). The analysis of the HA volume in nine weeks in the dorsal region did not show a difference between groups (p=0.39). Conclusions: This study suggested that the HA injection in the TEG caused an increase in inflammatory cell count, fibroblast, and fibrosis quantification when compared to the CG. There was no difference in the duration of effect of HA between the groups.


Assuntos
Animais , Masculino , Ratos , Tabaco/efeitos adversos , Exposição por Inalação/efeitos adversos , Viscossuplementos/efeitos adversos , Fibroblastos/efeitos dos fármacos , Ácido Hialurônico/efeitos adversos , Inflamação/patologia , Fatores de Tempo , Fibrose , Ratos Wistar , Modelos Animais de Doenças , Espaço Epidural/efeitos dos fármacos , Espaço Epidural/patologia , Fibroblastos/patologia , Inflamação/induzido quimicamente
9.
Araçatuba; s.n; 2019. 114 p. tab, graf, ilus.
Tese em Inglês, Português | LILACS, BBO - Odontologia | ID: biblio-1051133

RESUMO

Este trabalho foi dividido em dois capítulos que objetivou avaliar: 1) o efeito isolado ou combinado do flavonoide epigallocatechin-3-gallate (EGCG) em associação com o peptídeo LL-37 e seu análogo KR-12-a5 sobre a viabilidade celular de fibroblastos e sobre cultura planctônica, biofilme simples, dual-espécies e túbulos dentináios e 2) as interações sinérgicas do EGCG e proantocianidina do oxicoco (A-type cranberry proanthocyanidins, AC-PAC), quando usado em combinação com LL-37 ou KR-12-a5 sobre a viabilidade celular, a capacidade de migração e inibição das citocinas em cultura de fibroblastos (HGF-1), quando estimuladas ou não pelo lipopolissacarídeo de A. actinomycetencomitans (LPS). No capítulo 1, a concentração inibitória mínima (MIC), a concentração bactericida mínima (MBC) e concentração inibitória fracionária (FIC) de EGCG, LL-37 e KR-12-a5 foram determinadas a partir de valores decrescentes dos compostos por meio dos métodos de microdiluição e checkerboard contra Streptococcus mutans, Enterococcus faecalis, Actinomyces israelii e Fusobacterium nucleatum após 24 horas de tratamento. Fibroblastos da linhagem L-929 foram expostos a combinações de EGCG com peptídeos em diferentes concentrações e o metabolismo celular avaliado por ensaios de MTT. Os compostos com melhor efeito antimicrobiano e citotóxico foram avaliados por 24-36h, isoladamente ou em combinação, em biofilmes individuais ou biofilmes de dual-espécies com E. faecalis formados em placas de poliestireno por 48h por meio de contagem bacteriana. Os biofilmes de E. faecalis também foram cultivados em túbulos dentinários por 2 semanas, tratados com EGCG, KR-12-a5 e EGCG + KR-12-a5 e a porcentagem de células mortas foi determinada pela análise de imagens usando Microscopia Confocal. No capítulo 2, a linhagem celular de fibroblastos gengivais humanos primários HGF-1 foi pré-tratada durante 2 h com EGCG ou AC-PAC a 25 e 12,5 µg / mL, LL-37 ou KR-12-a5 a 0,06 e 0,03 µM ou com uma combinação de EGCG + ACPAC; AC-PAC + KR-12-a5; AC-PAC + LL-37; EGCG + KR-12-a5 ou EGCG + LL-37, nas mesmas concentrações. As culturas celulares foram então estimuladas com 50 µg/mL de LPS por 24-48h. A viabilidade celular e migração foram analisadas usando ensaios colorimétricos e fluorescentes, respectivamente. A quantificação de citocinas foi determinada por ensaios multiplex ELISA. Os resultados mostraram que em condições planctônicas, EGCG + KR-12-a5 apresentaram efeito sinérgico ou aditivo contra todas as bactérias testadas, com FIC menor que os valores de MIC obtidos pelos compostos isolados. As combinações de EGCG e peptídeos testados não foram tóxicas para os fibroblastos, uma vez que o crescimento celular foi superior a 70%. Em condições de biofilme simples, EGCG + KR-12-a5 eliminou S. mutans e A. israelii e reduziu E. faecalis e F. nucleatum. Para biofilmes de duas espécies, quando E. faecalis foi combinado com S. mutans, EGCG + KR-12-a5 teve efeito sinérgico eliminando S. mutans e reduzindo estatisticamente as contagens de E. faecalis. Em biofilmes associando E. faecalis e A. israelii ou F. nucleatum, EGCG + KR-12-a5 eliminaram E. faecalis e promoveram redução de A. israelii e F. nucleatum, embora não tenha sido observada diferença estatística entre os compostos. EGCG + KR-12-a5 reduziu mais de 80% dos biofilmes de E. faecalis nos túbulos dentinários. Dentre os grupos experimentais estudados, o EGCG, principalmente a 25 e 12,5 µg/mL estimulou o crescimento de fibroblastos, protegendo-os dos efeitos do LPS. Efeito sinérgico entre EGCG + AC-PAC, EGCG + LL-37 e EGCG + KR-12-a5 no metabolismo celular também foi observado na presença de LPS. Combinações do EGCG com AC-PAC ou KR-12-a5 e AC-PAC com LL-37 foram capazes de aumentar estatisticamente a migração celular. EGCG, AC-PAC, LL-37 e KR-12-a5 promoveram a redução de citocinas individualmente ou em combinação (EGCG + AC-PAC e EGCG + KR12-a5) mais especificamente para IL-6, IL-8, GM- CSF e TNF-α. Conclui-se que a associação de EGCG e KR-12-a5 é citocompatível e promove um efeito sinérgico contra bactérias associadas a infecções endodônticas, sob condições planctônicas e de biofilme. O EGCG, isoladamente ou associado ao AC-PAC e ao KR-12-a5, aumenta a viabilidade e migração celular, bem como a inibição de citocinas por fibroblastos estimulados por LPS. A associação de EGCG com KR-12-a5 poderia ser uma opção de princípio ativo em medicações para fins endodônticos(AU)


This study was divided in two chapters that aimed to evaluate: 1) the effect of flavonoid epigallocatechin-3-gallate (EGCG), cationic peptide LL-37 peptide and its analogue KR12-a5, alone or in combination, on fibroblast cell viability and on bacteria in planktonic and single/dual-species biofilms/dentin tubules; 2) the synergistic interactions of EGCG and cranberry proanthocyanidins (A-type cranberry proanthocyanidins, AC-PAC), when used in combination with LL-37 or KR-12-a5 on cell viability, the ability to induce cell migration and inhibit cytokines in culture of fibroblasts (HGF-1) when stimulated or not by the lipopolysaccharide of A. actinomycetencomitans (LPS). For the chapter 1, Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and fractional inhibitory concentration (FIC) of EGCG, LL-37 and KR-12-a5 were determined from decreasing values of the compounds by Streptococcus mutans, Enterococcus faecalis, Actinomyces israelii and Fusobacterium nucleatum against microdilution and checkerboard after 24 hours of treatment. L-929 fibroblasts were exposed to combinations of EGCG with peptides at different concentrations and cell metabolism assessed by MTT assays. The compounds if the best antimicrobial and cytotoxic effect were also evaluated for 24-36h, alone or in combination, in 48h singleor dual-species biofilms with E. faecalis formed on polystyrene plates by bacterial counting. E. faecalis biofilms were also cultured in dentin tubules for 2 weeks and treated with EGCG, KR-12-a5 and EGCG + KR-12-a5 to determine the percentage of dead cells by analysis of images using Confocal Microscopy. For the chaper 2, primary human gingival fibroblast HGF-1 cell line was pretreated for 2 h with either EGCG or AC-PAC at 25 and 12.5 µg/mL, LL-37 or KR-12-a5 at 0.03 and 0.06 µM or with a combination of EGCG + AC-PAC; AC-PAC + KR-12-a5; AC-PAC + LL-37; EGCG + KR-12-a5 or EGCG + LL37, at the same concentrations. Cell cultures were then stimulated with 50 µg/mL LPS for 24-48h. Cell viability and migration were analyzed using colorimetric and fluorescent assays, respectively. Quantification of cytokines was determined by multiplex ELISA assays. The results show that in planktonic conditions, EGCG + KR-12- a5 showed a synergistic or additive effect against all the bacteria tested, with FIC lower than the MIC values obtained by the compounds alone. Combinations of EGCG and peptides tested were not toxic to fibroblasts, since cell growth was higher than 70%. Under single biofilm conditions, EGCG + KR-12-a5 eliminated S. mutans and A. israelii and reduced E. faecalis and F. nucleatum. For dual- species biofilms, when E. faecalis was combined with S. mutans, EGCG + KR-12-a5 had a synergistic effect by eliminating S. mutans and statistically reducing E. faecalis counts. In biofilms associated with E. faecalis and A. israelii or F. nucleatum, EGCG + KR-12-a5 eliminated E. faecalis and promoted reduction of A. israelii and F. nucleatum, although no statistical difference was observed between the compounds. EGCG + KR-12-a5 reduced more than 80% of the E. faecalis biofilms in the dentin tubules. Among the experimental groups studied, EGCG, mainly at 25 and 12.5 µg/mL stimulated the growth of fibroblasts, protecting them from the effects of LPS. Synergistic effect between EGCG + AC-PAC, EGCG + LL-37 and EGCG + KR12-a5 on cell metabolism was also observed in the presence of LPS. Combinations of EGCG with AC-PAC or KR-12-a5 and AC-PAC with LL-37 were able to increase statistically cell migration. EGCG, AC-PAC, LL-37 and KR-12-α5 promoted cytokine reduction individually or in combination (EGCG + AC-PAC and EGCG + KR-12-a5) more specifically for IL-6, IL-8, GM-CSF and TNF-α. The association of EGCG and KR-12-a5 was cytocompatible and promoted a synergistic effect against bacteria associated with endodontic infections under planktonic and biofilm conditions. EGCG, alone or in combination with AC-PAC and KR-12-a5, increases cell viability and migration, as well as inhibition of cytokines by LPS-stimulated fibroblasts. The association of EGCG with KR12-a5 could be an option as active principle for medications to be used for endodontic purposes(AU)


Assuntos
Flavonoides , Biofilmes , Peptídeos Catiônicos Antimicrobianos , Citocinas , Fibroblastos
10.
Acta cir. bras ; 34(12): e201901202, 2019. graf
Artigo em Inglês | LILACS | ID: biblio-1054685

RESUMO

Abstract Purpose To explore the potential role and unclear molecular mechanisms of vaccarin in wound healing. Methods Rats' skin excision model to study the effects of vaccarin on wound healing in vivo . Hematoxylin and eosin staining was performed to evaluate Histopathologic characteristics. Immunohistochemistry was employed to assess the effects of vaccarin in accelerating angiogenesis. Western blot was used to evaluate relative protein expressed levels. Results Vaccarin could significantly promote wound healing and endothelial cells and fibroblasts proliferation in the wound site. Immunohistochemistry and Western blot studies showed that the nodal proteins and receptor (bFGFR) related to angiogenesis signaling pathway were activated, and the microvascular density in the wound site was markedly higher than that in the control group. Conclusions The present study was the first to demonstrate that vaccarin is able to induce angiogenesis and accelerate wound healing in vivo by increasing expressions of p-Akt, p-Erk and p-bFGFR. This process is mediated by MAPK/ERK and PI3K/AKT signaling pathways.


Assuntos
Animais , Masculino , Cicatrização/efeitos dos fármacos , Extratos Vegetais/farmacologia , Fosfatidilinositol 3-Quinases/efeitos dos fármacos , Quinases de Proteína Quinase Ativadas por Mitógeno/efeitos dos fármacos , Caryophyllaceae/química , Indutores da Angiogênese/farmacologia , Fatores de Tempo , Imuno-Histoquímica , Extratos Vegetais/química , Transdução de Sinais , Western Blotting , Reprodutibilidade dos Testes , Ratos Sprague-Dawley , Fosfatidilinositol 3-Quinases/análise , Quinases de Proteína Quinase Ativadas por Mitógeno/análise , Células Endoteliais/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/análise , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos
11.
Arch. argent. pediatr ; 116(6): 773-777, dic. 2018. ilus, graf
Artigo em Espanhol | LILACS, BINACIS | ID: biblio-973696

RESUMO

El síndrome de Sjogren-Larsson se caracteriza por retardo mental, ictiosis congènita y diplejía o cuadriplejía espástica. El defecto primario en este síndrome es la mutación del gen ALDH3A2, que codifica la enzima aldehído deshidrogenasa grasa y causa una deficiencia enzimática que produce una acumulación de alcoholes y aldehídos grasos en los tejidos que comprometen la integridad de la membrana celular, cuyos efectos pueden observarse en la piel, los ojos y el sistema nervioso central. El diagnóstico se realiza por medio de la cuantificación de la actividad de la enzima. Se describe el caso de una paciente con signos clínicos patognomónicos del síndrome de Sjogren-Larsson, cuyo diagnóstico se realizó por medio de la cuantificación de la actividad enzimática en un cultivo de fibroblastos. Además, tomando en cuenta el árbol genealógico de la paciente, se realizó el estudio en los padres y un hermano con signos sugestivos del síndrome de Sjogren-Larsson.


Sjogren-Larsson syndrome is characterized by congenital ichthyosis, mental retardation and spastic diplegia or quadriplegia. The primary defect in this syndrome is mutation of ALDH3A2 gen that codes for the fatty aldehyde dehydrogenase. Deficiency of this enzyme causes an accumulation of fatty alcohols and fatty aldehydes, leading to altered cell-membrane integrity. Skin, eyes, and the central nervous system are affected latter. The diagnosis is carried out through the cuantification of the enzyme activity.


Assuntos
Humanos , Feminino , Criança , Síndrome de Sjogren-Larsson/diagnóstico , Aldeído Oxirredutases/genética , Síndrome de Sjogren-Larsson/genética , Fibroblastos/enzimologia , Mutação
12.
Acta cir. bras ; 33(8): 703-712, Aug. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-949375

RESUMO

Abstract Purpose: To assess the action of vitamin C on the expression of 84 oxidative stress related-genes in cultured skin fibroblasts from burn patients. Methods: Skin samples were obtained from ten burn patients. Human primary fibroblasts were isolated and cultured to be distributed into 2 groups: TF (n = 10, fibroblasts treated with vitamin C) and UF (n = 10, untreated fibroblasts). Gene expression analysis using quantitative polymerase chain reaction array was performed for comparisons between groups. Results: The comparison revealed 10 upregulated genes as follows: arachidonate 12-lipoxygenase (ALOX12), 24-dehydrocholesterol reductase (DHCR24), dual oxidase 1 (DUOX1), glutathione peroxidase 2 (GPX2), glutathione peroxidase 5 (GPX5), microsomal glutathione S-transferase 3 (MGST3), peroxiredoxin 4 (PRDX4), phosphatidylinositol-3,4,5-trisphosphate dependent Rac exchange factor 1 (P-REX1), prostaglandin-endoperoxide synthase 1 (PTGS1), and ring finger protein 7 (RNF7). Conclusion: Cultured fibroblasts obtained from burn patients and treated with vitamin C resulted in 10 differentially expressed genes, all overexpressed, with DUOX1, GPX5, GPX2 and PTGS1 being of most interest.


Assuntos
Humanos , Masculino , Feminino , Adulto , Adulto Jovem , Ácido Ascórbico/farmacologia , Queimaduras/patologia , Expressão Gênica/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Valores de Referência , Pele/patologia , Araquidonato 12-Lipoxigenase/análise , Araquidonato 12-Lipoxigenase/efeitos dos fármacos , Queimaduras/tratamento farmacológico , Células Cultivadas , Estudos Transversais , Estatísticas não Paramétricas , Ubiquitina-Proteína Ligases/análise , Oxirredutases atuantes sobre Doadores de Grupo CH-CH/análise , Ciclo-Oxigenase 1/análise , Ciclo-Oxigenase 1/efeitos dos fármacos , Peroxirredoxinas/análise , Reação em Cadeia da Polimerase em Tempo Real , Oxidases Duais/análise , Oxidases Duais/efeitos dos fármacos , Glutationa Peroxidase/análise , Glutationa Peroxidase/efeitos dos fármacos
13.
An. bras. dermatol ; 93(2): 268-270, Mar.-Apr. 2018. graf
Artigo em Inglês | LILACS | ID: biblio-887174

RESUMO

Abstract: This study describes a case of a 19-year-old patient with seven asymptomatic lesions on the chest, measuring between 0.5 to 1cm in diameter, with no history of trauma in the region. The immunohistochemical evaluation was positive for vimentin and smooth muscle actin, determining Dermatomyofibroma as definitive diagnosis. Dermatomyofibroma is a benign skin tumor, with a myofibroblastic origin, prevalent in young women. It usually presents as a single lesion, with very few reports of multiple lesions.


Assuntos
Humanos , Feminino , Adulto Jovem , Neoplasias Cutâneas/patologia , Miofibroma/patologia , Biópsia , Imuno-Histoquímica , Cicatriz Hipertrófica/patologia , Fibroblastos/patologia
14.
Acta cir. bras ; 33(2): 144-155, Feb. 2018. graf
Artigo em Inglês | LILACS | ID: biblio-886256

RESUMO

Abstract Purpose: To evaluate the efficacy of the application of the human amniotic membrane (HAM) on the inflammatory process, fibroblast proliferation, formation of collagenand reduction of skin wound areas in rats. Methods: Thirty six rats were submitted to a surgical injury induction and divided into two groups (n = 18): group C (control) and T (treated with the HAM). The macroscopic evolution in the wound area and the histological characteristics of the skin samples were evaluated. Results: The regression of the wound area was greater in group T. The histological analysis revealed a significant reduction (p < 0.05) in the inflammatory infiltrate in group T at all experimental periods compared with that in the control group. Furthermore, the group T presented a significant increase in the proliferation of fibroblasts at 14 and 21 days compared with group C (p < 0.05). Regarding the deposition of mature collagen fibers, there was an increase in the replacement of type III collagen by type I collagen in group T (p < 0.05). Conclusion: Treatment with the HAM reduced the healing time as well as the inflammatory responses, increased the proliferation of fibroblasts, and induced a higher concentration of mature collagen fibers.


Assuntos
Humanos , Animais , Masculino , Ratos , Pele/lesões , Cicatrização/fisiologia , Curativos Biológicos , Colágeno/farmacologia , Âmnio/transplante , Pele/patologia , Cicatrização/efeitos dos fármacos , Distribuição Aleatória , Ratos Wistar , Colágeno Tipo I/metabolismo , Colágeno Tipo I/farmacologia , Colágeno Tipo III/metabolismo , Colágeno Tipo III/farmacologia , Modelos Animais de Doenças , Fibroblastos/metabolismo , Fibroblastos/patologia , Âmnio/química , Inflamação/metabolismo
15.
Pesqui. bras. odontopediatria clín. integr ; 18(1): 4085, 15/01/2018. tab, ilus
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-966895

RESUMO

Objective: To compare the potency of fibroblast cells proliferation in 12.5% and 25% Culture Media Conditioned Warton's Jelly (CMCWJ) and Advanced Platelet Rich Fibrin (A-PRF) cultured medium. Material and Methods: Fibroblast cells were divided into five groups: Group I (Control Group): serum-starved fibroblast without any treatment as a negative control; Group II: fibrolast that supplemented in 12.5% CMCWJ medium; Group III: fibrolast that supplemented 12.5% A-PRF medium; Group IV: fibrolast that supplemented 25% CMCWJ medium, and Group V: fibrolast that supplemented 25% A-PRF medium. The fibroblasts proliferation was counted by an automated cell counter. Statistical analysis was performed using One-way ANOVA and Post hoc Tamhane test was conducted to analyze the potential fibroblast proliferation differences in different concentration of CMCWJ and A-PRF group. Results: There were no significant differences in the fibroblast cell proliferation between GI and GIV, GII and GIV, GII and GIII, GII and GV, also GIV and GV. There were significant differences between GI and GII, GI and GIII, GI and GV, also GIII and GIV. Conclusion: The 12.5% CMCWJ group, 12.5% A-PRF group and 25% A-PRF group has excellent potential ability of fibroblast cells proliferation, meanwhile 25% CMCWJ group has the lowest mean potency of fibroblast cells proliferation compared to other groups. The 12.5% A-PRF Group has the highest mean of fibroblast cell proliferation amongst other groups.


Assuntos
Proliferação de Células , Geleia de Wharton/patologia , Fibroblastos/patologia , Fibrina Rica em Plaquetas , Indonésia
16.
Braz. oral res. (Online) ; 32: e17, 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889495

RESUMO

Abstract: Bulk-fill composites are claimed to be restorative materials used in deep preparations and effectively photoactivated in layers up to 4 mm. The aim of the present study was to evaluate the degree of conversion, post-gel volumetric shrinkage, and cytotoxicity of six bulk-fill and two conventional composites. Degree of conversion was determined by FTIR spectroscopy; post-gel volumetric shrinkage was determined using the strain gauge method; and cytotoxicity in human fibroblasts was evaluated indirectly by the MTT assay. Data were subjected to one-way ANOVA/Tukey's test (α = 0.05). All materials, including bulk-fill and conventional composites, were classified as non-toxic, with cell viability higher than 70%. Bulk-fill composites exhibited volumetric shrinkage similar to or lower (1.4 to 0.4%) than that of conventional composites (1.7-2.1%). However, only four of the bulk-fill composites were able to sustain a homogeneous conversion at the 4-mm depth. Despite their non-toxicity and shrinkage similar to that of conventional materials, not all commercial bulk-fill materials were able to maintain a conversion as high as 80% of the superficial layer, at the 4-mm depth, indicating some failure in the bulk-fill design of some commercial brands. Therefore, the use of bulk-fill materials in dental practice is advantageous, but special attention should be given to the selection and correct use of the materials.


Assuntos
Humanos , Resinas Compostas/toxicidade , Resinas Compostas/química , Polimerização , Valores de Referência , Propriedades de Superfície , Fatores de Tempo , Teste de Materiais , Sobrevivência Celular/efeitos dos fármacos , Reprodutibilidade dos Testes , Análise de Variância , Espectroscopia de Infravermelho com Transformada de Fourier , Transição de Fase , Processos Fotoquímicos , Fibroblastos/efeitos dos fármacos
17.
Braz. J. Pharm. Sci. (Online) ; 54(1): e00031, 2018. graf
Artigo em Inglês | LILACS | ID: biblio-889447

RESUMO

Preservatives are widely used substances that are commonly added to various cosmetic and pharmaceutical products to prevent or inhibit microbial growth. In this study, we compared the in vitro cytotoxicity of different types of currently used preservatives, including methylparaben, imidazolidinyl urea (IMU), and sodium benzoate, using the human newborn fibroblast cell line CCD1072Sk. Of the tested preservatives, only IMU induced a reduction in cell viability, as shown using the MTT assay and propidium iodide staining (IMU>methylparaben>sodium benzoate). IMU was shown to promote homeostatic alterations potentially related to the initiation of programed cell death, such as decreased mitochondrial membrane potential and caspase-3 activation, in the treated cells. Methylparaben and sodium benzoate were shown to have a very low cytotoxic activity. Taken together, our results suggest that IMU induces programed cell death in human fibroblasts by a canonical intrinsic pathway via mitochondrial perturbation and subsequent release of proapoptotic factors


Assuntos
Conservantes Farmacêuticos/análise , Aditivos em Cosméticos , Ciclo Celular , Citotoxicidade Imunológica , Potencial da Membrana Mitocondrial , Fibroblastos , Citometria de Fluxo/métodos
18.
Acta odontol. latinoam ; 31(1): 11-15, 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-909925

RESUMO

The aim of this study was to evaluate the cytotoxic effects of 2.5% sodium hypochlorite (NaOCl), 17% ethylenediamine tetraacetic acid (EDTA), and 1% peracetic acid (PAA) on human fibroblasts. FG11 and FG15 cell lines were cultured in 24well cell culture plates for cell proliferation assessment and 96well cell culture plates for the methylthiazolyldiphenyltetrazolium bromide (MTT) assay; Dulbecco's modified Eagle's medium (DMEM) was used as control data. The experimental solutions were used at 0.01%, 0.05%, and 0.1% dilutions and assessed at 1, 2, and 4hour intervals. Data were subjected to statistical analysis by twoway analysis of variance (ANOVA), followed by the Bonferroni test at a significance level of p <0.05. The assessment of cell proliferation in this study showed cytotoxicity to the fibroblasts with 2.5% NaOCl for all three dilutions at all time intervals, 17% EDTA for the 0.05% and 0.1% dilutions at the 2and 4hour intervals, and 1% PAA for all three dilutions at the 4hour interval. The cell viability assay (MTT assay) for fibroblasts showed 2.5% NaOCl to be cytotoxic at the 0.05% and 0.1% dilutions at all time intervals, 17% EDTA to be cytotoxic at the 0.1% dilution at the 2and 4hour intervals, and 1% PAA to be cytotoxic at the 0.1% dilution at the 2and 4hour intervals. In conclusion, 1% PAA was less cytotoxic than 2.5% NaOCl and 17% EDTA (AU)


O objetivo do presente estudo foi o de avaliar os efeitos citotóxicos de hipoclorito de sódio (NaOCl) a 2,5%, ácido etilenodiaminotetracético (EDTA) a 17% e ácido peracético (PAA) a 1% em fibroblastos humanos. As linhagens celulares FG11 e FG15 foram colonizadas em 24well cell plates para avaliação da proliferação celularr e em 96well cell plates para o ensaio de MTT; O médio modificado Dulbecco's Eagle's (DMEM) foi usado como controle. As soluções experimentais foram usadas com diluições de 0,01%, 0.05%, e 0,1% e avaliadas com 1, 2 e 4 horas de intervalo. Os dados foram submetidos à análise estatística pelo twoway ANOVA, seguido do teste de Bonferroni com nível de significância de p <0.05. A avaliação da proliferação celular neste estudo mostrou o NaOCL a 2,5% como citotóxico nas 3 diluições e 3 intervalos de tempo, o EDTA a 17% nas diluições de 0,05% e 0,1% nos intervalos de 2 e 4 horas, e o PAA a 1% em todas as diluições no intervalo de 4 horas. O teste de viabilidade cellular (MTT) mostrou o NaOCl a 2,5% citotóxico a 0,05% e 0,1% em todos os intervalos, o EDTA a 17% citotóxico nas diluições de 0,1% nos intervalos de 2 e 4 horas e o PAA a 1% citotóxico na diluição de 0,1% nos intervalos de 2 e 4 horas. Como conclusão o PAA a 1% mostrouse menos citotóxico que o NaOCl a 2,5% e o EDTA a 17% (AU)


Assuntos
Humanos , Ácido Peracético , Hipoclorito de Sódio , Ácido Edético , Citotoxinas , Fibroblastos , Fatores de Tempo , Sobrevivência Celular , Análise Estatística , Análise de Variância , Meios de Cultura
19.
Bauru; s.n; 2018. 95 p. ilus, graf, tab.
Tese em Inglês | LILACS, BBO - Odontologia | ID: biblio-884465

RESUMO

Current knowledge supports the application of TiF4 varnish to protect against tooth caries and erosion; however, it is indispensable to know its cytotoxic potential and the mechanism involved on it before applying in patients. Therefore, this study aimed to evaluate 1) The cytotoxic effect of titanium tetrafluoride (TiF4) varnish compared with sodium fluoride (NaF) varnish on murine fibroblast (NIH/3T3), varying the fluoride concentration and time of treatment and 2) The percentage of apoptosis and its mechanism (both mitochondrial mediated by the Bcl-2 family- and death receptorpathways) in human gingival fibroblasts (HGF) and murine fibroblasts (NIH/3T3) treated with TiF4 varnish compared to NaF varnish for 6 h. Step 1) NIH/3T3 were exposed to NaF or TiF4 varnishes containing 0.95, 1.95 or 2.45% F, for 6, 12 or 24 h. MTT viability (n=6) and Hoescht/PI stain assays (n=3) as well as the cells morphology (HE, only for 24 h, n=3) and stiffness (AFM, only for 2.45% F, 6 or 12 h) were analyzed. Both varnishes, at 1.90 and 2.45% F, reduced cells viability by similar extent (33-86% at 6 h, 35-93% at 12 h, and 87-98% at 24 h) compared to control, regardless of the type of fluoride. TiF4 and NaF (2.45% F) reduced cell stiffness to a similar extent, but only TiF4 differed from control. Step 2) HGF and NIH/3T3 were exposed to NaF or TiF4 (2.45% F) varnishes for 6 h. Cells were examined by the TUNEL method using fluorescence microscope. The caspases-3, -8 and -9 activities were assessed. The cDNA for cytocrome c, Bax, Bad, Bcl-2, VDAC-1 and Fas-L was amplified by quantitative PCR (qPCR). Bax, Bcl-2 and Fas-L were further detected by western blot. Both fluorides similarly increased the percentage of apoptosis, while they failed in activating caspases-3, -8 and -9 for both types of cells. Bax/Bcl-2 ratio, cytochrome C and VDAC-1 gene expressions were not altered by both fluoride treatments. However, NaF varnish increased the amplification of Fas-L gene for NIH/3T3 and HGF, while TiF4 varnish induced lower Bad/Bcl-2 ratio expression compared to control for NIH/3T3, but not for HGF. No effect of the fluorides was detected in the proteins analysis. TiF4 and NaF have similar cytotoxicity on NIH/3T3, which is dependent on the F concentration and the exposure time. Both fluorides, at the studied conditions, similarly induce a low percentage of apoptosis, with consequent modest activation of Bcl-2 and Fas-L-dependent signaling pathways.(AU)


Conhecimento atual suporta a aplicação de verniz de TiF4 para proteção contra cárie e erosão dentárias; entretanto, é indispensável conhecer o seu potencial citotóxico e o mecanismo envolvido antes de aplicá-lo em pacientes. Portanto, o objetivo deste estudo foi avaliar 1) o efeito citotóxico do verniz de tetrafluoreto de Titânio (TiF4) comparado ao fluoreto de sódio (NaF), em fibroblastos NIH/3T3, variando a concentração de fluoreto e o tempo de tratamento 2) a porcentagem de apoptose e seus mecanismos (ambos mitocondrial mediado pela família Bcl-2 e pelo receptor de morte celular) em fibroblastos gengivais humanos (FGH) e fibroblastos murinos (NIH/3T3) tratados com verniz de TiF4 comparado com verniz de NaF por 6 h. Etapa 1) NIH/3T3 foram expostos a vernizes de NaF e TiF4 contendo 0,95, 1,95 ou 2,45% F, por 6, 12 ou 24 h. Ensaios de viabilidade por MTT (n=6) e Hoechst 33342/iodeto de propídeo (n=3) bem como a morfologia (HE, apenas para 24 h, n = 3) e a rigidez celular (MFA, apenas para 2,45% F, 6 ou 12 h) foram realizados. Ambos os vernizes com 1,90 e 2,45% F reduziram a viabilidade das células de forma semelhante (33-86% em 6 h, 35-93% em 12 h e 87-98% em 24 h) em comparação com o controle, independentemente do tipo de fluoreto. TiF4 e NaF (2,45%) reduziram de forma similar a rigidez celular, mas somente TiF4 diferiu do controle no período de 6 h. Etapa 2) FGH e NIH/3T3 foram tratadas com verniz de NaF ou TiF4 por 6h. As células foram examinadas pelo método de TUNEL, usando microscopia de fluorescência. A atividade das caspases -3, -8 e -9 foram avaliadas. O cDNA para citocromo C, Bax, Bad, Bcl-2, VDAC-1 e Fas-L foi amplificado e quantificado por PCR em tempo real (qPCR). A expressão das proteínas Bax, Bcl-2 e Fas-L foi quantificada por western blot. Ambos os fluoretos aumentaram de forma semelhante a porcentagem de apoptose, enquanto falharam na ativação de caspases-3, -8 e -9 para ambos tipos celulares. A expressão gênica da relação Bax/Bcl-2, do citocromo C e do VDAC-1 não foram alteradas por ambos fluoretos. No entanto, o verniz NaF aumentou a amplificação do gene Fas-L para ambas as células, enquanto que o verniz TiF4 induziu menor expressão da razão Bad/Bcl-2 em comparação com o controle para NIH/3T3, mas não para FGH. Nenhum efeito foi detectado na análise de proteínas. TiF4 e NaF apresentam citotoxicidade similar em NIH/3T3, a qual é dependente da concentração de F e do tempo de exposição. Ambos os fluoretos, nas condições estudadas, induzem uma baixa porcentagem de apoptose, com consequente modesta ativação das vias de sinalização dependentes de Bcl-2 e Fas-L.(AU)


Assuntos
Humanos , Animais , Camundongos , Cariostáticos/farmacologia , Fibroblastos/efeitos dos fármacos , Fluoretos Tópicos/farmacologia , Titânio/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Sobrevivência Celular/efeitos dos fármacos , Gengiva/citologia , Marcação In Situ das Extremidades Cortadas , Microscopia de Fluorescência , Células NIH 3T3 , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Fatores de Tempo
20.
Braz. oral res. (Online) ; 32: e004, 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889501

RESUMO

Abstract Radiation combined injury, a life-threatening condition, has higher mortality than simple radiation injury. The aim of the present study was to analyze the efficiency of Aloe vera and silver nanoparticles in improving the healing of ulcerated oral mucosa after irradiation. Thirty male Albino mice were divided into five groups: control, radiation, Aloe vera (AV), silver nanoparticles (NS), and AV+NS. The mice were exposed to whole body 6Gy gamma-radiation. After one hour, 20% acetic acid was injected into the submucosal layer of the lower lip for ulcer induction. The animals received topical treatment with the assigned substances for 5 days. Lip specimens were subjected to hematoxylin and eosin and anti alpha-smooth muscle actin immunohistochemical staining. Results demonstrated occurance of ulcer three days post irradiation in all groups except in the AV+NS group where only epithelial detachment was developed. After seven days, data revealed persistent ulcer in radiation group, and almost normal epithelium in the AV+NS group. A significant reduction of epithelial thickness was detected in all groups at the third day as compared to control. At the seventh day, only the AV+NS group restored the epithelial thickness. Area percent of alpha-smooth muscle actin expression was significantly decreased in radiation group at the third day followed by significant increase at the seventh day. However, all treatment groups showed significant increase in alpha-smooth muscle actin at the third day, which decreased to normal level at the seventh day. Our study demonstrated the efficiency of Aloe vera and silver nanoparticles in enhancing ulcer healing after irradiation.


Assuntos
Animais , Masculino , Camundongos , Aloe/química , Raios gama/efeitos adversos , Nanopartículas Metálicas/uso terapêutico , Úlceras Orais/tratamento farmacológico , Úlceras Orais/etiologia , Lesões Experimentais por Radiação/tratamento farmacológico , Prata/uso terapêutico , Ácido Acético , Actinas/análise , Administração Tópica , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/efeitos da radiação , Fibroblastos/efeitos dos fármacos , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Úlceras Orais/patologia , Lesões Experimentais por Radiação/patologia , Valores de Referência , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do Tratamento , Cicatrização/efeitos dos fármacos
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