RESUMEN
Low-level laser therapy has been investigated as a possible stimulus for enhancement of proliferation and differentiation of various cell types, but few reports relate undifferentiated mouse pulp cells (OD-21) response to irradiation in in vitro models. The aim of this study was to analyze the influence of low-level laser therapy (λ=660 nm), with three different irradiation times, on the behavior of OD-21 cell line. The cells were cultivated and divided into three groups: non-irradiated/control (group I); irradiated with 88 s (group II); irradiated with 177 s (group III) and irradiated with 265 s (group IV). Cell growth and viability were assessed after 7 and 10 days. Data were analyzed by Kruskal-Wallis and MannWhitney tests (α=.05). At day 7, there was a higher cell growth in groups I and II, as compared to group IV (p<.01). At the 10th day, group I showed a higher cell growth as compared to group II (p<.05). Cell viability in group IV was significantly lower at the 7th day, as compared to groups I (p<.001), II (p<.01) and III (p<.001). Cell viability in all the groups was over 80%, except in group IV at day 7. Irradiation time of group I influenced positively the proliferation and viability of OD-21 cells in late cell culture period. (AU)
A terapia a laser de baixa intensidade tem sido investigada como possível estímulo para aumento da proliferação e diferenciação de vários tipos de células, mas poucos relatos relacionam a resposta de células indiferenciadas da polpa dentária de camundongos (OD-21) à irradiação em modelos in vitro. O objetivo deste estudo foi analisar a influência do laser de baixa intensidade (λ=660 nm), com três períodos de irradiação diferentes, no comportamento das células da linhagem OD-21. As células foram cultivadas e distribuídas em três grupos: não irradiado / controle (grupo I); irradiado com 88 s (grupo II); irradiado com 177 s (grupo III) e irradiado com 265 s (grupo IV). O crescimento e a viabilidade celular foram avaliados após 7 e 10 dias. Os dados foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney (α = 0,05). No dia 7, houve crescimento celular maior nos grupos I e II, em comparação ao grupo IV (p <0,01). No décimo dia, o grupo I apresentou crescimento celular superior ao grupo II (p <0,05). A viabilidade celular no grupo IV foi significativamente menor no sétimo dia, em comparação aos grupos I (p <0,001), II (p <0,01) e III (p <0,001). A viabilidade celular em todos os grupos foi superior a 80%, exceto no grupo IV no dia 7. O tempo de irradiação do grupo I influenciou positivamente a proliferação e a viabilidade das células OD-21 no período mais tardio da cultura celular.A terapia a laser de baixa intensidade tem sido investigada como possível estímulo para aumento da proliferação e diferenciação de vários tipos de células, mas poucos relatos relacionam a resposta de células indiferenciadas da polpa dentária de camundongos (OD-21) à irradiação em modelos in vitro. O objetivo deste estudo foi analisar a influência do laser de baixa intensidade (λ=660 nm), com três períodos de irradiação diferentes, no comportamento das células da linhagem OD-21. As células foram cultivadas e distribuídas em três grupos: não irradiado / controle (grupo I); irradiado com 88 s (grupo II); irradiado com 177 s (grupo III) e irradiado com 265 s (grupo IV). O crescimento e a viabilidade celular foram avaliados após 7 e 10 dias. Os dados foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney (α = 0,05). No dia 7, houve crescimento celular maior nos grupos I e II, em comparação ao grupo IV (p <0,01). No décimo dia, o grupo I apresentou crescimento celular superior ao grupo II (p <0,05). A viabilidade celular no grupo IV foi significativamente menor no sétimo dia, em comparação aos grupos I (p <0,001), II (p <0,01) e III (p <0,001). A viabilidade celular em todos os grupos foi superior a 80%, exceto no grupo IV no dia 7. O tempo de irradiação do grupo I influenciou positivamente a proliferação e a viabilidade das células OD-21 no período mais tardio da cultura celular. (AU)
RESUMEN
ABSTRACT Sodium hypochlorite (NaOCl) remains the most used irrigation solution during root canal preparation because of characteristics such as wide-spectrum antimicrobial activity and organic tissue dissolution capacity. However, these solutions can alter dentin composition and there is no consensus on the optimal concentration of NaOCl to be used. Objectives To determine the organic matter dissolution and changes in dentin chemical composition promoted by different concentrations of NaOCl over time. Material and Methods: Fragments of bovine muscle tissue were weighed before and after 5, 10, and 15 min of immersion in the groups (n=10): G1- 0.9% saline solution; G2- 1% NaOCl; G3- 2.5% NaOCl; and G4- 5% NaOCl. Bovine dentin fragments were subjected to the same irrigants and absorption spectra were collected by Attenuated Total Reflectance of Fourier Transform Infrared Spectroscopy (ATR-FTIR) before and after 0,5, 1, 2, 3, 5, 8, and 10 min of immersion in the solutions. The ratios of the amide III/phosphate and carbonate/phosphate absorption bands were determined. The tissue dissolution and carbonate/phosphate ratios were submitted to the two-way analysis of variance (ANOVA) with Tukey’s multiple-comparison test (α<0.05) and to the one-way analysis of variance with Tukey’s (α<0.05). The amide III/phosphate ratio was analyzed by Friedman test (α<0.05) and the Kruskal-Wallis test with Dunn’s post-hoc (α<0.05). Results The increase in NaOCl concentration and contact time intensified the dissolution of organic matter and dentin collagen with reduction in the amide III/phosphate ratio. Significant differences between all groups (p<0.05) were observed in the dissolution of organic matter at 10 min and in the amide III/phosphate ratio between the saline solution and 5% NaOCl at 5 min. The carbonate/phosphate ratio decreased significantly in G2, G3, and G4 after 0,5 min of immersion (p<0.05), but more alterations did not occur in the subsequent periods (p>0.05). Intergroup differences were not observed in this ratio (p>0.05). Conclusions The increase in the exposure time and in the concentration of NaOCl solution lead to an increase in the tissue dissolution and dentin collagen deproteination. Furthermore, some carbonate ions are removed from the dentin inorganic phase by the NaOCl.
Asunto(s)
Animales , Bovinos , Hipoclorito de Sodio/química , Dentina/efectos de los fármacos , Dentina/química , Desinfectantes/química , Valores de Referencia , Irrigantes del Conducto Radicular/química , Solubilidad/efectos de los fármacos , Propiedades de Superficie/efectos de los fármacos , Factores de Tiempo , Colágeno/efectos de los fármacos , Espectroscopía Infrarroja por Transformada de Fourier , Concentración de Iones de Hidrógeno , Inmersión , Músculos/efectos de los fármacosRESUMEN
This work studied the immobilization of Cercospora kikuchii lipases on chitosan microparticles by chemical attachment on chitosan acetate microparticles activated by glutaraldehyde (CAM) added before or after the enzyme and physical adsorption on highly deacetylated chitosan hydrochloride microparticles (CHM). Lipases covalently immobilized on pre-activated CAM showed better performance retaining 88.4% of the enzymatic activity, with 68.2% of immobilization efficiency (IE). The immobilized enzyme retained an activity of about 53.5 % after five reuses, using p-NPP as substrate. Physical adsorption of lipase onto highly deacetylated CHM showed 46.2 % of enzymatic activity and 28.6% of IE. This immobilized derivative did not lose activity up to 80 days of storage at 4°C, while lipases immobilized on pre-activated CAM maintained its activity up to 180 days at same conditions. Taken together the results indicate that chitosan microparticles provide an optimal microenvironment for the immobilized enzyme to maintain good activity and stability.
RESUMEN
Estudos prévios têm demonstrado que o laser de Ho: YLF pode induzir mudanças fisicas e químicas na superfície do esmalte dentário, com o intuito de promover a prevençäo de cáries.Para tanto deve-se estabelecer uma dose limiar de irradiaçäo para que näo ocorram danos histológicos e/ou morfológicos aos tecidos moles e mineralizados. A temperatura na câmara pulpar em razäo da energia e freqüencia de irradiaçäo laser na superfície oclusal de dentes de coelhos foi avaliada "in vitro" neste estudo.(au)