Alteration of Surface Glycoproteins After Photodynamic Therapy.

BACKGROUND: Cell membranes have been identified as an important intracellular cancer treatment target, since the glycoconjugates present on the cell surface are involved in numerous cell functions. Photodynamic therapy (PDT) is a therapeutic modality employed in the treatment of tumors that uses visible light to activate a photosensitizer. OBJECTIVE: This study analyzed the expression of surface carbohydrates after PDT with two different photosensitizers, 5-aminolevulinic acid (ALA) and Photosan-3. METHODS: Mice were injected subcutaneously with 2 × 105 B16 cells. After 7-10 days, the presence of a tumor with a diameter of 3.6 mm was observed. Photosan-3® and 5-aminolevulinic acid-ALA were used in the PDT treatment. Control animals (not submitted to either laser treatment or photosensitizer injection) and treated animals were euthanized 15 days post-treatment. The tumors were irradiated with a red diode laser, λ = 655 nm, energy density of 10 J.cm-2, and power density of 45 mW.cm-2. After 2 weeks of treatment with PDT, the mice were euthanized, the tumors were collected, and the cell surfaces were labeled with lectins concanavalin A (ConA) and wheat germ agglutinin (WGA). RESULTS: Fluorescence microscopy analysis of the cell surfaces with lectins ConA and WGA showed the presence of α-mannose and α-glucose. CONCLUSIONS: The combined effects of either Photosan-3 or ALA and red laser light on melanoma suggest an inhibitory glycosylation action from PDT on the surface of B16-F10 cells.

Analysis of indicators of osteogenesis, cytotoxicity and genotoxicity of an experimental ß-TCP compared to other bone substitutes / Análise dos indicadores da osteogênese, citotoxicidade e genotoxicidade de um ß-TCP experimental comparado a outros substitutos ósseos

The aim of this study was to evaluate the indicators of osteogenesis, cytotoxicity and genotoxicity of an experimental beta tri-calcium phosphate (experimental ß-TCP) compared with two other bone substitutes: bovine hydroxyapatite (HA) (Bio-Oss® - Geistlich) and beta tri-calcium phosphate (ß-TCP - Bionnovation). The cell viability and genotoxicity were measured by MTT and MNT assay, respectively. The indicators of osteogenesis were analyzed by alkaline phosphatase activity, total protein content, and calcium deposition. The MTT and MNT assay showed that none of the tested materials was cytotoxic nor genotoxic. Concerning the indicators of osteogenesis, it was observed that cells in contact with all the materials were able to induce the osteogenesis and this process was influenced by the period of the cell culture in contact with bone substitutes. Based on the results of this study, it was concluded that this experimental ß-TCP appears to be a promising material as a bone substitute.

O objetivo deste estudo foi avaliar os indicadores da osteogênese, citotoxicidade e genotoxicidade de um beta-tricálcio fosfato (ß-TCP experimental) comparado com dois outros substitutos ósseos : Hidroxiapatita Bovina (HA) (Bio-Oss® - Geistlich) e beta-tricálcio fosfato (ß-TCP - Bionnovation). A viabilidade celular e genotoxicidade foram mensuradas pelos ensaios MTT e MNT, respectivamente. Os indicadores da osteogênese foram analisados pela atividade de fosfatase alcalina (ALP), conteúdo de proteína total, e deposição de cálcio. Os ensaios MTT e MNT mostraram que nenhum dos materiais testados foi citotóxico ou genotóxico. Em relação aos indicadores da osteogênese, foi observado que as células em contato com todos os materiais foram capazes de induzir a osteogênese, e que esse processo foi influenciado pelo período da cultura celular em contato com os substitutos ósseos. Baseado nos resultados desse estudo, conclui-se que este ß-TCP experimental parece ser um material promissor para ser utilizado como substituto ósseo.

Subgingival Irrigation with a Solution of 20% Propolis Extract as an Adjunct to Non-Surgical Periodontal Treatment: A Preliminary Study.

Natural products, including propolis, are now frequently used to treat periodontal disease, but there are a few clinical studies in this area. The aim of this randomized clinical trial was to evaluate the effect of subgingival irrigation of periodontal pockets with a hydroalcoholic solution of propolis extract 20% (w/v) as an adjunct to periodontal therapy. Sixteen individuals were divided into a test group (TG), comprised 65 teeth (scaling and root planing + irrigation with propolis solution), and a control group (CG), comprised 62 teeth (scaling and root planing + irrigation with saline solution). Clinical data such as probing depth, plaque index, gingival index and oral hygiene index were collected at baseline (T0) and after 45 (T1), 75 (T2) and 90 (T3) days. Both groups showed significant differences among the evaluated periods. The TG presented more reduction (p < 0.05) of probing depth than CG at T1 and T3. Within the limits of this short-term study, these data suggest that irrigation with a hydroalcoholic solution of propolis extract 20% (w/v) as an adjunct in periodontal treatment was more effective than the mechanical treatment with saline solution in terms of reducing probing depth for up to 90 days from the beginning of treatment.

High doses of alcohol during pregnancy cause DNA damages in osteoblasts of newborns rats.

BACKGROUND: Alcohol exerts teratogenic effects and its consumption during pregnancy can cause deficit of bone development. The aim of the current study was to evaluate the genotoxic effects of prenatal exposure to ethanol on newborn rat osteoblasts. METHODS: Wistar rats were initially divided into two groups: Ethanol group which received Ethanol 20% V/V in liquid diet and solid diet ad libitum, and Control group, which received solid diet and water ad libitum. Each group received a specific diet for 8 weeks before breeding and throughout three weeks of gestation and the treatment was finished on the day the pups were killed. On the fifth day of life, the pups from each group were killed for removal of the calvaria and isolation of osteogenic cells by sequential enzymatic digestion. The cells were cultured for a maximum period of 14 days. The detection of genotoxic effects of alcohol was investigated by the comet and the micronucleus assay. RESULTS: Micronucleus and comet assay showed significant increases in DNA damage at 7 days in Ethanol group (p = 0.0302, p = 0.0446, respectively). However, at 14 days both assay showed no significant difference between the groups (p = 0.6194, p = 0.8326, respectively). CONCLUSION: Our results showed that prenatal exposure to ethanol induced DNA damage in osteoblasts, as shown by micronucleus formation and higher percentage of DNA in the comet tail. It can be concluded that prenatal exposure to ethanol damages osteoblast DNA in newborns exposed to high doses of ethanol during pregnancy, suggesting that prenatal ethanol consumption has a direct effect on fetal osteoblasts.

Titanium-35niobium alloy as a potential material for biomedical implants: In vitro study.

Research on new titanium alloys and different surface topographies aims to improve osseointegration. The objective of this study is to analyze the behavior of osteogenic cells cultivated on porous and dense samples of titanium-niobium alloys, and to compare them with the behavior of such type of cells on commercial pure titanium. Samples prepared using powder metallurgy were characterized using scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and metallographic and profilometer analyses. Osteogenic cells from newborn rat calvaria were plated over different groups: dense or porous samples composed of Ti or Ti-35niobium (Nb). Cell adhesion, cell proliferation, MTT assay, cell morphology, protein total content, alkaline phosphatase activity, and mineralization nodules were assessed. Results from XRD and EDS analysis confirmed the presence of Ti and Nb in the test alloy. Metallographic analysis revealed interconnected pores, with pore size ranging from 138 to 150µm. The profilometer analysis detected the greatest rugosity within the dense alloy samples. In vitro tests revealed similar biocompatibility between Ti-35Nb and Ti; furthermore, it was possible to verify that the association of porous surface topography and the Ti-35Nb alloy positively influenced mineralized matrix formation. We propose that the Ti-35Nb alloy with porous topography constitutes a biocompatible material with great potential for use in biomedical implants.

Efeito do álcool em osteoblastos de recém-nascidos de ratas submetidas ao consumo crônico de etanol / The effect of alcohol on rat newborns osteoblasts submitted to ethanol chronic consumption

O álcool atua no organismo podendo trazer várias doenças, entretanto sua ação no tecido ósseo ainda apresenta resultados controversos. O objetivo deste trabalho foi avaliar o efeito do consumo crônico de álcool a 20% em osteoblastos obtidos da calvária de ratos recém-nascidos. Foram utilizadas 18 ratas prenhas, tratadas durante a gestação e divididas em grupos conforme a dieta: álcool a 20%, grupo isocalórico, e controle. Aos três dias de vida, os recém-nascidos foram eutanasiados para remoção da calvária e isolamento das células por meio de digestão enzimática sequencial, sendo estas cultivadas por períodos de até 14 dias. Foram realizados testes para avaliar o efeito do álcool na adesão, proliferação e viabilidade celular, no conteúdo de proteína total, na atividade da fosfatase alcalina e nas formações nodulares de matriz mineralizada. Os resultados mostraram que em geral, a adesão celular não foi influenciada pelo consumo crônico de álcool, já que não foi demonstrada diferença estatística entre os grupos. Contudo, o grupo álcool apresentou aumento significativo na proliferação, exceto no período de 1 dia, e nas formações nodulares. Com relação a viabilidade celular, apenas no período de 3 dias houve aumento significativo de células no grupo álcool. Os valores representativos de proteína total variaram dependendo do período estudado, sendo maior no grupo controle com 7 dias, porém aos 14 dias houve maior média nos grupos isocalórico e álcool. Quanto à fosfatase alcalina observamos aumento de sua atividade nos grupos álcool e isocalórico em todos os períodos. Concluímos que nesta metodologia, o álcool não apresentou efeito deletério para os osteoblastos, talvez pelo curto tempo de administração

The alcohol affects the organism and may cause various diseases, eventhough its effects on bone metabolism are still controversial. The purposeof this paper was to evaluate the effects of alcohol 20% chronic consumption in osteoblasts obtained from the calvaria of newborn mice. The alcohol was administrated to pregnant mice throughout the entirepregnancy. For that purpose, 18 mice were used, divided in group saccording to the diet: 6 receiving alcohol 20%, 6 belonging to theisocaloric group and 6 receiving water and ration at will. At three days oflife, the newborns were euthanized so as to remove the calvaria and startthe cell culture procedures. The osteoblastic lineage cells were isolated by sequential enzymatic digestion and the osteoblasts were cultivated forperiods of 14 days or less. Tests were performed on the culture slides to evaluate the effect of alcohol based on adhesion, proliferation and cellular viability on total protein content, alkaline phosphatase activity and noduleformation of mineralized matrix. The results have shown that the alcoholgroup presented significant increase in proliferation, except for the periodof one day, and in nodule formation. A significant increase in the alcoholgroup concerning cellular viability was only observed in the period of 3 days and there was no statistic difference in adhesion. The total proteincontent was higher in the control group in 7 days, and the average higherin the isocaloric and alcohol groups in 14 days, according to the periods of time studied. An increase in the activity of the alkaline phosphatase was observed in the isocaloric and alcohol groups in all periods of evaluation. By this methodology, we have concluded that the alcohol has not presented any deleterious effect on osteoblasts, possibly due to the short period of administration