Influence of titanium nanotubular surfaces, produced by anodization, on the behavior of osteogenic cells: in vitro evaluation / Influência da superfície nanotubular do titânio, produzido por anodização, no comportamento de células osteogênicas: avaliação in vitro

Objective: The objective of this study was to evaluate in vitro the influence of the anodized surface of Ti35Nb7Zr alloy on the behavior of osteogenic cells, for future application in biomedical implants. Material and Methods: For the development of this research, samples of commercially pure titanium (TiCp) and samples of Ti35Nb7Zr alloy were anodized, both were characterized by scanning electron microscopy (SEM) and were plated afterwards with human osteoblast-like cells (MG63 line) (2 x 104). Cell adhesion, cytotoxicity test, formation of mineralization nodules and a comet assay were also performed in different periods. The bottom of the plate was used as a control, without a sample. Results: SEM analysis showed that the topography of both samples presented surfaces covered by nanotubes. Cellular morphology exhibited spreading in both samples proposing an intimate cell- material liaison. After 3 days, the Ti35Nb7Zr group exhibited greater cell viability than the TiCp group (p<0.01). Regarding calcium content, there was no statistical difference between the anodized groups, but there was a difference between the experimental groups and the control group (p<0.01). In the comet assay, the percentage of DNA in the comet tail did not exhibit any significant difference (p>0.05) among the groups in the evaluated periods. Conclusion: It was concluded that this process of anodization was efficient to form nanotubes, as well as promote a positive influence on the behavior of osteogenic cells without promoting cell damage. (AU)

Objetivo: O objetivo deste estudo foi avaliar in vitro a influência da superfície anodizada da liga Ti35Nb7Zr no comportamento de células osteogênicas, para futura aplicação em implantes biomédicos. Material e Métodos: Para o desenvolvimento desta pesquisa, amostras de titânio comercialmente puro (TiCp) e amostras da liga Ti35Nb7Zr foram anodizadas, ambas foram caracterizadas por microscopia eletrônica de varredura (MEV) e posteriormente plaqueadas com células semelhantes a osteoblastos humanos (linha MG63) (2 x 104). Foram realizados em diferentes períodos a adesão celular, teste de citotoxicidade, formação de nódulos de mineralização e ensaio do cometa. O fundo da placa foi usado como controle, sem amostra. Resultados: A análise em MEV mostrou que a topografia de ambas as amostras apresentava superfícies cobertas por nanotubos. A morfologia celular exibiu espalhamento em ambas as amostras, propondo uma ligação íntima célula-material. Após 3 dias, o grupo Ti35Nb7Zr exibiu maior viabilidade celular do que o grupo TiCp (p<0.01). Em relação ao teor de cálcio, não houve diferença estatística entre os grupos anodizados, mas houve diferença entre os grupos experimentais e o grupo controle (p<0.01). No ensaio do cometa, a porcentagem de DNA na cauda do cometa não apresentou diferença significativa (p> 0.05) entre os grupos nos períodos avaliados. Conclusão:Concluiu-se que esse processo de anodização foi eficiente para formar nanotubos, além de promover uma influência positiva no comportamento das células osteogênicas sem promover dano celular. (AU)

In vitro evaluation of co-exposure of arsenium and an organic nanomaterial (fullerene, C60) in zebrafish hepatocytes.

Taking into account the concept of the "Trojan Horse", where contaminants may have its entry into specific organs potentiated by its association with nanomaterials, the aim of this study was to analyze the joint toxic effects induced by an organic nanomaterial, fullerene (C(60)) with the metalloid arsenic (As(III)). Hepatocytes of zebrafish Danio rerio were exposed to As(III) (2.5 or 100 µM), C(60) or As+C(60) for 4h, not altering cells viability. Intracellular reactive oxygen species concentration was reduced in cells exposed only to the C(60) (1mg/L) and in the treatment of 100 µM As(III)+C(60). Co-exposure with C(60) abolished the peak of the antioxidant glutathione (GSH) registered in cells exposed to the lowest As(III) concentration (2.5 µM). A similar result was observed in terms of lipid damage (TBARS). Total antioxidant capacity was significantly higher at both As(III) concentrations co-exposed to C(60) when compared with the control group. Activity of glutathione-S-transferase omega, a limiting enzyme in the methylation pathway of As(III), was reduced in the 100 µM As(III)+C(60) treatment. Cells co-exposed to C(60) had a significantly higher accumulation of As(III), showing a "Trojan Horse" effect which did not result in higher cell toxicity. Instead, co-exposure of As(III) with C(60) showed to reduce cellular injury.

Pollution biomarkers in estuarine animals: critical review and new perspectives.

In this review, recent developments in monitoring toxicological responses in estuarine animals are analyzed, considering the biomarker responses to different classes of pollutants. The estuarine environment imposes stressful conditions to the organisms that inhabit it, and this situation can alter their sensitivity to many pollutants. The specificity of some biomarkers like metallothionein tissue concentration is discussed in virtue of its dependence on salinity, which is highly variable in estuaries. Examples of cholinesterase activity measurements are also provided and criteria to select sensitive enzymes to detect pesticides and toxins are discussed. Regarding non-specific biomarkers, toxic responses in terms of antioxidant defenses and/or oxidative damage are also considered in this review, focusing on invertebrate species. In addition, the presence of an antioxidant gradient along the body of the estuarine polychaete Laeonereis acuta (Nereididae) and its relationship to different strategies, which deal with the generation of oxidative stress, is reviewed. Also, unusual antioxidant defenses against environmental pro-oxidants are discussed, including the mucus secreted by L. acuta. Disruption of osmoregulation by pollutants is of paramount importance in several estuarine species. In some cases such as in the estuarine crab Chasmagnathus granulatus, there is a trade off between bioavailability of toxicants (e.g. metals) and their interaction with key enzymes such as Na(+)-K(+)-ATPase and carbonic anhydrase. Thus, the metal effect on osmoregulation is also discussed in the present review. Finally, field case studies with fish species like the croaker Micropogonias furnieri (Scianidae) are used to illustrate the application of DNA damage and immunosuppressive responses as potential biomarkers of complex mixture of pollutants.