WebSketch

Actualmente o desenvolvimento de aplicações baseadas na Web é uma área em crescimento exponencial, graças aos benefícios que estas trouxeram consigo. Com a crescente utilização da Web e a sua evolução como plataforma, surgiram novas tecnologias que vieram revolucionar o desenvolvimento de aplicações sobre esta plataforma. Com interfaces mais ricas e consequentemente mais dinâmicas, este tipo de aplicações assemelham-se às típicas aplicações Desktop com a diferença que estão a ser executadas em um ambiente completamente distinto, um ambiente partilhado e de fácil acesso, sendo o browser a aplicação universal de acesso a qualquer aplicação Web. Designadas serviços, as aplicações Web fornecem funcionalidades semelhantes às das aplicações Desktop, sendo na maioria das vezes software gratuito. Sendo a Google a grande pioneira nesta área, outras grandes entidades viram aqui a oportunidade de distribuir o seu software de uma forma fácil e barata, ficando esta de imediato disponível a milhões de utilizadores. Embora as aplicações Web se assemelhem às aplicações Desktop, ao seu processo de desenvolvimento surgem um conjunto de novos desafios provocados pelo facto de estas se encontrarem em um ambiente completamente distinto.

PEGylated polyethyleneimine-entrapped gold nanoparticles for enhanced and targeted gene delivery applications

Gene therapy, which involves the transfer of nucleic acid into target cells in patients, has become one of the most important and widely explored strategies to treat a variety of diseases, such as cancer, infectious diseases and genetic disorders. Relative to viral vectors that have high immunogenicity, toxicity and oncogenicity, non-viral vectors have gained a lot of interest in recent years. This is largely due to their ability to mimic viral vector features including the capacity to overcome extra- and intra-cellular barriers and to enhance transfection efficiency. Polyethyleneimine (PEI) has been extensively investigated as a non-viral vector. This cationic polymer, which is able to compact nucleic acid through electrostatic interactions and to transport it across the negatively charged cell membranes, has been shown to effectively transfect nucleic acid into different cell lines. Moreover, entrapment of gold nanoparticles (Au NPs) into such an amine-terminated polymer template has been shown to significantly enhance gene transfection efficiency. In this work, a novel non-viral nucleic acid vector system for enhanced and targeted nucleic acid delivery applications was developed. The system was based on the functionalization of PEI with folic acid (FA; for targeted delivery to cancer cells overexpressing FA receptors on their surface) using polyethylene glycol (PEG) as a linker molecule. This was followed by the preparation of PEI-entrapped Au NPs (Au PENPs; for enhancement of transfection efficiency). In the synthesis process, the primary amines of PEI were first partially modified with fluorescein isothiocyanate (FI) using a molar ratio of 1:7. The formed PEI-FI conjugate was then further modified with either PEG or PEGylated FA using a molar ratio of 1:1. This process was finally followed by entrapment of Au NPs into the modified polymers. The resulting conjugates and Au PENPs were characterized by several techniques, namely Nuclear Magnetic Resonance, Dynamic Light Scattering and Ultraviolet-Visible Spectroscopy, to assess their physicochemical properties. In the cell biology studies, the synthesized conjugates and their respective Au PENPs were shown to be non-toxic towards A2780 human ovarian carcinoma cells. The role of these materials as gene delivery agents was lastly evaluated. In the gene delivery studies, the A2780 cells were successfully transfected with plasmid DNA using the different vector systems. However, FA-modification and Au NPs entrapment were not determinant factors for improved transfection efficiency. In the gene silencing studies, on the other hand, the Au PENPs were shown to effectively deliver small interfering RNA, thereby reducing the expression of the B-cell lymphoma 2 protein. Based on these results, we can say that the systems synthesized in this work show potential for enhanced and targeted gene therapy applications.