Xanthan gum-functionalised span nanoparticles for gene targeting to endothelial cells.

Endothelial cells play a critical role in many physiological processes; therefore, there is increasing evidence that the future of many treatments for pathologies depends on the development of endothelium-targeting systems. Thus, we have incorporated the natural polysaccharide xanthan gum (XG) into sorbitan monooleate nanoparticles to provide them with a hydrophilic and negatively charged surface shell with stabilising properties and an inherent ability to target endothelial cells. Enhanced Green Fluorescent Protein plasmid (pEGFP) was incorporated into the nanosystem, and the protection ability and stability of this system was confirmed. Nanoparticle cytotoxicity and transfection capacity were successfully tested in Human Umbilical Vein Endothelial Cells (HUVECs) before confirming their biocompatibility in vivo. Finally, biodistribution studies after pEGFP-XG nanoparticle systemic administration to mice evidenced GFP expression in the vascular endothelium of lung, liver and kidney, thus confirming the potential of xanthan gum-functionalised span nanoparticles for gene targeting to endothelial cells.

Development and characterisation of chondroitin sulfate- and hyaluronic acid-incorporated sorbitan ester nanoparticles as gene delivery systems.

Glycosaminoglycans (GAGs) are natural polymers that are broadly used in gene delivery systems to increase stability as well as decrease toxicity and nonspecific interactions, thereby increasing transfection efficiency. In this work, we propose sorbitan ester-based lipid nanoparticles (SENS) functionalised with the GAGs chondroitin sulfate (CS) and hyaluronic acid (HA) as gene delivery systems. For this purpose, we describe the design and evaluation of these nanosystems loaded with plasmid DNA, including an evaluation of their physicochemical characteristics, stability properties, ability to protect and efficiently transfect cells with Enhanced Green Fluorescent Protein plasmid (pEGFP) in vitro, and biocompatibility both in vitro and in vivo. We confirm that molecules with high biological value and targeting potential, such as HA and CS, can be successfully incorporated into our recently developed sorbitan ester-based nanoparticles (SENS) and that this incorporation leads to effective stabilisation of both nanosystems as well as protects plasmid DNA. We demonstrated that the aforementioned incorporation of HA and CS enables long-term stability of the nanosystems in both liquid and lyophilised states, which is a remarkable property that can aid in their transfer to industry. The ability of these functionalised nanosystems to transfect the A549 cell line without compromising cell viability was also shown, as well as their innocuous safety profile in vivo. Thus, we provide valuable evidence of the suitable properties and potential of these hybrid nanoparticles as gene delivery systems.

Nanocarriers for microRNA delivery in cancer medicine.

The number of deaths caused by cancer is expected to increase partly due to the lack of selectivity and undesirable systemic effects of current treatments. Advances in the understanding of microRNA (miRNA) functions and the ideal properties of nanosystems have brought increasing attention to the application of nanomedicine to cancer therapy. This review covers the different miRNA therapeutic strategies and delivery challenges for its application in cancer medicine. Current trends in inorganic, polymeric and lipid nanocarrier development for miRNA replacement or inhibition are summarized. To achieve clinical success, in-depth knowledge of the effects of the promotion or inhibition of specific miRNAs is required. To establish the dose and the length of treatment, it will be necessary to study the duration of gene silencing. Additionally, efforts should be made to develop specifically targeted delivery systems to cancer cells to reduce doses and unwanted effects. In the near future, the combination of miRNAs with other therapeutic approaches is likely to play an important role in addressing the heterogeneity of cancer.

Anionic nanoparticles based on Span 80 as low-cost, simple and efficient non-viral gene-transfection systems.

The existing strategies in the design of non-viral vectors for gene therapy are primarily conceived for cationic systems. However, the safety concerns associated with the use of positively charged systems for nucleic acid delivery and several reports regarding the efficacy of negatively charged systems highlights the need for improved gene-delivery vectors. With these premises in mind, we investigated the development of new negatively charged nanoparticles based on Sorbitan esters (Span(®)) ­ extremely cheap excipients broadly used in the pharmaceutical industry ­ on the basis of a simple, one-step and easily scalable procedure. For their specific use in gene therapy, we have incorporated oleylamine (OA) or poly-L-arginine (PA) into these nanosystems. Thus, we used Sorbitan monooleate (Span(®) 80) to design Span(®) 80-oleylamine and Span(®) 80-poly-L-arginine nanosystems (SP-OA and SP-PA, respectively). These systems can associate with the model plasmid pEGFP-C3 and show mean particle sizes of 304 nm and 247 nm and surface charges of -13 mV and -17 mV, respectively. The nanoparticles developed were evaluated in terms of in vitro cell viability and transfection ability. Both systems exhibited an appropriate cell-toxicity profile and are able to transfect the plasmid effectively. Specifically, the nanosystems including OA among their components provided higher transfection levels than the SP-PA nanoparticles. In conclusion, anionic nanoparticles based on Span(®) 80 can be considered low-cost, simple and efficient non-viral anionic gene-transfection systems.