Andrea Masotti

Comparison of different commercially available cationic liposome-DNA lipoplexes: Parameters influencing toxicity and transfection efficiency.

Lipid-DNA complexes (lipoplexes) are widely used, since several years, as gene carriers. However, their transfection efficiency, both in vitro and in vivo, depends, in a rather complex way, on different interconnected parameters, ranging from the chemical composition of the lipid components to the size and size distribution of the complexes and, moreover, to the composition of the suspending medium. In this paper, we have investigated the behavior of nine different commercially available transfection agents (liposomal and non-liposomal) and their lipoplexes, at different molar charge ratios and in different experimental conditions. The size and the time stability of the resulting lipoplexes were investigated by means of dynamic light scattering methods and their toxicity and transfection efficiency were assayed in vitro in a model tumor cell line (C6 rat glioma cell line). An attempt to correlate the different parameters governing the complex phenomenology observed has been made. Whereas all the formulations investigated display a low toxicity, that increases with the increase of the lipid-DNA molar charge ratio, the transfection efficiency markedly depends, besides the molar charge ratio, on the lipid composition and on the lipoplex size, in a rather correlated way. The aim of this work is to present, in a wide scenario, an example of the inter-correlation among the different parameters that influence the transfection efficiency of lipoplexes and to suggest the role exerted by the average size of the resulting aggregates in their overall effectiveness as carriers in gene therapy.

A novel near-infrared indocyanine dye-polyethylenimine conjugate allows DNA delivery imaging in vivo

Near-infrared (NIR) fluorescence light has been applied to monitor several biological events in vivo since it penetrates tissues more efficiently than visible light. Dyes exhibiting NIR fluorescence and having large Stokes shift are key elements for this promising optical imaging technology. Here, we report the synthesis of a novel conjugate between a near-infrared indocyanine dye and an organic polyamine polymer (polyethylenimine, PEI) (IR820-PEI) with high chemical stability and good optical properties. IR820-PEI absorbs at 665 nm, emits at 780 nm, and displays a large Stokes shift (115 nm). Moreover, the reported conjugate is able to bind DNA, and the delivery process can be monitored in vivo with noninvasive optical imaging techniques. These characteristics make IR820-PEI one of the most effective and versatile indocyanine dye polymeric-conjugate reported so far.

Chitosan Micro- and Nanospheres: Fabrication and Applications for Drug and DNA Delivery

Polysaccharides and other cationic polymers have been recently used in pharmaceutical research and industry for their properties to control the release of antibiotics, DNA, proteins, peptide drugs or vaccines. They have been also extensively studied as non viral DNA carriers for gene delivery and therapy. Chitosan is one of the most used since it can promote long-term release of incorporated drugs. Here, we reviewed the recent literature on the preparation of chitosan micro- and nanospheres using different manufacturing processes (nanofabrication). Moreover, the preparation of chitosan and chitosan/DNA nanospheres using a novel and simple osmosis-based method has been recently reported. This novel nanofabrication method may be a useful alternative to obtain small DNA-containing nanospheres (38+/-4 nm) for biomedical applications. The reported method has general applicability to various synthetic or natural biopolymers. Solvent, temperature and membrane cut-off are the physicochemical parameters able to control the overall osmotic process leading to obtain several nanostructured systems with different size and shape that may be used in several biotechnological applications.

Common plants as alternative analytical tools to monitor heavy metals in soil.

BackgroundHerbaceous plants are common vegetal species generally exposed, for a limited period of time, to bioavailable environmental pollutants. Heavy metals contamination is the most common form of environmental pollution. Herbaceous plants have never been used as natural bioindicators of environmental pollution, in particular to monitor the amount of heavy metals in soil. In this study, we aimed at assessing the usefulness of using three herbaceous plants (Plantago major L., Taraxacum officinale L. and Urtica dioica L.) and one leguminous (Trifolium pratense L.) as alternative indicators to evaluate soil pollution by heavy metals.ResultsWe employed Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES) to assess the concentration of selected heavy metals (Cu, Zn, Mn, Pb, Cr and Pd) in soil and plants and we employed statistical analyses to describe the linear correlation between the accumulation of some heavy metals and selected vegetal species. We found that the leaves of Taraxacum officinale L. and Trifolium pratense L. can accumulate Cu in a linearly dependent manner with Urtica dioica L. representing the vegetal species accumulating the highest fraction of Pb.ConclusionsIn this study we demonstrated that common plants can be used as an alternative analytical tool for monitoring selected heavy metals in soil.

Polyethylenimine In Medicinal Chemistry

Polyethylenimine (PEI), an organic branched or linear polyamine polymer, has been successfully used in the past for DNA complexation and transfection in vitro and in vivo into several cell lines and tissues. PEI was also applied in different fields from gene therapy and several studies have emphasized the importance of this polymer in medicinal chemistry. In this brief critical review the uses and applications of this versatile polymeric molecule will be discussed.

Function/structure correlation in novel pH-dependent cationic liposomes for glioma cells transfection in vitro

Abstract Cationic liposomes are good candidates as gene carriers in cell biology due to their ability to bind DNA through electrostatic interactions. These liposomes are used as non-viral delivery systems in the gene therapy of glioma. pH-dependency and transfection efficiency of seven novel lipids ( MORF-1, MORF-2, MORF-3, PIPR-1, PIPR-2, MM54 and DC-Amy ) were studied. Two of these molecules ( PIPR-2 and MM54 ) show at specific charge ratios better transfection efficiency than that of some commercially available liposomes. To cite this article: C. Esposito et al., C. R. Chimie 6 (2003).