ABSTRACT
The use of nonviral carriers based on nanomaterials is a promising strategy for modern gene therapy aimed at protecting the genetic material against degradation and enabling its efficient cellular uptake. To improve the effectiveness of nanocarriers in vivo, they are often modified with poly(ethylene glycol) (PEG) to reduce their toxicity, limit nonspecific binding by proteins in the bloodstream, and extend blood half-life. Thus, the selection of an appropriate degree of surface PEGylation is crucial to preserve the interaction of nanoparticles with the genetic material and to ensure its efficient transport to the site of action. Our research focuses on the use of innovative gold nanoparticles (AuNPs) coated with cationic carbosilane dendrons as carriers of siRNA. In this study, using dynamic light scattering and zeta potential measurements, circular dichroism, and gel electrophoresis, we investigated dendronized AuNPs modified to varying degrees with PEG in terms of their interactions with siRNA and thrombin to select the most promising PEGylated carrier for further research.
Subject(s)
Metal Nanoparticles , Nanoparticles , Gold , Polyethylene Glycols , RNA, Small Interfering/genetics , ThrombinABSTRACT
Heterofunctionalized gold nanoparticles (AuNPs) were obtained in a one pot reaction of gold precursor with cationic carbosilane dendrons (first to third generations, 1-3G) and (polyethylene)glycol (PEG) ligands in the presence of a reducing agent. The final dendron/PEG proportion on AuNPs depends on the initial dendron/PEG ratio (3/1, 1/1, 1/3) and dendron generation. AuNPs were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), ultraviolet spectroscopy (UV-VIS), thermogravimetric analysis (TGA), nuclear magnetic resonance (1H NMR) and zeta potential (ZP). Several assays have been carried out to determine the relevance of PEG/dendron ratio and dendron generation in the biomedical properties of PEGylated AuNPs and the results have been compared with those obtained for non-PEGylated AuNPs. Finally, analyses of PEG recognition by anti-PEG antibodies were carried out. In general, haemolysis, platelet aggregation and toxicity were reduced after PEGylation of AuNPs, the effect being dependent on dendron generation and dendron/PEG ratio. Dendron generation determines the exposure of PEG ligand and the interaction of this ligand with AuNPs environment. On the other hand, increasing PEG proportion diminishes toxicity but also favors interaction with antibodies.
Subject(s)
Dendrimers/toxicity , Drug Carriers/toxicity , Gold/toxicity , Metal Nanoparticles/toxicity , Silanes/toxicity , Cations/chemistry , Cell Survival/drug effects , Chemistry Techniques, Synthetic/methods , Chemistry, Pharmaceutical/methods , Dendrimers/chemistry , Drug Carriers/chemistry , Dynamic Light Scattering , Erythrocytes/drug effects , Gold/chemistry , HeLa Cells , Humans , Leukocytes, Mononuclear , Magnetic Resonance Spectroscopy , Metal Nanoparticles/chemistry , Metal Nanoparticles/ultrastructure , Microscopy, Electron, Transmission , Platelet Aggregation/drug effects , Polyethylene Glycols/chemistry , Silanes/chemistry , Toxicity TestsABSTRACT
Cardiovascular diseases, including acute coronary syndromes (ACS), are one of the most serious problems of modern medicine and therefore every year 4 million Europeans have died. It is now believed that elevated levels of inflammatory factors in the blood promotes the development cardiovascular events and chronic inflammation plays a key role in the pathogenesis of atherosclerosis. Intensively conducted research in many centers in the world can confirm the desirability of introducing anti-inflammatory therapy to standard drug therapy. The balance between pro- and anti-inflammatory processes affect the risk of developing ACS.
Subject(s)
Acute Coronary Syndrome/pathology , Anti-Inflammatory Agents/therapeutic use , Atherosclerosis/pathology , Inflammation/drug therapy , Acute Coronary Syndrome/drug therapy , Acute Coronary Syndrome/prevention & control , Atherosclerosis/drug therapy , HumansABSTRACT
Atherogenesis is the process of atherosclerotic plaque formation, leading to coronary artery heart disease. This process involves immune cells, mainly T and B cells, monocytes and macrophages. The process of atherogenesis is induced by inflammatory damage of endothelial cells. The characteristic construction features of the atherosclerotic plaque is a predisposing factor for acute coronary syndromes. The accumulation of inflammatory cells in the artery inner membrane enhances the local inflammatory process due to the secretion of reactive oxygen species, inflammatory cytokines and metalloproteinases, which accelerate the development of atherosclerotic lesions in the arteries. In chronic inflammation of endothelial cells, which is atherosclerosis, there is a decrease in the concentration of elastin and collagen as a result of the increased apoptosis of smooth muscle cells of the intima. This reduces the integrity and strength of the fibrous cap that covers a layer of thrombogenic plaque from contact with blood elements. Permanent inflammation promotes the formation of necrotic core, composed of dead smooth muscle cells, macrophages and foam cells formed by phagocytosis of oxidized lipid molecules. The thin fibrous cap and a large necrotic core are the cause of plaque rupture and thrombus formation within the coronary artery.
