Haagensenknowles4682
Skewing the macrophage polarity to achieve a favorable phenotype is a recently investigated therapeutic strategy in multiple disease/dysfunctional conditions such as inflammation, tumors, autoimmune disorders, and tissue repairs. However, delivering the therapeutic agent specifically to the macrophages has been a challenge in this field. Here, we describe the synthesis of hyaluronic acid (HA)-based nanoparticles for targeting CD44 receptors on the macrophages. The HA backbone is modified with cationic polyethyleneimine (PEI) for efficient encapsulation of microRNA into the self-assembling nanoparticles for targeted delivery to macrophages.Therapeutic gene delivery systems offer the potential for the treatment of a range of inherited and acquired inherited diseases. In contrast with viral gene vectors, the nonviral gene vectors provide a safer alternative and additional advantages such as the improved delivery efficiency, low cost, and often unlimited capacity to package DNA. Here we describe the preparation of a nonviral gene delivery technique based on lipid-peptide-DNA (LPD) complexes. The size of LPD particles is in the nanometer range. The use of these nanoparticulate LPDs results in high efficiency transfections and a high level of gene expression in vitro. LPDs provide a convenient and efficient tool for gene delivery in gene therapy.Crotamine is a basic, 42-residue polypeptide from snake venom that has been shown to possess cell-penetrating properties. Here we describe the preparation, purification, biochemical and biophysical analysis of venom-derived, recombinant, chemically synthesized, and fluorescent-labeled crotamine. We also describe the formation and characterization of crotamine-DNA and crotamine-RNA nanoparticles; and the delivery of these nanoparticles into cells and animals. Crotamine forms nanoparticles with a variety of DNA and RNA molecules, and crotamine-plasmid DNA nanoparticles are selectively delivered into actively proliferating cells in culture or in living organisms such as mice, Plasmodium, and worms. As such, these nanoparticles could form the basis for a nucleic acid drug-delivery system. We also describe here the design and characterization of crotamine-functionalized gold nanoparticles, and the delivery of these nanoparticles into cells. We also evaluated the viability of using the combination of crotamine with silica nanoparticles in animal models, aiming to provide slow delivery, and to decrease the crotamine doses needed for the biological effects. In addition, the efficacy of administering crotamine orally was also demonstrated.Biomimetic nanoparticles are hybrid nanostructures in which the uppermost layer is similar to a cell membrane. In these nanoparticles, lipids and biopolymers can be organized to improve drug incorporation and delivery. This report provides instructions for the preparation and physical characterization of four different biomimetic nanoparticles (1) polystyrene sulphate (PSS) nanoparticles covered with one cationic dioctadecyl dimethylammonium bromide bilayer (DODAB), which incorporates dimeric channels of the antimicrobial peptide Gramicidin D; (2) silica nanoparticles covered with one single bilayer of the antimicrobial cationic lipid DODAB; (3) hybrid lipid/polymer indomethacin (IND) nanoparticles from injection of IND/DODAB ethanolic solution in a water solution of carboxymethyl cellulose (CMC); (4) bactericidal and fungicidal nanoparticles from DODAB bilayer fragments (BF) covered consecutively by a CMC and a poly(diallyl dimethyl ammonium chloride) (PDDA) layer. These examples provide the basis for the preparation and characterization of novel biomimetic nanoparticles with lipids and/or biopolymers in their composition. The polymers and lipids in the hybrid nanoparticle composition may impart stability and/or bioactivity and/or provide adequate microenvironments for carrying bioactive drugs and biomolecules.In the development of drug delivery systems, researchers pursue multifunctionality to target more complex problems, while maintaining biocompatibility and high encapsulation efficiency. Herein, we describe the preparation of noncytotoxic particles with intrinsic antimicrobial properties able to entrap bioactive compounds. The particles are composed of a recombinantly produced elastin-like recombinamer functionalized with an antimicrobial peptide, and are spontaneously formed in mild conditions by exploiting the thermoresponsiveness of the elastin-like portion. This chapter provides advice and methods for the preparation of the self-assembled antimicrobial particles, the evaluation of antimicrobial activity and cytotoxicity, and the basis to set up the methodology for the encapsulation of bioactive compounds.Bacterial biofilms can cause problems in various arenas, from the fouling of water processing equipment to persistent in vivo infections. Silver nanoparticles are promising antimicrobial agents with activity against biofilm bacteria. Here we describe the synthesis of antimicrobial silver nanoparticles and the measurement of their antimicrobial activity against E. coli colony biofilms, which is a popular in vitro biofilm model for antibiotic assays.The increasing impact of metallic nanoparticles on life sciences has stimulated the development of new techniques and multiple improvements to the existing methods of manufacturing nanoparticles with tailored properties. Nanoparticles can be synthesized through a variety of physical and chemical methods. The choice of preparation procedure will depend on the physical and chemical characteristics required in the final product, such as size, dispersion, chemical miscibility, and optical properties, among others. Here we review basic practical procedures used for the preparation of protected and unprotected metallic nanoparticles and describe a number of experimental procedures based on colloidal chemistry methods. Y-27632 These include gold nanoparticle synthesis by reduction with trisodium citrate, ascorbic acid, or sugars in aqueous phase and nanoparticle passivation with alkanethiols, CTAB, or BSA. We also describe microwave-assisted synthesis, nanoparticle synthesis in ethylene glycol, and template-assisted synthesis with dendrimers, and, briefly, how to control nanoparticle shape (star-shaped and branched nanoparticles).