Nonendocytic delivery of lipoplex nanoparticles into. Factors determining the superior performance of lipiddna. Pdf liposome and protein based stealth nanoparticles. Two cationic lipids with c18 unsaturated chains, n1,n12diamidinon4,n9dioleoylspermine and n1,n12diamidinon4linoleoyln9oleoylspermine, were more efficient in terms of gfp expression reduction compared to the other cationic lipids with shorter c12 12. Selfassembled lipoplexes of short interfering rna sirna. Vaginal instillation of smallinterfering rna sirna using liposomes has led to silencing of endogenous genes in the genital tract and protection against challenge from infectious disease. Scaffoldmediated delivery for nonviral mrna vaccines. Nanoparticle, ultrafine unit with dimensions measured in nanometers. The different groups include fullerenes, metal nps. Lipoplexes are released from endosome by a flipflap mechanism. Nanoplexes involve the nucleic acid rnai being associated with the particle or encapsulated by it. Intravaginal gene silencing using biodegradable polymer.
Nanoparticles as nonviral gene delivery vectors chinmayee sarita katragadda, prasanta kumar choudhury and p. The data in figure 4a indicate that the peg coating provided a longer plasma halflife as compared to lactose, and the opposite was observed when calculating the half. Pdnasirna delivery abstract small interfering rna sirna has been widely used as potential therapeutic for treatment of various genetic disorders. Electrospray production of nanoparticles for drugnucleic. These viruslike nanoparticles are also referred to as nonviral vectors. Effect of lactose on uptake by specific leukocytes. From these results, the optimal formulation of peg and fapegmodi. View the article pdf and any associated supplements and figures for a period of 48 hours. Review article nanoparticles for brain drug delivery. Static micromixercoaxial electrospray synthesis of. These nanostructured complexes, called lipoplexes, have shown to be extremely useful vehicles in gene therapy. N12,3dioleoyloxy propyln,n,ntrimethylammonium methyl sulphate can form lipoplexes with negatively charged genes to form nanoparticles by electrostatic interaction, providing high in vitro transfection e ciency 3. Their treelike structure consists of three characteristic elements.
A shotgun proteomics approach was used to compare human plasma protein binding capability with cationic liposomes, dnacationic lipid complexes lipoplexes, and lipidpolycationdna lpd complexes. Introduction to nanoparticle characterization with afm 1 revision. Liposomes have been recognized as carriers for drug delivery. Lpd complexes easily release their dna payload, while lipoplexes remain largely intact and accumulate at the cell nucleus.
Factors determining the superior performance of lipiddnaprotammine nanoparticles over lipoplexes. Characterization of sirna lipoplex loading into macrophages. Smaller 50100 nm homogeneous lipid nanoparticles lnp, formed by mixing sirnas with. These four major types of nanoparticles are all nonionic lipids.
In this work quantum dot mediated forster resonance energy transfer qdfret was first used to study and compare the cellular uptake and the. This breakthrough opened the opportunity for other nonviral vectors, such as polymers. Imaging flow cytometry imagestream, millipore analysis confirmed that the sirna lipoplexes were found in intracellular vesicles figure1a. A read is counted each time someone views a publication summary such as the title, abstract, and list of authors, clicks on a figure, or views or downloads the fulltext. The company tested the process using titanium oxide and managed to make spheres measuring 1100 nanometers in diameter, as well as a nucleus covered with many. Request pdf the use of lactose as an alternative coating for nanoparticles nanoparticle mediated drug delivery has long utilized pegylation as a mechanism for reducing uptake by the. In traditional delivery methods such as lipoplexes or polyplexes complexed with naked mrna strands, protein expression was generally. Cholesterolrich lipidmediated nanoparticles boost of. Theranostic lipoplexes are an integrated nanotherapeutic system with diagnostic imaging capability and therapeutic functions. Although the data in figure 3 provide a snapshot of lipoplexes associated with the plasma and blood cell fractions, we conducted a more indepth characterization of formulations containing 5% lactosylceramide and peg ceramide. Nano highperformance liquid chromatography coupled with a highresolution ltq orbitrap xl mass spectrometer was used to characterize and compare their protein corona.
Genlantis were used to load scrambled, nonhomologous sirna labeled with cy5. They can be classified into different classes based on their properties, shapes or sizes. Encapsulation of cas9 and sgrna plasmids in dotapdopecholesterolcholpeg and dotapdopecholesteroldspepeg liposomes has happened via electrostatic interactions between the positively charged cationic lipid dotap and the negatively charged pdna. Gene delivery by lipoplexes and polyplexes sciencedirect. The topic that we would like to emphasize is the formulationassembly of lipidbased nanoparticles np with diameter under 100 nm for delivering nucleic acid in vivo. Solid lipid nanoparticles and lipoplex liposomepolycationdna complex, also. Lipoplexes and polyplexes represent the two major nanocarrier systems for nucleic acid delivery. The complexes formed using lipidbased carrier molecules are referred to as lipoplexes and those formed with polymeric complexes are referred to as polyplexes. Visualizing lipidformulated sirna release from endosomes. In the present study, both chitosan nanoparticles and cationic lipids have been used to transfect pdna expressing. Negative stain tem imaging and nanoparticle tracking analysis nta revealed condensed sirna lipoplexes with average sizes of 177.
The particle size distributions of nanoparticles, nanoplexes, liposomes and lipoplexes were measured by the cumulant method using a lightscattering photometer. However, because of their size, charge and toxicity, they are not suitable for in vivo use. It is generally accepted that lipoplexes size affects the endocytosis pathway and resulting intracellular trafficking in cells. The surface layer usually consists of a variety of molecules such as. Nanoparticles exist in the natural world and are also created as a result of human activities. Recent advances in chitosanbased carriers for gene delivery. Lipoplexes are liposome structures characterized by a bilayer lipid membrane. This article addresses the mucosal transport barrier by adopting a hydrophilic and neutral surface onto cationic lipiddna nanoparticles to reduce the obstacle of permeation through the mucus. Conventional methods, such as bulk mixing, are not able to achieve this goal. Np are different from cationic lipidnucleic acid complexes lipoplexes and are vesicles composed of lipids and. Request pdf production of polycaprolactone nanoparticles with hydrodynamic diameters below 100 nm cancer is a worldwide increasing burden and its therapy is often challenging and causes severe. Solid lipid nanoparticles and lipoplex liposomepolycationdna complex, also called lipid nanoparticles, are currently used to deliver drugs and genes to. Xray diffraction xrd studies have revealed that different structures exist. Microfluidics synthesis of gene silencing cubosomes.
The use of lactose as an alternative coating for nanoparticles. Nps are tiny materials having size ranges from 1 to 100 nm. Figure 1 schematic illustration of the cholrich lipidmediated nanoparticle carrying cas9sgrna plasmids. All of these elements play specific roles and have a great impact on dendrimers functionality. Selfassembled messenger rna nanoparticles mrnanps for. Previous studies examining their uptake and intracellular unpacking rely on organic fluorophores fraught with low signal intensity and photobleaching. Factors determining the superior performance of lipid dnaprotammine nanoparticles over lipoplexes. Cytosolic delivery is the major obstacle to sirna drug development. The following sections detail some of the major viruslike nanoparticle systems that have.
A applications for nanoparticles while nanoparticles are important in a diverse set of fields, they can generally be classified as one of two types. Dendrimers are one of the nanoparticles with very interesting properties and abilities. Nep, on the contrary, delivered lipoplexes directly into the cell cytoplasm and showed much higher fluorescence intensity than in conventional transfection and bep. Introduction to nanoparticle characterization with afm. This shows the feasibility for fast translation of dmg. Effect of cationic lipid type in pegylated liposomes on.
To demonstrate proof of transient horizontal transfer, commercially available cationic transfection lipoplexes genesilencer. It has been proposed that, after exposure to a biological milieu, the unit of interest in the cellnanomaterial interaction is not the bare nanoparticle, but the complex made of the nanoparticle and its hard corona of associated plasma proteins. To maintain sterility of the lipoplexes, dna, sonicated liposomes, and f5pegdspe solutions were. Nonionic lipids are safe, nontoxic and biocompatible. Different types of delivery systems that include liposome, solid lipid nanoparticles, nanostructured lipid carriers, lipidpolymer hybrid nanoparticles, lipoplexes, and. For lipoplexes, these factors include lipid composition, the types of lipids and the lipidodn ratios. Both systems display comparable toxicity in all workable charge ratios. Upon ethanol removal, dnalipid nanoparticles genospheres were formed. Pdf lipid nanoparticles for ocular gene delivery researchgate. Lipidbased colloidal particles have been extensively studied as systemic gene delivery carriers.
The use of nanotechnology in modern pharmacotherapy. Fluorescently labeled lipoplexes were injected intravenously into balbc mice, and blood was harvested after 1 h. Pot synthesis of pegylated lipoplexes to facilitate. Cancer cell targeting of lipid gene vectors by protein corona. The lipoplexes moved to the center of the cells and the fluorescence intensity increased. The widely used dotapdna lipoplex was employed as a reference. Lipoplexes are able to protect their genetic cargo from degradation, and deliver inside mammalian cells. Engineering nanoparticles for targeted delivery of. Physical properties of nanoparticles nanoparticles consist of three layers. Many factors affect the cellular uptake and the following unpacking of nanoparticles. Dopesirna lipoplexes can effectively reduce the excretion by kidneys and. Review nanoparticle vaccines adopting viruslike features.
Electrospray production of nanoparticles for drugnucl eic acid delivery our research group has been the first to explore electrospray as a means to produce solid lipid nanoparticles, lipoplexes and polyplexes for drugnucleic acid delivery. Plk1 sirna lipoplexes inhibited tumor growth of kb xenografts, as well as that of fapegmodi. Several liposomes and lipoplexes complexes of gene and cationic. Two types of structures were observed in plain lipoplexes radler et al. For that purpose, investigators have used liposomes, lipoplexes, albumin nanospheres, micelles, nano emulsions polymers, and nanoparticles with antibodies, among others. The utility of using a protamminedna complex coated with a lipid envelope made of cationic 1,2dioleoyl3trimethylammonium propane dotap for transfecting cho chinese hamster ovary cells, hek293 human embryonic kidney cells, nih 3t3 mouse embryonal cells, and a17 murine cancer cells cells was examined.
Lipoplexes are typically formed by direct mixing between cationic liposomes and dna solutions. Lipidbased nanoparticles for nucleic acid delivery. Furthermore, a slight deviation in the nanoparticles particle size can create a. Uptake and intracellular fate of multifunctional nanoparticles. Cationic lipids 1, used for in vitro transfection, form positively charged heterogeneous complexes with nucleic acids, called lipoplexes 2. Nucleic acid nanoparticles at a crossroads of vaccines and.
Pdf factors determining the superior performance of. Although lipoplexes and polyplexes exhibited similar intracellular fate, polyplexes had higher dissociation rate than lipoplexes. Collectively, these data explain why lpd nanoparticles often exhibit superior performances compared to lipoplexes in trasfecting cells and represent a promising class of nanocarriers for gene delivery. Review article nanoparticles for brain drug delivery massimomasserini. Assembly of nucleic acidlipid nanoparticles from aqueousorganic. Murthy royal college of pharmacy and health sciences, andhapasara road, berhampur, ganjam, orissa 760002 author for correspondence. Because of their size, they have unique material characteristics, and manufactured nanoparticles have practical applications in a variety of areas. Our results demonstrate the great potential of electrospray to produce nanoparticles in a wellcontrolled. Dope has the ability to catalyze the fusion process by undergoing from bilayer to inverted hexagonal structures 123. Pdf uptake and intracellular fate of multifunctional.
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