Optimization of the Conditions for Plasmid DNA Delivery and Transfection with Self-Assembled Hyaluronic Acid-Based Nanoparticles

Abstract

Polymeric systems have been extensively studied as polyelectrolyte complexes to enhance the cellular delivery and transfection efficiency of genetic materials, such as plasmid DNA (pDNA). Here, self-assembled nanoparticles were formulated by complexation of hyaluronic acid (HA)-conjugated poly­(ethylene glycol) (HA–PEG) and poly­(ethylenimine) (HA–PEI), respectively, with pDNA creating relatively small, stable, and multifunctional nanoparticle complex formulations with high transfection efficiency. This formulation strategy offers high gene expression efficiency and negligible cytotoxicity in HeLa and A549 human lung cancer cell lines. To develop the ideal formulation, in vitro transfection efficiency was studied for three different nanoparticle formulations (HA–PEI/HA–PEG, HA–PEI, and HA–PEG) with different concentrations. The combination of the three polymers (HA, PEG, and PEI) was significant for the formulation to achieve the maximum gene expression results. The nanoparticles were found to be stable for up to a week at 4 °C conditions. Overall, these HA-based nanoparticles showed promising aspects that can be utilized in the designing of gene delivery vectors for cancer therapy.

Local Immunomodulation Using an Adhesive Hydrogel Loaded with miRNA‐Laden Nanoparticles Promotes Wound Healing

“…To prepare HA NPs, we conjugated 20 kDa HA with PEI or PEG, and used both HA conjugates at 50:50 (w/w) ratio to form miR‐223* loaded NPs as described before . The addition of HA‐PEI increased the efficiency of encapsulation of miRNA (≈98%, as quantified by Ribogreen RNA assay) owing to the strong interaction between PEI and miRNA (Figure S3A, Supporting Information).…”

“…However, the miRNA condensed inside the HA NPs was protected from nucleases, as demonstrated by the absence of an miR‐223* band. Overall, the modification of the HA polymer backbone with cationic branched PEI helped decrease the overall negative charge of the HA biopolymer and promote nucleic acid encapsulation (HA–PEI/miRNA nanoparticles) via electrostatic interactions . The high counterion density of these nanoparticles is also known to promote endosomal escape following cellular internalization via the “proton‐sponge effect.” Furthermore, the PEG‐modified HA polymer backbones were able to self‐assemble with miRNA‐loaded HA–PEI, yielding multifunctional nanoparticle complexes that could help increase the residence time of these nanocomplexes in the circulation.…”

“…Therefore, we aimed to develop a GelMA‐based adhesive hydrogel for the local delivery of miR‐223 5p mimic (miR‐223*) encapsulated in hyaluronic acid (HA)‐based nanoparticles (NPs). Two types of HA conjugates (i.e., HA‐polyethyleneimine (HA‐PEI) and HA‐polyethylene glycol (HA‐PEG)) were used to encapsulate miR‐223* based on the electrostatic interaction between PEI and miRNA . In particular, we used HA‐PEG for the synthesis of the NPs to increase the residence time of these nanocomplexes in the circulation.…”

Biodegradable Polymers for Gene Delivery

“…The interaction of siRNA and HA by van der Waals forces has been exploited for gene silencing in a CD44-positive human osteocarcinoma cell line (MG63) and in human mesenchymal stromal cells [184]. Nanoparticle formulations prepared by the complexation of HA conjugated PEG (HA-PEG) and HA-PEI produced excellent results in gene transfection and gene expression with negligible cytotoxicity in HeLa and A549 human lung cancer cell lines [185]. HA was complexed with PEI and the complex used to deliver MMP13 gene in a mouse model of liver fibrosis, with excellent results [186].…”

Polysaccharide nanoparticles: from fabrication to applications

“…In the field of drug/gene delivery, the hydrophobic-hydrophilic core-shell structure plays a very significant role in enhancing the stability, water solubility and biocompatibility of NP structures. [154][155][156] The polymer core-polysaccharide shell structure will not be discussed in detail in this section as it is not directly related to polysaccharide NPs, more related information of how the core-shell structure with a polysaccharide shell is utilised can be found in the literature. 157,158…”