Peritoneal Macrophage-Specific TNF-α Gene Silencing in LPS-Induced Acute Inflammation Model Using CD44 Targeting Hyaluronic Acid Nanoparticles

Abstract

The main goal of this study was to evaluate tumor necrosis factor-alpha (TNF-α) gene silencing in peritoneal macrophages upon activation with lipopolysaccharide (LPS), using CD44-targeting hyaluronic acid (HA)-based nanoparticles encapsulating TNF-α-specific small interfering RNA (siTNF-α). HA nanoparticles were formulated by blending hyaluronic acid-poly(ethylene imine) (HA-PEI), hyaluronic acid-hexyl fatty acid (HA-C6), and hyaluronic acid-poly(ethylene glycol) (HA-PEG) in 3:2:1 weight ratio, and encapsulating siTNF-α to form spherical particles of 78-90 nm diameter. Following intraperitoneal (IP) administration in LPS-treated C57BL/6 mice, the nanoparticles were actively taken up by macrophages and led to a significant downregulation of peritoneal TNF-α level. Downregulation of peritoneal macrophage-specific TNF-α also had a significant impact on other pro-inflammatory cytokine and chemokine levels in the serum. The C57BL/6 group of mice challenged with 5 mg/kg LPS had a significantly higher survival rate when they were treated with 3 mg/kg siTNF-α, either prior or simultaneously with the LPS administration, as compared to the LPS-challenged mice, which were treated with controls including the scrambled siRNA formulation. Overall, the results of this study demonstrate that CD44 targeting HA nanoparticles can selectively deliver siTNF-α to peritoneal macrophages leading to downregulation of pro-inflammatory cytokines in the peritoneal fluid and in the serum. This RNAi strategy could potentially provide an important therapeutic modality for acute inflammatory diseases, such as septic shock.

Natural Polysaccharides for siRNA Delivery: Nanocarriers Based on Chitosan, Hyaluronic Acid, and Their Derivatives

“…The presence of hyaluronic acid reduces the cytotoxicity of PEI-containing nanomaterials [136]. At the same time, branched PEI (bPEI) exercises a “proton sponge effect” in acidic intracellular compartments, which promotes the siRNA’s endosomal escape [137]. There are also a number of publications combining hyaluronic acid with chitosan for siRNA delivery, where chitosan improves the siRNA binding, while hyaluronic acid confers targeting ability and increases the stability of the delivery system (these literature examples and further details about the benefits of the association of these two natural polysaccharides can be found in the Section 5).…”

Hyaluronic acid-functionalized bilosomes for targeted delivery of tripterine to inflamed area with enhancive therapy on arthritis

“…These results indicate that HA@Tri-BLs are provided with superior targeted therapy for arthritis. We argue that this is associated with the high deformability of bilosomes and the specific affinity of HA@Tri-BLs to CD44 receptors largely expressed on inflamed tissues (Kosovrasti et al., 2016).…”

Polysaccharide nanoparticles: from fabrication to applications

“…Amiji and coworkers studied HA-based gene delivery. 237 HA was grafted with hexyl chains to introduce hydrophobicity, PEG to enhance particle stability and poly(ethylene imine) (PEI) to facilitate interaction with siRNA. Tumour necrosis factor-a-specific siRNA was encapsulated and the uptake as well as down regulation of tumour necrosis factor-a in macrophages observed.…”