Modulation of Macrophage Functional Polarity towards Anti-Inflammatory Phenotype with Plasmid DNA Delivery in CD44 Targeting Hyaluronic Acid Nanoparticles

Abstract

The purpose of this study was to modulate macrophage polarity from the pro-inflammatory M1 to anti-inflammatory M2 phenotype using plasmid DNA (pDNA) expressing interleukin-4 (IL4) or interleukin-10 (IL10)-encapsulated in hyaluronic acid-poly(ethyleneimine) (HA-PEI) nanoparticles (NPs). The HA-PEI/pDNA NPs with spherical shape, average size of 186 nm were efficiently internalized by J774A.1 macrophages. Transfection of HA-PEI/pDNA-IL4 and HA-PEI/pDNA-IL10 NPs increased IL4 and IL10 gene expression in J774 macrophages which could re-program the macrophages from M1 to M2 phenotype as evidenced by a significant increase in the Arg/iNOS level, and upregulation of CD206 and CD163 compared to untreated macrophages. Following intraperitoneal (IP) injection to C57BL/6 mice, HA-PEI NPs effectively targeted peritoneal macrophages over-expressing CD44 receptor. In an in vivo model of stimulated peritoneal macrophages, IP administration of HA-PEI/pDNA-IL4 and HA-PEI/pDNA-IL10 to C57BL/6 mice significantly increased the Arg/iNOS ratio and CD163 expression in the cells. Furthermore, HA-PEI/pDNA-IL10 NPs significantly increased peritoneal and serum IL10 levels which effectively suppressed LPS-induced inflammation by reducing level of TNF-α and IL-1β in peritoneal macrophages and in the peritoneal fluid. The results demonstrated that pDNA-IL10-encapsulate HA-PEI NPs skewed macrophage functional polarity from M1 toward an anti-inflammatory M2 phenotype which may be a promising platform for the treatment of inflammatory diseases.

WITHDRAWN: Macrophage-targeting Fasudil treatment protects liver from the ischemia/reperfusion injury by promoting M2 macrophage polarization

“…Macrophages/monocytes are key players in the modulation of immunological responses and inflammation [40,41]. The M1 or M2 macrophage phenotype is important for promoting inflammation or healing, respectively [6].…”

The Current State of Nanoparticle-Induced Macrophage Polarization and Reprogramming Research

“…In vivo models of C57BL/6 mice intraperitoneally injected with LPS and IFN-γ presented notably down-regulated expression of iNOS (from 255-fold to about five-fold relative to the LPS-treated control) level and up-regulated expression of CD163 level following treatment with HA-PEI/pDNA NPs [46]. Further study suggested that targeting CD44-overexpressed peritoneal macrophages by HA and the capacity of PEI for encapsulating nucleic acids made HA-PEI easily internalized by the macrophage.…”

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

“…Moreover, previous reports have shown that HA can be easily cleared from the body through different excretion processes with a total turnover of about 5 g of HA per day in humans . HA‐based NPs also hold great potential for the specific targeting of inflammatory macrophages during sustained inflammation because of the specific interaction between HA and membrane receptors such as CD44, which are highly expressed on macrophages . Upon the formation of the hydrogels containing miR‐223* loaded HA NPs, we optimized the mechanical, physical, and adhesive properties of the hydrogels to ensure that they can adhere to the native skin and cover the wound during the healing process.…”