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.
Highlights
High production of pro-inflammatory cytokines which contribute to inflammatory responses and tissue damage in inflammatory diseases[6]
The results indicated that J774A.1 macrophages were effectively re-polarized from M1 to M2 state by plasmid DNA (pDNA)-IL4 and pDNA-IL10 encapsulated in hyaluronic acid-poly(ethyleneimine) (HA-PEI) NPs
To confirm the macrophage polarizing effect by hyaluronic acid (HA)-PEI/pDNA-IL4 and HA-PEI/pDNA-IL10 NPs, we evaluated the change in the expression of CD206 and CD163 in J774A.1 macrophages treated with HA-PEI/pDNA-IL4 and HA-PEI/pDNA-IL10 NPs by FACS analysis
Summary
High production of pro-inflammatory cytokines which contribute to inflammatory responses and tissue damage in inflammatory diseases[6]. Targeting local peritoneal macrophages and modulating their phenotype may have potential for alleviating systemic inflammatory diseases. A gene therapy offers a novel approach for in vivo polarization of macrophages due to its ability to achieve long-term expression of nucleic acids replacing the frequent administration of the recombinant proteins[3]. There has been no report on targeted delivery system for repolarization of macrophages toward an anti-inflammatory M2 phenotype by gene therapy. We synthesized HA-PEI conjugate for encapsulation and specific delivery of plasmid DNA expressing IL4 and IL10 genes to macrophages for modulation of their functional polarity toward anti-inflammatory M2a and M2c phenotypes both in vitro in J774A.1 macrophages and in vivo in peritoneal macrophages of C57BL/6 mice
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