Mannose-Functionalized Chitosan-TPGS/Tween 80 Nanocarriers for Macrophage Targeting: Enhanced Piperine Delivery to Potentiate Anti-Inflammatory and Antioxidant Therapy

Piperine (PIP), a plant alkaloid with anti-inflammatory and antioxidant effects, has poor solubility and bioavailability, limiting its therapeutic potential in macrophage-mediated inflammatory and oxidative stress conditions. Despite various nanocarrier systems being explored for bioactive compounds, the specific combination of mannose-functionalized chitosan with dual stabilizers (TPGS and Tween 80) for enhanced macrophage targeting and piperine delivery has not been investigated. We hypothesized that this novel formulation would significantly enhance piperine solubility, macrophage uptake, and anti-inflammatory/antioxidant effects compared to conventional systems, while modulating apoptosis-related pathways. This study evaluated targeted and non-targeted nanoparticles synthesized by ionic gelation and emulsification using RAW 264.7 and THP-1 macrophages. FTIR, UV–Vis, XRD, and CHNS confirmed mannose conjugation, while SEM, TEM, and AFM revealed morphology. Physicochemical properties were assessed by DLS, encapsulation efficiency (EE%), drug loading (DL%), and stability. Biological evaluations included drug release, cytotoxicity (MTT), apoptosis analysis (Annexin V–FITC/PI staining), cellular uptake (fluorescence microscopy with coumarin-6), anti-inflammatory assays (extracellular and intracellular NO inhibition, cytokine suppression), antioxidant activity (DPPH, ABTS, FRAP, TAC), intracellular ROS/RNS, and apoptosis-related markers. Targeted nanoparticles showed larger mean size (162 nm) versus non-targeted ones (78 nm). EE% was 82% (targeted) and 92% (non-targeted). Both demonstrated sustained 72 h release. Cellular uptake was significantly greater for targeted nanoparticles. Both formulations reduced NO and pro-inflammatory cytokines, regulated apoptosis-associated markers, and induced controlled apoptosis at higher concentrations, with stronger effects observed for targeted particles. Antioxidant activity increased dose-dependently, with targeted nanoparticles showing superior intracellular ROS/RNS suppression. This novel multi-functional platform efficiently encapsulates PIP, enhances macrophage targeting, modulates apoptosis pathways, and demonstrates superior therapeutic promise for inflammation-related disorders.