Targeted Co‐Delivery of Bioactives via Inhalable Folic Acid‐Functionalized Chitosan Microparticles Induces Immunomodulation and Selective Cytokine Production in A549 Lung Cancer Cells

ABSTRACT Despite advancements in cancer treatments, lung cancer is one of the most predominant cancer types responsible for a major contribution to cancer‐related mortality on a global scale. Folate receptor alpha (FRα) is overexpressed in many epithelial cancers, forming the basis of the “Trojan Horse” strategy in folate‐targeting micro/nanoparticles. Using the phytoblend of luteolin and rutin, this study involves the synthesis and optimization of folic acid‐functionalized bioactives‐loaded chitosan microparticles (FA_CM loaded ) as inhalables. Scanning electron microscopy (SEM) portrays distinctly separated microspheres, while the mass median aerodynamic diameter (MMAD) and hydrodynamic diameter reported ∼3.67 and ∼9.2 µm, respectively. The encapsulation efficiency, drug loading (%), drug release profile, and the Fourier transform infrared spectroscopy (FTIR) confirm the drug‐excipient compatibility in the microformulation. The immunomodulatory effect of the treatment protocol is established by the in vitro analysis of the activation of immune cell proliferation, cytokine quantification, and changes in biochemical parameters in cell lines L132 (derived from human embryonic lung tissue) and A549 (adenocarcinomic human alveolar basal epithelial cells). FA_CM loaded is reported to influence cellular functions without compromising cell viability on L132 and peripheral blood mononuclear cells (PBMCs) till 72 h, but selectively exhibits time‐dependent and dose‐dependent cytotoxic effect on A549 cells. This study encourages further in vivo validation to establish targeted delivery of flavonoids via the pulmonary route with increased bioavailability and minimal systemic toxicity.