Ti₃C₂O₂ MXene as a dual-action modulator of inflammatory and tuberculosis signaling: structural and in vitro insights

The growing threat of antimicrobial resistance and tuberculosis (TB) necessitates innovative therapeutics capable of modulating both infection and host immune responses. In this study, we report the dual anti-tubercular and anti-inflammatory activity of oxygen-functionalized Ti₃C₂O₂ MXene, a two-dimensional nanomaterial synthesized via selective HF etching of Ti₃AlC₂ followed by ethanol-assisted delamination. Structural characterization by XRD and Raman spectroscopy confirmed successful conversion to the MXene phase, with an increased interlayer spacing of 9.87 Å. SEM and TEM analyses revealed uniform sheet-like morphology, and AFM confirmed few-layer thickness (∼1.5 nm). Biological assays using lipopolysaccharide (LPS)-stimulated murine macrophage cell line RAW 264.7 macrophages demonstrated significant downregulation of pro-inflammatory mediators- inducible nitric oxide synthase (iNOS), Tumor necrosis factor (TNF)-α, and Interleukin-6 (IL-6)-with IC₅₀ values of 23.2, 26.1, and 21.6 μg mL^-1, respectively. Western blot analysis further validated suppression of inflammatory pathways at the protein level. In parallel, Ti₃C₂O₂ exhibited robust anti-tubercular activity against Mycobacterium tuberculosis H37Rv, achieving complete inhibition at 4.0 μg mL^-1. Computational studies revealed strong and specific interaction of Ti₃C₂O₂ with the TB inflammatory target protein (PDB ID: 5V3Y), forming stable hydrogen bonds with His185, Gln186, and Asp219. Molecular dynamics simulations over 3000 ns confirmed a highly stable protein-MXene complex (RMSD: 2.41 Å; ΔG_bind: -78.54 kcal mol^-1). Comparative simulations with streptomycin revealed weaker binding and greater structural fluctuation. ADMET predictions suggested favorable pharmacokinetic properties, including high volume of distribution, low toxicity, and absence of major cytochrome P450 or cardiotoxic liabilities. These findings establish Ti₃C₂O₂ MXene as a promising nanoplatform for dual-function immunomodulation and antimicrobial therapy, offering mechanistic and structural insight into its bioactivity.