Anti-Inflammatory Role of Theobromine-Copper in NF-κB Pathway Inhibition in Wound Healing Models

Chronic wounds are characterized by persistent inflammation, matrix degradation, and impaired tissue regeneration. The NF-κB signaling pathway plays a central role in sustaining the inflammatory phase by promoting pro-inflammatory cytokine expression. Theobromine, a natural methylxanthine, has known anti-inflammatory properties but limited bioavailability. Copper, a trace element essential for wound repair, can enhance tissue regeneration and antioxidant responses. This study investigates the combined therapeutic effects of Theobromine-Copper nanoparticles (Theo-CuNPs) in modulating inflammation and promoting healing in periodontal ligament (PDL) cells. Methods: Theobromine-Copper nanoparticles were synthesized and characterized using UV–Vis spectroscopy, FTIR, and SEM. PDL cells were treated with Theo-CuNPs at 10, 50, and 100 µg/mL. Cell viability was assessed via MTT assay, and migration was evaluated using a scratch wound assay. Gene expression of nfkb1, il1b, tnfa, cox2, mmp9, and col1a1 was quantified using qRT-PCR after 24-hour treatment. Results: Theo-CuNPs demonstrated concentration-dependent effects. At 10 and 50 µg/mL, PDL cells retained normal morphology and viability, while 100 µg/mL showed cytotoxicity. Migration was enhanced at 10 µg/mL but reduced at higher concentrations. Gene expression analysis revealed significant downregulation of nfkb1, il1b, tnfa, cox2, and mmp9, and upregulation of col1a1 at 50 µg/mL, indicating an anti-inflammatory and regenerative response. Conclusion: Theobromine-Copper nanoparticles exert dual actions—suppressing inflammation via NF-κB pathway inhibition and promoting matrix regeneration through col1a1 upregulation. These findings suggest Theo-CuNPs as a promising biomaterial for inflammatory wound therapy, particularly in periodontal tissue repair.