I was wondering about one important aspect that warrants further clarification; specifically, the mechanistic distinction between the individual and synergistic effects of Cu2+ and luteolin versus their nanocomplexed form.
In the comparative in vivo experiments, particularly those involving DSS-induced colitis, CuL NCs demonstrated noticeably superior efficacy compared to Cu2+ alone, luteolin, or their physical mixture. Could the authors elaborate on how they ensured that these enhanced therapeutic effects stemmed specifically from the nanoformulation itself, rather than differences in pharmacokinetics, bioavailability, or cellular uptake between the free and complexed components? Were any targeted biodistribution analyses or release kinetics studies conducted to help differentiate these contributions?
Additionally, considering the redox-active nature of copper ions and the potential for cytotoxic effects, especially in inflamed environments where antioxidant defenses are already compromised, did the authors observe any signs of local or systemic oxidative stress in treatment groups receiving the higher dose (20 mg/kg) of CuL NCs?