Evaluation of Antioxidant and Anti-Inflammatory Effects of a Nanoformulation Derived from Annurca Apple Callus Extract in an In Vitro Model of Iron Overload-Induced Inflammation

Ferroptosis, a regulated form of cell death driven by iron accumulation and lipid peroxidation, contributes to oxidative stress-related skin damage. This study evaluates the antioxidant and anti-inflammatory effects of a nanoformulation derived from an Annurca apple callus extract in an in vitro model of ferroptosis using human keratinocytes (HaCaT cells). A hydroalcoholic extract from light Annurca apple callus (LCE) was nanoformulated with Pluronic^® F127 and Soluplus^® to enhance stability and bioavailability. The resulting nanoformulation (NF-LCE) exhibited optimal particle size (103.17 ± 0.87 nm), polydispersity index (0.21 ± 0.00), and zeta potential (-1.88 ± 0.64 mV). Iron overload (100 µM) was employed to induce oxidative stress and inflammation in HaCaT cells, resulting in elevated levels of inflammatory markers (COX2, IL-6, TNF-α) and a diminished antioxidant response, as indicated by decreased expression of GPX4 and Nrf2. NF-LCE treatment restored GPX4 and Nrf2 levels (~0.8-fold increase, <i>p</i> < 0.05) while significantly reducing COX2 (36.6%, <i>p</i> < 0.01), IL-6 (79.6%, <i>p</i> < 0.0001), and TNF-α (30.9%, <i>p</i> < 0.1). These results suggest NF-LCE as a promising therapeutic strategy for mitigating ferroptosis-induced skin damage, warranting further investigation in advanced skin models and clinical applications.