Formulation and In Vitro Characterisation of Withaferin A-Loaded Liposomal Gels for the Topical Management of Chronic Inflammatory Skin Conditions

Chronic inflammatory skin conditions such as psoriasis, eczema, and acne are driven by sustained inflammation, oxidative stress, and impaired tissue repair. Current treatments often lead to adverse effects with prolonged use highlighting the need for safer, targeted alternatives. Withaferin-A, a bioactive compound derived from <i>Withania somnifera</i>, has demonstrated potent anti-inflammatory and antioxidant properties in various disease models. This study explored the potential of Withaferin-A liposome-loaded gels for topical delivery, testing their efficacy in an inflamed skin model. Withaferin-A liposomes were prepared using the ethanol injection method and incorporated into hydroxypropyl methylcellulose (HPMC) gels. <i>In vitro</i> release studies using a permeable insert system were used to compare release profiles of Withaferin-A from liposomal gels. Cytotoxicity was assessed via XTT assay on human umbilical vein endothelial cells (HUVEC) and human dermal fibroblasts (HDFa). Inflammation was induced with tumour necrosis factor-alpha (TNF-α), and anti-inflammatory effects measured using enzyme-linked immunosorbent assays for interleukin-6 (IL-6) and matrix metalloproteinase-9 (MMP9). Reactive oxygen species (ROS) levels were quantified using the DCFDA assay. Cytotoxicity studies using the XTT assay on HUVEC and HDFa cells revealed good biocompatibility at lower Withaferin-A concentrations (0-1 µM), though reduced viability was observed at 5 µM. <i>In vitro</i> release studies revealed sustained release of Withaferin-A from liposomal gels, with significantly slower release over 6 h compared to solution at 99.53% ± 3.47% and 48.87% ± 4.51% respectively. Anti-inflammatory effects were evaluated following TNF-α-induced inflammation, with Withaferin-A loaded gels significantly reducing IL-6 secretion in a dose-dependent manner in both HUVECs (38.90 ± 5.34 to 19.15 ± 3.56 pg/mL) and HDFa cells (40.05 ± 2.23 to 10.42 ± 2.02 pg/mL). Withaferin-A treatment also reduced ROS levels and lowered MMP-9 secretion in HDFa cells from 408.80 ± 13.05 pg/mL to 195.00 ± 7.55 pg/mL, indicating potential for tissue remodelling. In summary, WA-loaded liposomal gels demonstrated effective anti-inflammatory activity and sustained drug release while maintaining biocompatibility at therapeutic concentrations. These findings support their potential as a novel strategy for managing inflammatory skin diseases by improving drug bioavailability and promoting tissue repair.