Background/Objectives: Hepcidin is a negative regulator of iron absorption that is released by hepatocytes. It is one of the main contributors to hypoferremia and anemia in inflammatory and oncological disorders that are mediated by the proinflammatory cytokine IL-6/STAT3 pathway. Ferulic acid (FA) is a phenolic compound with pleiotropic biological activities, including anti-inflammatory activity. However, its effect on hepcidin secretion is still unknown. Thus, this study aimed to explore the impact of FA on hepcidin levels and the underlying mechanism. Methods: HepG2 cells were treated with different log percentages of FA, and their viability was determined via the MTT assay. The relative expression of IL-6 and HAMP in treated and untreated cells was measured via qRT-PCR, and the protein levels of hepcidin, IL-6 and STAT3 were measured using ELISA. Results: The MTT test showed an inverse relationship between FA concentrations and HepG2 cell proliferation; FA’s IC₅₀ value was 0.07669%. The expression levels of IL-6 and HAMP were significantly increased in HepG2 cells following 24 h of culture with 4 μg/mL LPS. Meanwhile, the addition of FA significantly decreased the relative expression levels of these two genes and the secretion levels of IL-6, STAT3 and hepcidin compared to the cells treated with LPS alone. Conclusions: Overall, these findings show that FA modifies inflammatory pathways, affecting hepcidin levels via the IL-6/STAT3 pathway. Thus, this suggests FA as a potential therapeutic agent against the hypoferremia and anemia developed due to dysregulated hepcidin levels in diseases such as inflammatory and oncological disorders.

Hyperlipidemia is a major risk factor for cardiovascular diseases and is associated with complications such as atherosclerosis and tendon injury. Though atorvastatin reduces cholesterol, genetic variants (CYP2D6-4, SULT1A1, CYP2C192) affect its response. These genetic variations influence atorvastatin metabolism, thereby affecting its therapeutic effectiveness. To advance personalized therapeutic drug monitoring and improve lipid profile management, this study aims to develop a robust and LC-MS/MS method for quantifying atorvastatin levels in human plasma. Additionally, to investigate the influence of genetic polymorphisms - particularly CYP2D6-4-on plasma concentrations of atorvastatin in patients with hyperlipidemia. Ethical approval for the study was obtained from the appropriate institutional review boards, and written informed consent was obtained from all participants. Atorvastatin was measured using LC-MS/MS. PCR-based methods were used for genotyping. Statistical analyses were performed to evaluate relationships between plasma atorvastatin levels and genetic variants. The LC-MS/MS method demonstrated excellent linearity, accuracy, precision, and stability, for the quantification of atorvastatin in human plasma. Higher atorvastatin concentrations were tied to CYP2D6-4. Furthermore, the study validated the analytical method for consistent and reliable measurement of atorvastatin levels in clinical samples. This study successfully developed and validated a straightforward and reliable LC-MS/MS method for quantifying atorvastatin levels in human plasma. Significant CYP2D64 - atorvastatin links highlight the value of pharmacogenetic dosing. Integrating pharmacogenetics - especially in the Jordanian population - may enhance the safety, efficacy, and individualization of atorvastatin therapy.

Background: Diabetes-associated wound healing is frequently impaired due to persistent hyperglycemia, which causes vascular dysfunction and neuropathy, thereby delaying tissue repair. Objectives: In this study, hyaluronic acid and protamine nanoparticles (HA-PRO-NPs) were developed and optimized to encapsulate disulfiram (DSF) and coumarin (COM) for potential application in diabetic wound healing. Methods: The nanoparticles were prepared using a simple ionic interaction method and characterized for their particle size, zeta potential, morphology and encapsulation efficiency. Then, HA-PRO-NPs were incorporated into a hydrogel that contains carboxymethyl cellulose (CMC) and propylene glycol (PG). The hydrogel formulations were evaluated for swelling behavior, spreadability and DSF and COM release kinetics. Results: The prepared formulations were safe against human dermal fibroblasts (HDFs), with cell viability remaining above 70% over the concentration ranges tested. In the scratch assay, HA-PROT-DSF-COM-NPs improved HDF migration, achieving approximately 81% wound closure. An in vivo study was conducted using streptozotocin-induced diabetic mice. Topical application of Gel-HA-PROT-DSF-COM-NPs promoted wound closure. The study monitored wound contraction on Days 0, 3, 7, and 12. By Day 12, topical administration of the dual-cargo system (Gel-HA-PROT-DSF-COM NPs, n = 6) achieved 78.2 ± 4.1% wound closure. This was significantly higher than the single-drug formulation (Gel-HA-PROT-DSF-NPs: 59.5 ± 4.8%) and the untreated Control (Diabetic) group (55.4 ± 5.2%, p < 0.01), while the non-diabetic Control group (non diabetic) reached 97.6 ± 3.5% closure. Semi-quantitative histopathological evaluation showed that the Gel-HA-PROT-DSF-COM NPs achieved a healing score of 4/5, characterized by optimized epidermal regeneration with preserved adnexal structures and moderate dermal remodeling. Conclusions: In conclusion, Gel-HA-PROT-DSF-COM-NPs demonstrated promising potential as a topical formulation for improving wound healing under diabetic conditions.