Formulation, Evaluation, Factorial Optimization, and In-Silico Study of Eplerenone Loaded Pectin Nanoparticles: A New Approach to the Proliferation of Human Skin Fibroblasts for Wound Healing

Objective

Repurposing Eplerenone (EPL) for wound healing through in silico molecular docking to explore its therapeutic potential beyond its traditional use (antihypertensive drug). In this study, for a first time we investigated the potential of EPL- loaded pectin nanoparticles to enhance human skin proliferation.

Methods

Ionotropic gelation method was used for preparing these EPL-loaded PN using calcium chloride as a cross-linker. Design® Expert software was used to utilize a full factorial 2³ design, where the selected factors were the pectin concentration (X₁), EPL amount (X₂), and pectin: calcium chloride concentration (X₃). The selected responses were the EPL entrapment efficiency percentage, particle size and zeta potential for determining the optimum formula for further studies such as in-vitro release studies, transmission electron microscopy (TEM), X-ray diffraction, cell culture study and in-vitro evaluation on human skin fibroblast proliferation cell.

Results

The selected system demonstrated high entrapment efficiency (80.56 ± 0.62%), a particle size of 509.1 ± 45.5 nm, and good zeta potential value (-21.73 ± 2.1 mV). A spherical morphology with no aggregation using TEM and amorphization of the crystalline drug in X-ray diffraction were obtained. The Sulfo-Rhodamine B assay was applied for its effect on cell viability on human skin cells, and the selected concentration was 50 μg/mL.

Conclusions

The wound healing assay on human skin fibroblasts using scratch wound technique showed the superiority of EPL on wound healing; suggesting its promising effect in enhancing skin healing and tissue regeneration. Moreover, pectin activity as a wound healing accelerator, promoting a synergistic effect.

Graphical Abstract