Accelerating wound healing rates with Cucurbita pepo leaf extract loaded electrospun poly(methyl methacrylate)/Halloysite/Chitosan/CaCO₃ composite nanofibers through in vitro and in vivo assessments

Abstract

Cucurbita pepo leaf extract (CPE) was incorporated into electrospun poly(methyl methacrylate)/ halloysite/chitosan/CaCO₃ (PMMA/Hal/CS/CaCO₃) composite nanofibers to develop a novel biomaterial for accelerating wound healing rates. The different nanofibrous scaffolds were successfully fabricated and characterized through scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). SEM analysis revealed uniform, smooth nanofibers, while FTIR and XRD confirmed the integration of CPE into nanofiber matrix, indicating an amorphous structure and effective dispersion of the extract. In vitro agar well-diffusion and antibiofilm assays revealed that the optimized formulation exhibited potent antimicrobial activity against wound-associated pathogens. The nanofibers composite based on PMMA provides structural stability, while Hal enhances mechanical properties and cell adhesion. Additionally, CS contributes to biodegradability and moisture retention, both of which are essential for effective wound healing. Furthermore, the inclusion of CPE offers additional antimicrobial benefits, further promoting the wound healing process. In vivo studies confirmed the wound-healing potential of the optimal loaded scaffold, with a 99.9% healing rate achieved by day 14. In vivo studies further demonstrated that CPE-loaded nanofibers significantly enhanced wound closure compared to untreated controls, likely due to the synergistic effects between CPE and PMMA/Hal/CS/CaCO₃ matrix. Histological analysis revealed increased macrophage infiltration, neovascularization, and fibroblast proliferation by day 7. Overall, these findings underscore the potential of incorporating natural bioactive compounds into electrospun nanofibers to develop biocompatible, eco-friendly antimicrobial materials for advanced wound-healing applications.