Eggshell-derived Nanoparticles Accelerate Wound Healing

Abstract

The chronic wounds (a silent epidemic) and its adverse impact is a serious global health problem. Normal wound healing is a complex, and processes through a series of four interconnected, partially overlapping stages: hemostasis, inflammation, proliferation, and remodelling. Angiogenesis (new vasculature produced from pre-existing blood vessels), the proliferative phase, serves pivotal function in wound healing. In our earlier published literature, we reported the pro-angiogenic properties of eggshell-derived nanoparticles (ES-NP). Considering the central role of angiogenesis in regulating wound healing, the present study systematically demonstrates the therapeutic efficacy of ES-NP through several in vitro assays and in vivo experiments. Wound healing involves highly regulated process of cellular events, in which keratinocytes serve as key mediators by driving re-epithelialization and barrier restoration. Initially, several in vitro experiments, including MTT assay, thymidine incorporation assay, cell cycle nes& apoptosis assay, ROS generation, and immunocytochemical analysis are performed in human keratinocytes (HaCaT) to support wound healing properties of ES-NP. Later, we have validated wound healing activity of ES-NP in pre-clinical mouse (C57BL/6) model where treatment of ES-NP significantly accelerates wound closure compared to untreated control. The results are supported by histopathology (H & E staining), immunohistochemistry (Ki-67 and CD31/PECAM), and Masson’s trichome staining (collagen deposition) of mice skin tissue. Altogether our findings suggest that ES-NP could be a potential candidate for the treatment of wounds (acute and chronic) and other diseases, where angiogenesis plays an important role.