Decellularized Persian Walnut Leaf (Juglans regia) Scaffold as a Potential Wound Dressing graft: An Experimental Study

Mehdi Kian ^1,2 Seyedeh-Sara Hashemi ^1,3* Amin Derakhshanfar ^1* Gholam-Hossein Darya ^1,2* Zahra Shahhosein ^3* Mohammad-Jamal Saharkhiz ^4,5*

• ¹ Department of Comparative Biomedical Sciences, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Fars, Iran
• ² Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran
• ³ Burn and Wound Healing Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
• ⁴ Department of Horticultural Science, School of Agriculture, Shiraz University, Shiraz, Fars, Iran
• ⁵ Medicinal Plants Processing Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

Wound dressings often fall short of providing the multifunctional capabilities required for optimal wound healing, such as promoting cell migration, proliferation, and tissue regeneration.Decellularization of plant tissues has gained attention as a potential source of biomaterials for tissue engineering applications due to the possession of favorable characteristics, including preexisting vascular networks, interconnected porous structure, efficient water transport and retention, high surface area, and a diverse range of mechanical properties. This study investigates the feasibility of using decellularized walnut leaves (DWL) as a novel scaffold for wound dressing in a mice model of excisional wounds. The decellularization and bleaching processes were carried out using various chemical agents. DNA and protein quantification and hematoxylin and eosin staining revealed successful removal of cells in DWL. Scanning electron microscopy (SEM) indicated that the normal structure of walnut leaves was preserved after the chemical decellularization. Chemical characterization by furrier transforms infrared and Raman spectroscopy showed the remaining some bioactive molecules and components in the structure of DWL. Comparing tensile strength and surface roughness parameters, surface wettability, swelling, and porosity properties of native and DWL indicated no statistical differences between them. SEM analysis indicated that human mesenchymal stem cells excellently attach and proliferate on the DWL. Additionally, the biocompatibility and potential of DWL scaffolds to accelerate wound closure and enhance histopathological score, collagen deposition, and epithelial thickness were observed in a mice model of the excisional wound. In conclusion, DWL has a promising potential for applicating as a skin wound dressing.