Optimization of A Rapid Spinach Decellularization Technique using Tween-20 with Antibacterial and Wound Healing Properties

Abstract

Abstract Creating acellular constructs with vascularized structure from animal and plant tissue is one of the well-known strategies for scaffold assembly. One of these approaches is decellularization, and the used process for this method varies according to the nature of the tissue and its composition. The most common method is the chemical decellularization of plants. This method employs various high doses of detergents, primarily Triton X-100, sodium dodecyl sulfate (SDS), and sodium hypochlorite (SH). In this study, detergents, which are available in most of the laboratories were compared to develop new methods of spinach decellularization. Tween-20, SDS and SH in minimal concentrations were used to decellularize spinach leaves as an acellular plant matrix for tissue engineering. The samples divided in two different temperature conditions, and decellularization was proven by different analyses such as DNA and protein extraction and quantification and hematoxylin and eosin staining. The antibacterial activity was assessed to choose the most potential group. Two groups with the lowest DNA and protein amounts and high vasculature were chosen for the in vitro experiment. MTT and Scratch assay were used to evaluate the biocompatibility and ability of the bioconstructs to promote wound healing on fibroblasts. The results show that the Tween-20 decellularized samples at concentrations of 1% at 21°C and 37°C had the best morphology and clear vascularization with a high biocompatibility. Also, all of the scaffold samples are very good at killing S. aureus bacteria, which means they can help keep wounds from getting infected while they heal. This indicates they could be used as a scaffold for tissue engineering purposes.