Department of Biotechnology, Institute of Natural and Applied Sciences, Çukurova University, Balcalı, Adana, Türkiye.
Department of Translational Medicine, Faculty of Medicine, Cukurova University, Balcali, Adana, Türkiye.
J Biomed Mater Res B Appl Biomater. 2024 Oct;112(10):e35489. doi: 10.1002/jbm.b.35489.
Creating acellular vascularized constructs from animal and plant tissue is one of the well-known strategies for scaffold assembly. Decellularization takes an important position among these strategies. The most common method is chemical decellularization. This approach employs high concentrations of detergents, primarily Triton X-100, sodium dodecyl sulfate (SDS), and sodium hypochlorite (SH). In this work, novel techniques for decellularizing spinach were developed using detergents frequently utilized in laboratories. Spinach leaves were decellularized using Tween-20, SDS, and SH at low concentrations to generate an acellular plant matrix for tissue engineering. We measured the quantities of DNA and protein, as well as the decellularization using hematoxylin and eosin (H&E) staining. The biocompatibility and capacity of the biostructures to stimulate fibroblast wound healing were assessed using MTT and the Scratch assay. The antibacterial activity of the scaffolds was also tested against a gram-positive bacterium, Staphilococcus aureus, which is a common pathogen associated with wound healing. The best shape, evident vascularization, and good biocompatibility were seen in the Tween-20 decellularized samples at 1% concentration at 21°C and 37°C through the enhancement of cell proliferation and wound healing. In terms of antibacterial activity, all scaffold samples had a significant effect on Staphilococcus aureus, where the number of bacterial colonies in all six scaffold groups became zero after 4 h of treatment. The scaffolds also showed a 100% kill rate on Staphilococcus aureus, which could avoid wound infection during the repair process, and that can be suggested as a scaffold for tissue engineering applications and an important constituent for pharmacological activities.
从动物和植物组织中创建无细胞血管化构建体是支架组装的知名策略之一。脱细胞化在这些策略中占有重要地位。最常见的方法是化学脱细胞。这种方法采用高浓度的去污剂，主要是 Triton X-100、十二烷基硫酸钠（SDS）和次氯酸钠（SH）。在这项工作中，使用实验室中常用的去污剂开发了菠菜脱细胞的新技术。菠菜叶在低浓度的 Tween-20、SDS 和 SH 下脱细胞，以生成用于组织工程的无细胞植物基质。我们测量了 DNA 和蛋白质的数量，以及使用苏木精和曙红（H＆E）染色进行的脱细胞化。使用 MTT 和划痕测定法评估生物结构的生物相容性和刺激成纤维细胞伤口愈合的能力。还针对革兰氏阳性菌金黄色葡萄球菌测试了支架的抗菌活性，金黄色葡萄球菌是与伤口愈合相关的常见病原体。在 21°C 和 37°C 下，浓度为 1%的 Tween-20 脱细胞样品的最佳形状、明显的血管化和良好的生物相容性通过增强细胞增殖和伤口愈合得到了证明。在抗菌活性方面，所有支架样本对金黄色葡萄球菌都有显著影响，所有六个支架组中的细菌菌落数量在治疗 4 小时后均变为零。支架对金黄色葡萄球菌的杀灭率也达到 100%，可以避免修复过程中的伤口感染，这可以作为组织工程应用的支架和药理学活性的重要组成部分。
