Department of Bioengineering, Izmir Institute of Technology (IZTECH), 35430 Izmir, Turkey.
Department of Bioengineering, Izmir Institute of Technology (IZTECH), 35430 Izmir, Turkey.
Int J Biol Macromol. 2024 Jun;270(Pt 1):131856. doi: 10.1016/j.ijbiomac.2024.131856. Epub 2024 Apr 30.
Biomacromolecules derived from natural sources offer superior biocompatibility, biodegradability, and water-holding capacity, which make them promising scaffolds for tissue engineering. Psyllium seed has gained attention in biomedical applications recently due to its gel-forming ability, which is provided by its polysaccharide-rich content consisting mostly of arabinoxylan. This study focuses on the extraction and gelation of Psyllium seed hydrocolloid (PSH) in a single-step water-based protocol, and scaffold fabrication using freeze-drying method. After characterization of the scaffold, including morphological, mechanical, swelling, and protein adsorption analyses, 3D cell culture studies were done using NIH-3 T3 fibroblast cells on PSH scaffold, and cell viability was assessed using Live/Dead and Alamar Blue assays. Starting from day 1, high cell viability was obtained, and it reached 90 % at the end of 15-day culture period. Cellular morphology on PSH scaffold was monitored via SEM analysis; cellular aggregates then spheroid formation were observed throughout the study. Collagen Type-I and F-actin expressions were followed by immunostaining revealing a 9- and 10-fold increase during long-term culture. Overall, a single-step and non-toxic protocol was developed for extraction and gelation of PSH. Obtained results unveiled that PSH scaffold provided a favorable 3D microenvironment for cells, holding promise for further tissue engineering applications.
天然来源的生物大分子具有优越的生物相容性、可生物降解性和保水能力，因此它们是组织工程有前途的支架材料。由于其富含多糖的成分，主要由阿拉伯木聚糖组成，具有形成凝胶的能力，因此榆科植物种子在生物医学应用中引起了关注。本研究专注于在单步水基方案中提取和胶凝榆科植物种子水胶体（PSH），并使用冷冻干燥法制造支架。对支架进行了包括形态学、力学、溶胀和蛋白质吸附分析在内的特性表征后，使用 NIH-3T3 成纤维细胞在 PSH 支架上进行了 3D 细胞培养研究，并使用 Live/Dead 和 Alamar Blue 测定法评估了细胞活力。从第 1 天开始，获得了高细胞活力，在 15 天培养期结束时达到 90%。通过 SEM 分析监测 PSH 支架上的细胞形态；整个研究过程中观察到细胞聚集然后形成球体。通过免疫染色追踪胶原蛋白 I 型和 F-肌动蛋白的表达，显示在长期培养过程中分别增加了 9 倍和 10 倍。总体而言，开发了一种用于提取和胶凝 PSH 的单步无毒方案。获得的结果表明，PSH 支架为细胞提供了有利的 3D 微环境，有望进一步应用于组织工程。
