Zumbo Bianca, Guagnini Benedetta, Medagli Barbara, Porrelli Davide, Turco Gianluca
Department of Medicine, Surgery and Health Sciences, University of Trieste, Piazza dell'Ospitale 1, 34129 Trieste, Italy.
Department of Life Sciences, University of Trieste, Via Alexander Fleming 31/B, 34127 Trieste, Italy.
J Funct Biomater. 2024 Aug 10;15(8):222. doi: 10.3390/jfb15080222.
Bone defects are a global health concern; bone tissue engineering (BTE) is the most promising alternative to reduce patient morbidity and overcome the inherent drawbacks of autograft and allograft bone. Three-dimensional scaffolds are pivotal in this field due to their potential to provide structural support and mimic the natural bone microenvironment. Following an already published protocol, a 3D porous structure consisting of alginate and hydroxyapatite was prepared after a gelation step and a freezing-drying step. Despite the frequent use of alginate in tissue regeneration, the biological inertness of this polysaccharide hampers proper cell colonization and proliferation. Therefore, the purpose of this work was to enhance the biological properties by promoting the interaction and adhesion between cells and biomaterial with the use of Fibronectin. This extracellular matrix protein was physically adsorbed on the scaffold, and its presence was evaluated with environmental scanning electron microscopy (eSEM) and the Micro-Bicinchoninic Acid (μBCA) protein assay. The MG-63 cell line was used for both static and dynamic (i.e., in bioreactor) 3D cell culturing on the scaffolds. The use of the bioreactor allowed for a better exchange of nutrients and oxygen and a better removal of cell catabolites from the inner portion of the construct, mimicking the physiological environment. The functionalized scaffolds showed an improvement in cell proliferation and colonization compared to non-functionalized ones; the effect of the addition of Fibronectin was more evident in the dynamic culturing conditions, where the cells clearly adhered on the surface of functionalized scaffolds.
骨缺损是一个全球性的健康问题；骨组织工程（BTE）是最有前景的替代方法，可降低患者发病率并克服自体骨移植和异体骨移植的固有缺点。三维支架在该领域至关重要，因为它们有潜力提供结构支撑并模拟天然骨微环境。按照已发表的方案，在凝胶化步骤和冷冻干燥步骤之后，制备了一种由藻酸盐和羟基磷灰石组成的三维多孔结构。尽管藻酸盐在组织再生中经常使用，但这种多糖的生物惰性阻碍了细胞的正常定植和增殖。因此，这项工作的目的是通过使用纤连蛋白促进细胞与生物材料之间的相互作用和粘附来增强生物学特性。这种细胞外基质蛋白被物理吸附在支架上，并通过环境扫描电子显微镜（eSEM）和微量双辛可宁酸（μBCA）蛋白测定法评估其存在情况。MG-63细胞系用于在支架上进行静态和动态（即在生物反应器中）三维细胞培养。使用生物反应器可以更好地交换营养物质和氧气，并更好地从构建体内部去除细胞分解代谢物，模拟生理环境。与未功能化的支架相比，功能化的支架在细胞增殖和定植方面有改善；在动态培养条件下，添加纤连蛋白的效果更明显，此时细胞明显粘附在功能化支架的表面。
