Shi Linyan, Ura Kazuhiro, Takagi Yasuaki
Graduate School of Fisheries Sciences, Hokkaido University, 3-1-1 Minato-Cho, Hakodate, Hokkaido, 041-8611, Japan.
Faculty of Fisheries Sciences, Hokkaido University, 3-1-1 Minato-cho, Hakodate, Hokkaido, 041-8611, Japan.
Osteoarthr Cartil Open. 2024 Feb 27;6(2):100450. doi: 10.1016/j.ocarto.2024.100450. eCollection 2024 Jun.
Although type II collagen could have marked potential for developing cartilage tissue engineering (CTE) scaffolds, its erratic supply and viscous nature have limited these studies, and there are no studies on the use of marine-derived type II collagen fibrils for CTE scaffold materials. In this study, we aimed to generate a fibril-based, thin-layered scaffold from marine-derived type II collagen and investigate its chondrogenic potential.
Time-lapse observations revealed the cell adhesion process. The Cell Counting Kit-8 (CCK-8) assay, light microscopy, and scanning electron microscopy were performed to detect proliferation and filopodium morphology. Alcian blue staining was used to show the deposition of extracellular secretions, and qRT-PCR was performed to reveal the expression levels of chondrogenesis-related genes.
The cell adhesion speed was similar in both fibril-coated and control molecule-coated groups, but the cellular morphology, proliferation, and chondrogenesis activity differed. On fibrils, more elongated finer filopodia showed inter-cell communications, whereas the slower proliferation suggested an altered cell cycle. Extracellular secretions occurred before day 14 and continued until day 28 on fibrils, and on fibrils, the expression of the chondrogenesis-related genes (), (), (), and ( = 0.0049) was significantly upregulated on day 21.
Marine-derived type II collagen was, for the first time, fabricated into a fibril state. It showed rapid cellular affinity and induced chondrogenesis with extracellular secretions. We presented a new model for studying chondrogenesis and a potential alternative material for cell-laden CTE research.
尽管II型胶原蛋白在开发软骨组织工程（CTE）支架方面具有显著潜力，但其供应不稳定和粘性性质限制了这些研究，并且尚无关于将海洋来源的II型胶原纤维用于CTE支架材料的研究。在本研究中，我们旨在从海洋来源的II型胶原蛋白制备基于纤维的薄层支架，并研究其软骨生成潜力。
延时观察揭示了细胞粘附过程。进行细胞计数试剂盒-8（CCK-8）测定、光学显微镜和扫描电子显微镜以检测增殖和丝状伪足形态。阿尔辛蓝染色用于显示细胞外分泌物的沉积，qRT-PCR用于揭示软骨生成相关基因的表达水平。
纤维包被组和对照分子包被组的细胞粘附速度相似，但细胞形态、增殖和软骨生成活性不同。在纤维上，更多细长的细丝状伪足显示细胞间通讯，而较慢的增殖表明细胞周期改变。在纤维上，细胞外分泌物在第14天之前出现并持续到第28天，并且在纤维上，软骨生成相关基因（）、（）、（）和（ = 0.0049）的表达在第21天显著上调。
首次将海洋来源的II型胶原蛋白制成纤维状态。它显示出快速的细胞亲和力并通过细胞外分泌物诱导软骨生成。我们提出了一种研究软骨生成的新模型和一种用于载细胞CTE研究的潜在替代材料。
