Liu Zhan, Zhou Haoyu, Wu Qingwei, Luo Tianhao, Tu Hanlin, Sa Guoliang, Yang Xuewen
State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, PR China.
Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Wuhan University, Wuhan, PR China.
Heliyon. 2024 Aug 13;10(17):e35972. doi: 10.1016/j.heliyon.2024.e35972. eCollection 2024 Sep 15.
An organoid culture system better recapitulates the cellular structure, function, and interaction between cells and the extracellular matrix (ECM) than a two-dimensional (2D) culture system. We here constructed a condylar cartilage organoid to explore the regulatory role of primary cilia. Similar to the natural condylar cartilage, the condylar cartilage organoid exhibited abundant ECM and comprised superficial, proliferative, and hypertrophic zones. Primary cilia in the condylar cartilage organoid were shorter on average than those in the 2D culture chondrocytes, but their average length was equivalent to those in the natural condylar cartilage. Notably, primary cilia in each zone of the condylar cartilage organoid had an average length similar to that of primary cilia in the natural condylar cartilage. According to transcriptomic and biochemical analyses, the expression of cilia-related genes and cilia-related Hedgehog (HH) signaling differed between the condylar cartilage organoid and 2D culture systems. knockdown promoted the protein levels of COL-Ⅹ, TRPV4, and HH signaling molecules in the condylar cartilage organoid, but decreased them in the 2D culture system. Notably, the protein levels of COL-Ⅹ, TRPV4, and HH signaling molecules increased in the superficial zone of the si condylar cartilage organoid compared with the condylar cartilage organoid. However, the protein levels of aforementioned molecules were not significantly different in proliferative and hypertrophic zones. Collectively, we successfully constructed the condylar cartilage organoid with a better tissue structure and abundant ECM. Moreover, the condylar cartilage organoid is more suitable for exploring primary cilia functions.
与二维（2D）培养系统相比，类器官培养系统能更好地重现细胞结构、功能以及细胞与细胞外基质（ECM）之间的相互作用。我们在此构建了髁突软骨类器官，以探索初级纤毛的调节作用。与天然髁突软骨相似，髁突软骨类器官表现出丰富的细胞外基质，并由表层、增殖层和肥大层组成。髁突软骨类器官中的初级纤毛平均长度比二维培养软骨细胞中的短，但其平均长度与天然髁突软骨中的相当。值得注意的是，髁突软骨类器官各层中的初级纤毛平均长度与天然髁突软骨中的相似。根据转录组学和生化分析，髁突软骨类器官和二维培养系统中纤毛相关基因的表达以及纤毛相关的刺猬（HH）信号传导存在差异。敲低髁突软骨类器官中促进了COL-Ⅹ、TRPV4和HH信号分子的蛋白水平，但在二维培养系统中降低了它们的水平。值得注意的是，与髁突软骨类器官相比，si髁突软骨类器官表层中COL-Ⅹ、TRPV4和HH信号分子的蛋白水平有所增加。然而，上述分子在增殖层和肥大层中的蛋白水平没有显著差异。总体而言，我们成功构建了具有更好组织结构和丰富细胞外基质的髁突软骨类器官。此外，髁突软骨类器官更适合探索初级纤毛的功能。
