利用软骨细胞片和松质骨构建骨软骨样组织及评价。

Tissue Engineering Part A Pub Date : 2021-02-01 Epub Date: 2020-09-18 DOI:10.1089/ten.TEA.2020.0107
Sopita Wongin, Rapeepat Narkbunnam, Saranatra Waikakul, Pojchong Chotiyarnwong, Thanyawan Aresanasuwan, Sittiruk Roytrakul, Kwanchanok Viravaidya-Pasuwat
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引用次数: 9

摘要

人体软骨细胞片在靶区的操作经常导致撕裂,因为它们是薄而脆弱的。本研究以人松质骨为支撑材料,制备软骨细胞片状松质骨组织,并对其性能进行评价。利用细胞片技术,将人软骨细胞构建成三层软骨细胞片,显示出软骨形成的特性。将软骨细胞片移植到松质骨上后,培养7天后,软骨细胞片-松质骨组织的顶部区域呈现光滑的表面形貌,无细胞片漂浮。同时进行II型胶原蛋白(COL2A1)和纤维连接蛋白(FN1)的免疫荧光染色。使用散弹枪蛋白质组学分析,在软骨细胞片、松质骨和软骨细胞片-松质骨组织中观察到与细胞粘附、细胞外基质(ECM)组织、细胞-底物连接组装和整合素介导的细胞粘附相关的蛋白质。在软骨细胞薄片中发现了三个整合素成员,包括整合素β4 (ITGB4)、ITGB6和ITGB8。仅在软骨细胞片和软骨细胞片-松质骨组织中发现ITGB8。在48 h内,单个细胞的平均迁移速度较低,但不影响软骨细胞片的结构和成软骨特性。对迁移细胞中丝状肌动蛋白(F-actin)细胞骨架的染色也可以更好地了解细胞骨架通过ITGB8与粘附分子之间的动态通信,这可能在软骨细胞片的附着和软骨ECM的合成中起关键作用。因此,我们建议将松质骨作为支撑材料构建软骨细胞片松质骨组织,用于骨软骨病变的潜在治疗。我们提出了一种将人软骨细胞片置于松质骨上构建骨软骨样组织的方法。在COL2A1表达的松质骨上,固定的软骨细胞片和单个细胞迁移的低平均速度表明,松质骨是一种合适的支撑材料。此外,基于itgb8介导的丝状肌动蛋白重排介导的软骨细胞片在松质骨上的粘附的细胞机制为改善骨软骨样组织的构建和预测骨关节炎治疗中的修复机制提供了更好的理解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Construction and Evaluation of Osteochondral-Like Tissue Using Chondrocyte Sheet and Cancellous Bone.

The manipulation of human chondrocyte sheets in target areas frequently results in their tearing because they are thin and fragile. In this study, human cancellous bones were used as a supporting material to create chondrocyte sheet-cancellous bone tissues, and their properties were evaluated. Using cell sheet technology, human chondrocytes were constructed into triple-layered chondrocyte sheets that displayed chondrogenic properties. After transferring the chondrocyte sheets onto cancellous bones, the top area of the chondrocyte sheet-cancellous bone tissues exhibited a smooth surface topography without cell sheet floating within 7 days of culture. The immunofluorescence staining of collagen type II (COL2A1) and fibronectin (FN1) was also performed and examined. Using the shotgun proteomic analysis, the proteins associated with cell adhesion, extracellular matrix (ECM) organization, cell-substrate junction assembly, and cell adhesion mediated by integrin were observed in the chondrocyte sheets, cancellous bones, and chondrocyte sheet-cancellous bone tissues. Three integrin members, including integrin β4 (ITGB4), ITGB6, and ITGB8, were found in the chondrocyte sheets. Only ITGB8 was found in the chondrocyte sheets and chondrocyte sheet-cancellous bone tissues. During 48 h, the mean velocity of the individual cell migration was low, which did not affect the structure and chondrogenic properties of the chondrocyte sheets. Staining of the filamentous actin (F-actin) cytoskeleton in the migratory cells also provided a better understanding of the dynamic communication between the cell cytoskeleton and adhesion molecules through ITGB8, which may play a key role in the attachment of the chondrocyte sheets and the synthesis of the cartilage ECM. Therefore, we suggest that cancellous bone could be used as a supporting material to construct chondrocyte sheet-cancellous bone tissues for potential treatment of osteochondral lesions. Impact Statement We proposed a method to construct an osteochondral-like tissue by placing human chondrocyte sheets onto cancellous bone. The stationary chondrocyte sheets and the low mean velocity of the individual cell migration on the cancellous bone with the expression of COL2A1 indicated that the cancellous bone served as an appropriate supporting material. Moreover, the cellular mechanism for the adhesion of the chondrocyte sheets on the cancellous bone based on ITGB8-mediated adhesion through the rearrangement of filamentous actin provided a better understanding to improve the construction of osteochondral-like tissues, and to predict the repair mechanism in osteoarthritis therapy.

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Tissue Engineering Part A
Tissue Engineering Part A CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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