Enhancement of Chondrogenic Markers by Exosomes Derived from Cultured Human Synovial Fluid-Derived Cells: A Comparative Analysis of 2D and 3D Conditions

被引:5
作者
Han, Bo [1 ]
Fang, William [2 ]
Yang, Zhi [1 ]
Wang, Yuntao [1 ]
Zhao, Shuqing [1 ]
Hoang, Ba Xuan [1 ]
Vangsness, C. Thomas [2 ]
机构
[1] Univ Southern Calif, Keck Sch Med, Dept Surg & Biomed Engn, 1333 San Pablo St,BMT 302A, Los Angeles, CA 90089 USA
[2] Univ Southern Calif, Keck Sch Med, Dept Orthopaed Surg, Los Angeles, CA 90089 USA
关键词
exosome; chondrogenesis; synovial fluid-derived cells; 3D encapsulation; cartilage repair; MESENCHYMAL STEM-CELLS; CARTILAGE TISSUE; ARTICULAR CHONDROCYTES; BONE-MARROW; OSTEOARTHRITIS; REGENERATION; SCAFFOLDS;
D O I
10.3390/biomedicines11123145
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Objective: The goal of this pilot study was to investigate the effects of exosomes derived from synovial fluid-derived cells (SFDCs) cultured under normoxic conditions in a two-dimensional (2D) monolayer or encapsulated within a three-dimensional (3D) matrix for chondrogenic differentiation in vitro and cartilage defect repair in vivo. Design: Synovial fluid samples were obtained from three patients, and SFDCs were isolated and expanded either in a 2D monolayer culture or seeded within a transglutaminase cross-linked gelatin (Col-Tgel) to create a 3D gel culture. Exosomes derived from each environment were isolated and characterized. Then, their effects on cartilage-cell proliferation and chondrogenic differentiation were assessed using an in vitro organoid model, and their potential for enhancing cartilage repair was evaluated using a rat cartilage defect model. Results: SFDCs obtained from different donors reached a state of senescence after four passages in 2D culture. However, transferring these cells to a 3D culture environment mitigated the senescence and improved cell viability. The 3D-cultured exosomes exhibited enhanced potency in promoting chondrogenic differentiation, as evidenced by the increased expression of chondrogenic genes and greater deposition of cartilage-specific extracellular matrix. Furthermore, the 3D-cultured exosomes demonstrated superior effectiveness in enhancing cartilage repair and exhibited better healing properties compared to exosomes derived from a 2D culture. Conclusions: The optimized 3D culture provided a more favorable environment for the proliferation of human synovial cells and the secretion of exosomes compared to the 2D culture. The 3D-cultured exosomes exhibited greater potential for promoting chondrogenic gene expression in vitro and demonstrated improved healing properties in repairing cartilage defects compared to exosomes derived from the 2D culture.
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页数:15
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