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

Bo Han; William Fang; Zhi Yang; Yuntao Wang; Shuqing Zhao; Ba Xuan Hoang; C. Thomas Vangsness

doi:10.3390/biomedicines11123145

Biomedicines (Nov 2023)

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

Bo Han,
William Fang,
Zhi Yang,
Yuntao Wang,
Shuqing Zhao,
Ba Xuan Hoang,
C. Thomas Vangsness

Affiliations

Bo Han: Department of Surgery and Biomedical Engineering, Keck School of Medicine, University of Southern California, 1333 San Pablo St., BMT 302A, Los Angeles, CA 90089, USA
William Fang: Department of Orthopaedic Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089, USA
Zhi Yang: Department of Surgery and Biomedical Engineering, Keck School of Medicine, University of Southern California, 1333 San Pablo St., BMT 302A, Los Angeles, CA 90089, USA
Yuntao Wang: Department of Surgery and Biomedical Engineering, Keck School of Medicine, University of Southern California, 1333 San Pablo St., BMT 302A, Los Angeles, CA 90089, USA
Shuqing Zhao: Department of Surgery and Biomedical Engineering, Keck School of Medicine, University of Southern California, 1333 San Pablo St., BMT 302A, Los Angeles, CA 90089, USA
Ba Xuan Hoang: Department of Surgery and Biomedical Engineering, Keck School of Medicine, University of Southern California, 1333 San Pablo St., BMT 302A, Los Angeles, CA 90089, USA
C. Thomas Vangsness: Department of Orthopaedic Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089, USA

DOI: https://doi.org/10.3390/biomedicines11123145
Journal volume & issue: Vol. 11, no. 12
p. 3145

Abstract

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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.

Published in Biomedicines

ISSN: 2227-9059 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Biology (General)
Website: http://www.mdpi.com/journal/biomedicines

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