软骨素诱导脂肪干细胞外泌体增强脂肪干细胞的软骨分化能力。
Kartogenin Induced Adipose-Derived Stem Cell Exosomes Enhance the Chondrogenic Differentiation Ability of Adipose-Derived Stem Cells.
机构信息
Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China.
Department of Orthopedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
出版信息
Dis Markers. 2022 Aug 29;2022:6943630. doi: 10.1155/2022/6943630. eCollection 2022.
OBJECTIVE
The objective of this study is to explore the effect of kartogenin (KGN-)-pretreated adipose-derived stem cell-derived exosomes (ADSC-EXOs) on the chondrogenic differentiation ability of ADSCs.
METHODS
Adipose-derived stem cells (ADSCs) were treated with different doses of KGN, and exosomes (EXOs) were extracted. EXOs were then identified using an electron microscope (EM), nanoparticle tracking analyzer, nanoparticle tracking analysis software, and exosomal protein markers. EXOs were labeled with the fluorescent dye PKH67 and their uptake by cells was evaluated. A cell counting kit-8 (CCK-8) assay, flow cytometry, clonogenic assay, and a cell scratch assay were used to detect the abilities of proliferation, apoptosis, clone formation, and migration of ADSCs, respectively. Subsequently, Alcian blue staining and toluidine blue staining were used to detect the chondrogenic differentiation ability of ADSCs in each group. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot (WB) techniques were used to detect the expression of chondrogenic differentiation-related genes.
RESULTS
In this study, ADSCs and KGN-induced ADSC-EXOs were successfully extracted and isolated. EXOs and ADSCs coculturing results showed that KGN-induced ADSC-EXOs can significantly promote proliferation, clone formation, migration, and chondrogenic differentiation of ADSCs and inhibit apoptosis. In addition, KGN-induced ADSC-EXOs can increase the expression of chondrogenic-related genes in ADSCs (Aggrecan, Collagen III, Collagen II, and SOX9), and can significantly decrease the expression of chondrolysis-related genes (MMP-3, ADAMTS4, and ADAMTS5).
CONCLUSION
KGN-induced ADSC-EXOs can enhance the chondrogenic differentiation ability of ADSCs by promoting cell proliferation and migration while inhibiting cell apoptosis. KGN treatment can also increase the expression of chondrogenic differentiation-related genes and decrease the expression of chondrolysis-related genes. These results provide a new approach to cartilage repair and regeneration.
目的
本研究旨在探讨软骨素衍生基因（KGN）预处理脂肪来源干细胞衍生的外泌体（ADSC-EXOs）对 ADSC 软骨分化能力的影响。
方法
用不同剂量的 KGN 处理脂肪来源干细胞（ADSCs），提取外泌体（EXOs）。用电镜（EM）、纳米颗粒跟踪分析仪、纳米颗粒跟踪分析软件和外泌体蛋白标志物鉴定 EXOs。用荧光染料 PKH67 标记 EXOs，评估细胞摄取情况。用细胞计数试剂盒-8（CCK-8）检测细胞增殖能力，流式细胞术、克隆形成实验和细胞划痕实验分别检测细胞增殖、凋亡、克隆形成和迁移能力。随后，用茜素红染色和甲苯胺蓝染色检测各组 ADSC 的软骨分化能力。采用定量实时聚合酶链反应（qRT-PCR）和 Western blot（WB）技术检测软骨分化相关基因的表达。
结果
本研究成功提取并分离出 ADSC 和 KGN 诱导的 ADSC-EXOs。EXOs 和 ADSCs 共培养结果表明，KGN 诱导的 ADSC-EXOs 可显著促进 ADSC 的增殖、克隆形成、迁移和软骨分化，抑制凋亡。此外，KGN 诱导的 ADSC-EXOs 可增加 ADSC 中软骨相关基因（聚集蛋白聚糖、III 型胶原、II 型胶原和 SOX9）的表达，同时显著降低软骨降解相关基因（MMP-3、ADAMTS4 和 ADAMTS5）的表达。
结论
KGN 诱导的 ADSC-EXOs 通过促进细胞增殖和迁移，同时抑制细胞凋亡，增强 ADSC 的软骨分化能力。KGN 处理还可增加软骨分化相关基因的表达，降低软骨降解相关基因的表达。这些结果为软骨修复和再生提供了一种新方法。
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