Department of Orthodontics, School of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China.
Yunnan Key Laboratory of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China.
Biomed Res Int. 2021 Nov 27;2021:9962861. doi: 10.1155/2021/9962861. eCollection 2021.
Combining the advantages of static magnetic fields (SMF) and coculture systems, we investigated the effect of moderate-intensity SMF on the chondrogenesis and proliferation of mandibular bone marrow mesenchymal stem cells (MBMSCs) in the MBMSC/mandibular condylar chondrocyte (MCC) coculture system. The main aim of the present study was to provide an experimental basis for obtaining better cartilage tissue engineering seed cells for the effective repair of condylar cartilage defects in clinical practice.
MBMSCs and MCCs were isolated from SD (Sprague Dawley) rats. Flow cytometry, three-lineage differentiation, colony-forming assays, immunocytochemistry, and toluidine blue staining were used for the identification of MBMSCs and MCCs. MBMSCs and MCCs were seeded into the lower and upper Transwell chambers, respectively, at a ratio of 1 : 2, and exposed to a 280 mT SMF. MBMSCs were harvested after 3, 7, or 14 days for analysis. CCK-8 was used to detect cell proliferation, Alcian blue staining was utilized to evaluate glycosaminoglycan (GAG), and western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) detected protein and gene expression levels of SOX9, Col2A1 (Collagen Type II Alpha 1), and Aggrecan (ACAN).
The proliferation of MBMSCs was significantly enhanced in the experimental group with MBMSCs cocultured with MCCs under SMF stimulation relative to controls ( < 0.05). GAG content was increased, and SOX9, Col2A1, and ACAN were also increased at the mRNA and protein levels ( < 0.05).
Moderate-intensity SMF improved the chondrogenesis and proliferation of MBMSCs in the coculture system, and it might be a promising approach to repair condylar cartilage defects in the clinical setting.
结合静磁场（SMF）和共培养系统的优点，我们研究了中强度 SMF 对下颌骨髓间充质干细胞（MBMSCs）在 MBMSC/下颌髁突软骨细胞（MCC）共培养系统中的软骨生成和增殖的影响。本研究的主要目的是为获得更好的软骨组织工程种子细胞，以有效修复临床实践中的髁突软骨缺损提供实验依据。
从 SD（Sprague Dawley）大鼠中分离 MBMSCs 和 MCC。采用流式细胞术、三系分化、集落形成试验、免疫细胞化学和甲苯胺蓝染色鉴定 MBMSCs 和 MCC。将 MBMSCs 和 MCC 以 1：2 的比例分别接种到下室和上室的 Transwell 小室中，并暴露于 280 mT 的 SMF 中。在第 3、7 或 14 天收获 MBMSCs 进行分析。CCK-8 用于检测细胞增殖，阿尔辛蓝染色用于评估糖胺聚糖（GAG），Western blot 和实时定量聚合酶链反应（RT-qPCR）检测 SOX9、Col2A1（胶原 II 型 α1）和 Aggrecan（ACAN）的蛋白和基因表达水平。
与对照组相比，在 SMF 刺激下与 MCC 共培养的 MBMSCs 实验组的增殖明显增强（<0.05）。GAG 含量增加，SOX9、Col2A1 和 ACAN 的 mRNA 和蛋白水平也增加（<0.05）。
中强度 SMF 提高了共培养系统中 MBMSCs 的软骨生成和增殖能力，可能是修复临床髁突软骨缺损的一种有前途的方法。
