Ahn Sang-Wook, Kim Eun-Jung, Kim Mi Kyoung, Shin Sang-Hun, Kwon Jin-Ju
Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University, Dental and Life Science Institute, Yangsan, Republic of Korea.
Department of Dental Anesthesia and Pain Medicine, School of Dentistry, Pusan National University, Dental Research Institute, Yangsan, Republic of Korea.
J Dent Sci. 2025 Jan;20(1):212-219. doi: 10.1016/j.jds.2024.08.005. Epub 2024 Aug 19.
BACKGROUND/PURPOSE: Membrane-free stem cell components (MFSCCs) have been developed by removing cell membranes with antigens to overcome the limitations associated with cell-based therapies and isolate effective peptides. MFSCCs have been reported to have effects on oral infection sites. Chronic inflammatory diseases cause excessive bone resorption. This study investigated the effects of MFSCCs on osteoclast differentiation in the context of the high prevalence of inflammatory bone resorption.
Bone marrow macrophages (BMMs) were treated with macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand. Osteoclast differentiation was assessed based on the MFSCC concentrations. Tartrate-resistant acid phosphatase (TRAP)-stained mature osteoclasts and multinucleated cells derived from BMMs were analyzed using light microscopy. The messenger RNA (mRNA) expression levels of genes related to osteoclast differentiation were measured using real-time polymerase chain reaction (RT-PCR). The relative expression levels of the key transcription factors c-fos and nuclear factor of activated T cells (NFATc1) were determined using quantitative RT-PCR and western blotting.
After treatment with MFSCCs, the cell viability was similar, depending on the level of BMMs. As the MFSCC concentration increased, the number of TRAP-positive cells decreased. The mRNA and protein expression of cathepsin K, TRAP, dendritic cell-specific transmembrane protein, c-fos, and NFATc1 decreased as the MFSCC concentration increased.
Our findings demonstrate that MFSCCs suppress osteoclast differentiation by downregulating transcription factors, particularly, c-fos and NFATc1. Therefore, MFSCCs may serve as a conservative treatment option for chronic inflammatory bone resorption diseases of the oral cavity by suppressing excessive bone resorption.
背景/目的：通过去除带有抗原的细胞膜开发出无膜干细胞成分（MFSCCs），以克服基于细胞的疗法的局限性并分离有效的肽。据报道，MFSCCs对口腔感染部位有作用。慢性炎症性疾病会导致过度的骨吸收。本研究在炎症性骨吸收高发的背景下，研究了MFSCCs对破骨细胞分化的影响。
用巨噬细胞集落刺激因子和核因子κB受体激活剂配体处理骨髓巨噬细胞（BMMs）。根据MFSCC浓度评估破骨细胞分化。使用光学显微镜分析抗酒石酸酸性磷酸酶（TRAP）染色的成熟破骨细胞和源自BMMs的多核细胞。使用实时聚合酶链反应（RT-PCR）测量与破骨细胞分化相关基因的信使核糖核酸（mRNA）表达水平。使用定量RT-PCR和蛋白质印迹法测定关键转录因子c-fos和活化T细胞核因子（NFATc1）的相对表达水平。
用MFSCCs处理后，细胞活力相似，这取决于BMMs的水平。随着MFSCC浓度的增加，TRAP阳性细胞数量减少。随着MFSCC浓度的增加，组织蛋白酶K、TRAP、树突状细胞特异性跨膜蛋白、c-fos和NFATc1的mRNA和蛋白质表达降低。
我们的研究结果表明，MFSCCs通过下调转录因子，特别是c-fos和NFATc1来抑制破骨细胞分化。因此，MFSCCs可能通过抑制过度的骨吸收，作为口腔慢性炎症性骨吸收疾病的一种保守治疗选择。
