Della Morte Elena, Notarangelo Maria Pina, Niada Stefania, Giannasi Chiara, Fortuna Federica, Cadelano Francesca, Lambertini Elisabetta, Piva Roberta, Brini Anna Teresa, Penolazzi Letizia
Laboratory of Biotechnological Applications, IRCCS Istituto Ortopedico Galeazzi, 20157, Milan, Italy.
Department of Neuroscience and Rehabilitation, University of Ferrara, 44121, Ferrara, Italy.
Calcif Tissue Int. 2025 Jan 7;116(1):26. doi: 10.1007/s00223-024-01335-9.
This study describes the potential of the conditioned medium (CM) from adipose-derived mesenchymal stromal cells (ASCs) to affect the response of bone cells and support bone remodeling. This was in particular assessed by an in vitro model represented by a 3D human osteoblast-osteoclast co-culture. It has been reported that the effects of ASCs are predominantly attributable to the paracrine effects of their secreted factors, that are present as soluble factors or loaded into extracellular vesicles. They may affect various biological processes, including bone turnover. Our interest was to provide further evidence to support ASC-CM as a promising cell-free therapeutic agent for the treatment of bone loss. ASC-CM was characterized using nanoparticle tracking analysis (NTA), cytofluorimetry, and proteomic analysis. Human osteoblasts (hOBs) from vertebral lamina were cultured with monocytes, as osteoclasts (hOCs) precursors, in a Rotary cell culture system for 14 days. Histochemical analysis was performed to evaluate the effect of ASC-CM on bone-specific markers such as tartrate-resistant acid phosphatase (TRAP), osteopontin (OPN), RUNX2, Collagen 1 (COL1), and mineral matrix. ASC-CM characterization confirmed the content of CD63/CD81/CD9 positive extracellular vesicles. Proteomic dataset considering bone-remodeling-related keywords identified 16 processes significantly enriched. The exposure of hOBs/hOCs aggregates to ASC-CM induced increase of OPN, COL I, and RUNX2, and significantly induced mineral matrix deposition, while significantly reducing TRAP expression. These data demonstrated that CM from ASCs contains a complex of secreted factors able to control either bone resorption or bone formation and requires further investigations to deeply analyze their potential therapeutic effects.
本研究描述了脂肪来源间充质基质细胞（ASC）的条件培养基（CM）影响骨细胞反应和支持骨重塑的潜力。这尤其通过以三维人成骨细胞-破骨细胞共培养为代表的体外模型进行评估。据报道，ASC的作用主要归因于其分泌因子的旁分泌作用，这些因子以可溶性因子形式存在或装载到细胞外囊泡中。它们可能影响包括骨转换在内的各种生物学过程。我们的兴趣在于提供进一步证据，以支持ASC-CM作为治疗骨质流失的一种有前景的无细胞治疗剂。使用纳米颗粒跟踪分析（NTA）、细胞荧光分析和蛋白质组学分析对ASC-CM进行了表征。将来自椎板的人成骨细胞（hOB）与作为破骨细胞（hOC）前体的单核细胞在旋转细胞培养系统中培养14天。进行组织化学分析以评估ASC-CM对骨特异性标志物如抗酒石酸酸性磷酸酶（TRAP）、骨桥蛋白（OPN）、RUNX2、胶原蛋白1（COL1）和矿物质基质的影响。ASC-CM表征证实了CD63/CD81/CD9阳性细胞外囊泡的含量。考虑到与骨重塑相关关键词的蛋白质组数据集确定了16个显著富集的过程。hOB/hOC聚集体暴露于ASC-CM会诱导OPN、COL I和RUNX2增加，并显著诱导矿物质基质沉积，同时显著降低TRAP表达。这些数据表明，ASC的CM含有能够控制骨吸收或骨形成的分泌因子复合物，需要进一步研究以深入分析其潜在的治疗效果。
