Ceccarelli Gabriele, Bloise Nora, Mantelli Melissa, Gastaldi Giulia, Fassina Lorenzo, De Angelis Maria Gabriella Cusella, Ferrari Davide, Imbriani Marcello, Visai Livia
Department of Public Health, Neuroscience, and Experimental & Forensic Medicine, University of Pavia , Pavia, Italy . ; Center for Tissue Engineering (C.I.T.), University of Pavia , Pavia, Italy .
Biores Open Access. 2013 Aug;2(4):283-94. doi: 10.1089/biores.2013.0016.
Human mesenchymal stem cells (MSCs) are a promising candidate cell type for regenerative medicine and tissue engineering applications. Exposure of MSCs to physical stimuli favors early and rapid activation of the tissue repair process. In this study we investigated the in vitro effects of pulsed electromagnetic field (PEMF) treatment on the proliferation and osteogenic differentiation of bone marrow MSCs (BM-MSCs) and adipose-tissue MSCs (ASCs), to assess if both types of MSCs could be indifferently used in combination with PEMF exposure for bone tissue healing. We compared the cell viability, cell matrix distribution, and calcified matrix production in unstimulated and PEMF-stimulated (magnetic field: 2 mT, amplitude: 5 mV) mesenchymal cell lineages. After PEMF exposure, in comparison with ASCs, BM-MSCs showed an increase in cell proliferation (p<0.05) and an enhanced deposition of extracellular matrix components such as decorin, fibronectin, osteocalcin, osteonectin, osteopontin, and type-I and -III collagens (p<0.05). Calcium deposition was 1.5-fold greater in BM-MSC-derived osteoblasts (p<0.05). The immunofluorescence related to the deposition of bone matrix proteins and calcium showed their colocalization to the cell-rich areas for both types of MSC-derived osteoblast. Alkaline phosphatase activity increased nearly 2-fold (p<0.001) and its protein content was 1.2-fold higher in osteoblasts derived from BM-MSCs. The quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis revealed up-regulated transcription specific for bone sialoprotein, osteopontin, osteonectin, and Runx2, but at a higher level for cells differentiated from BM-MSCs. All together these results suggest that PEMF promotion of bone extracellular matrix deposition is more efficient in osteoblasts differentiated from BM-MSCs.
人间充质干细胞（MSCs）是再生医学和组织工程应用中一种很有前景的候选细胞类型。将MSCs暴露于物理刺激有利于组织修复过程的早期快速激活。在本研究中，我们调查了脉冲电磁场（PEMF）处理对骨髓间充质干细胞（BM-MSCs）和脂肪组织间充质干细胞（ASCs）增殖和成骨分化的体外影响，以评估这两种类型的MSCs是否都能无差别地与PEMF暴露联合用于骨组织愈合。我们比较了未受刺激和PEMF刺激（磁场：2 mT，振幅：5 mV）的间充质细胞系中的细胞活力、细胞基质分布和钙化基质产生。PEMF暴露后，与ASCs相比，BM-MSCs的细胞增殖增加（p<0.05），细胞外基质成分如核心蛋白聚糖、纤连蛋白、骨钙素、骨连接蛋白、骨桥蛋白以及I型和III型胶原蛋白的沉积增强（p<0.05）。BM-MSC来源的成骨细胞中的钙沉积增加了1.5倍（p<0.05）。与骨基质蛋白和钙沉积相关的免疫荧光显示，这两种类型的MSC来源的成骨细胞中它们在富含细胞的区域共定位。碱性磷酸酶活性增加了近2倍（p<0.001），其蛋白含量在BM-MSCs来源的成骨细胞中高1.2倍。定量逆转录聚合酶链反应（qRT-PCR）分析显示，骨唾液蛋白、骨桥蛋白、骨连接蛋白和Runx2的转录特异性上调，但BM-MSCs分化的细胞上调水平更高。所有这些结果表明，PEMF促进骨细胞外基质沉积在BM-MSCs分化的成骨细胞中更有效。
