Necrotic and apoptotic cells serve as nuclei for calcification on osteoblastic differentiation of human mesenchymal stem cells in vitro
Necrotic and apoptotic cells serve as nuclei for calcification on osteoblastic differentiation of human mesenchymal stem cells in vitro
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DOI：
10.1002/cbf.2974
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发表时间：
2014-01-01
影响因子：
3.6
通讯作者：
Sasaki, Junzo
中科院分区：
文献类型：
--
作者：
Fujita, Hirofumi;Yamamoto, Masanao;Sasaki, Junzo
细胞死亡与病理性钙化关系密切，如动脉粥样硬化中的血管钙化。然而，细胞死亡在成骨细胞系钙化中的作用尚未详细阐明。本研究探讨在体外成骨培养条件下，细胞死亡是否参与了人骨髓间充质干细胞向成骨细胞分化的钙化过程。人骨髓间充质干细胞在成骨培养过程中出现细胞凋亡和坏死，细胞死亡先于钙化。在这种培养条件下，细胞内活性氧的产生、染色质凝聚和碎裂以及caspase-3的激活增加。泛型半胱氨酸氨基转移酶抑制剂(Z-VAD-FMK)和抗氧化剂(TIron和N-乙酰半胱氨酸)可抑制成骨细胞诱导的细胞死亡和钙化。此外，添加坏死性死亡细胞或其膜部分可显著促进钙化。成骨培养的自发性死亡细胞和外源性增加的坏死细胞周围有钙沉积。在成骨细胞培养中，光动力处理诱导局部细胞死亡导致共同定位的钙化。这些结果表明，在hMSC的成骨培养过程中，出现了坏死和凋亡的细胞死亡，表明在体外hMSC的成骨分化过程中，成骨细胞系的坏死和凋亡细胞都是钙化的细胞核。版权所有(C)2013 John Wiley&Sons，Ltd.
A close relationship between cell death and pathological calcification has recently been reported, such as vascular calcification in atherosclerosis. However, the roles of cell death in calcification by osteoblast lineage have not been elucidated in detail. In this study, we investigated whether cell death is involved in the calcification on osteoblastic differentiation of human bone marrow mesenchymal stem cells (hMSC) under osteogenic culture in vitro. Apoptosis and necrosis occurred in an osteogenic culture of hMSC, and cell death preceded calcification. The generation of intracellular reactive oxygen species, chromatin condensation and fragmentation, and caspase-3 activation increased in this culture. A pan-caspase inhibitor (Z-VAD-FMK) and anti-oxidants (Tiron and n-acetylcysteine) inhibited osteogenic culture-induced cell death and calcification. Furthermore, calcification was significantly promoted by the addition of necrotic dead cells or its membrane fraction. Spontaneously dead cells by osteogenic culture and exogenously added necrotic cells were surrounded by calcium deposits. Induction of localized cell death by photodynamic treatment in the osteogenic culture resulted in co-localized calcification. These findings show that necrotic and apoptotic cell deaths were induced in an osteogenic culture of hMSC and indicated that both necrotic and apoptotic cells of osteoblast lineage served as nuclei for calcification on osteoblastic differentiation of hMSC in vitro. Copyright (c) 2013 John Wiley & Sons, Ltd.