title: Necrotic and apoptotic cells serve as nuclei for calcification on osteoblastic differentiation of human mesenchymal stem cells in vitro.
abstract: 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.
authors: Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
authors: Hirofumi
authors: H
authors: Fujita
authors: Masanao
authors: M
authors: Yamamoto
authors: Tetsuya
authors: T
authors: Ogino
authors: Hirotsugu
authors: H
authors: Kobuchi
authors: Naoko
authors: N
authors: Ohmoto
authors: Eriko
authors: E
authors: Aoyama
authors: Takashi
authors: T
authors: Oka
authors: Tohru
authors: T
authors: Nakanishi
authors: Keiji
authors: K
authors: Inoue
authors: Junzo
authors: J
authors: Sasaki
mesh_terms: D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt / Q000494:pharmacology; D000111:Acetylcysteine / Q000494:pharmacology; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D002113:Calcification, Physiologic; D053148:Caspase 3 / Q000378:metabolism; D061945:Caspase Inhibitors / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D042002:Chromatin Assembly and Disassembly; D004789:Enzyme Activation; D006801:Humans; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D009336:Necrosis; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D000975:Antioxidants; D061945:Caspase Inhibitors; D017382:Reactive Oxygen Species; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt; D053148:Caspase 3; D000111:Acetylcysteine
keywords: apoptosis; calcification; mesenchymal stem cell; necrosis; osteoblast; reactive oxygen species