Effects of extracellular calcium on viability and

Authors: Shaowen Cheng Wei Wang Zhongqin Lin Ping Zhou Xiaolei Zhang Wei Zhang Qingyu Chen Dongquan Kou Xiaozhou Ying Yue Shen Xiaojie Cheng Ziming Yu Lei Peng Chuanzhu Lu

Publish Date: 2013/06/08

Volume: 26, Issue: 3, Pages: 114-120

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Abstract

Bone marrow stromal cells BMSCs have been extensively used for tissue engineering However the effect of Ca2+ on the viability and osteogenic differentiation of BMSCs has yet to be evaluated To determine the dosedependent effect of Ca2+ on viability and osteogenesis of BMSCs in vitro BMSCs were cultured in calciumfree DMEM medium supplemented with various concentrations of Ca2+ 0 1 2 3 4 and 5 mM from calcium citrate Cell viability was analyzed by MTT assay and osteogenic differentiation was evaluated by alkaline phosphatase ALP assay Von Kossa staining and realtime PCR Ca2+ stimulated BMSCs viability in a dosedependent manner At slightly higher concentrations 4 and 5 mM in the culture Ca2+ significantly inhibited the activity of ALP on days 7 and 14 P  001 or P  005 significantly suppressed collagen synthesis P  001 or P  005 and significantly elevated calcium deposition P  001 and mRNA levels of osteocalcin P  001 or P  005 and osteopontin P  001 or P  005 Therefore elevated concentrations of extracellular calcium may promote cell viability and latestage osteogenic differentiation but may suppress earlystage osteogenic differentiation in BMSCsThe authors thank all the staff in the Laboratory of Orthopaedic Research Institute and Scientific Research Center of Second Affiliated Hospital of Wenzhou Medical College This work was supported by grants from the National Natural Science Foundation of China 30800220/C100201 Each author certifies that his or her institution approved the animal protocol for this investigation and that all investigations were conducted in conformity with ethical principles of research

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