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Effects of extracellular calcium on viability and osteogenic differentiation of bone marrow stromal cells in vitro

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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 dose-dependent effect of Ca2+ on viability and osteogenesis of BMSCs in vitro, BMSCs were cultured in calcium-free 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 real-time PCR. Ca2+ stimulated BMSCs viability in a dose-dependent 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 < 0.01 or P < 0.05), significantly suppressed collagen synthesis (P < 0.01 or P < 0.05), and significantly elevated calcium deposition (P < 0.01) and mRNA levels of osteocalcin (P < 0.01 or P < 0.05) and osteopontin (P < 0.01 or P < 0.05). Therefore, elevated concentrations of extracellular calcium may promote cell viability and late-stage osteogenic differentiation, but may suppress early-stage osteogenic differentiation in BMSCs.

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Acknowledgments

The 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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Authors and Affiliations

  1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, 109 Xue Yuan Xi Road, Wenzhou, 325000, China

    Wei Wang, Xiaolei Zhang, Wei Zhang, Qingyu Chen, Dongquan Kou, Xiaozhou Ying, Yue Shen & Xiaojie Cheng

  2. Trauma Center of the Affiliated Hospital of Hainan Medical College, 31 Long Hua Road, Haikou, 571100, China

    Shaowen Cheng, Ziming Yu, Lei Peng & Chuanzhu Lu

  3. Department of Oncology, The First Affiliated Hospital of Zhejiang University, Hangzhou, China

    Ping Zhou

  4. Department of Orthopaedic Surgery, The Hospital of Integrated Traditional Chinese and Western Medicine of Wenzhou, Wenzhou, China

    Zhongqin Lin

Authors
  1. Shaowen Cheng
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  2. Wei Wang
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  3. Zhongqin Lin
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  4. Ping Zhou
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  5. Xiaolei Zhang
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  6. Wei Zhang
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  7. Qingyu Chen
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  8. Dongquan Kou
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  9. Xiaozhou Ying
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  10. Yue Shen
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  11. Xiaojie Cheng
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  12. Ziming Yu
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  13. Lei Peng
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  14. Chuanzhu Lu
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Corresponding authors

Correspondence to Lei Peng or Chuanzhu Lu.

Additional information

S. Cheng and W. Wang contributed equally to this work.

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Cheng, S., Wang, W., Lin, Z. et al. Effects of extracellular calcium on viability and osteogenic differentiation of bone marrow stromal cells in vitro. Human Cell 26, 114–120 (2013). https://doi.org/10.1007/s13577-012-0041-8

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  • DOI: https://doi.org/10.1007/s13577-012-0041-8

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