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Journal of Cellular Biochemistry

Dose-dependent effects of nicotine on proliferation and differentiation of human bone marrow stromal cells and the antagonistic action of vitamin C

Authors

  • Yue Shen,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    2. The First/Second College of Clinical Medicine Department, Wenzhou Medical College, University-Town in Wenzhou, Zhejiang Province 325035, People's Republic of China
    3. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang Province 310000, People's Republic of China
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  • Hai-xiao Liu,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    2. The First/Second College of Clinical Medicine Department, Wenzhou Medical College, University-Town in Wenzhou, Zhejiang Province 325035, People's Republic of China
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  • Xiao-zhou Ying,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    2. The First/Second College of Clinical Medicine Department, Wenzhou Medical College, University-Town in Wenzhou, Zhejiang Province 325035, People's Republic of China
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  • Shi-zhou Yang,

    1. The First/Second College of Clinical Medicine Department, Wenzhou Medical College, University-Town in Wenzhou, Zhejiang Province 325035, People's Republic of China
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  • Peng-fei Nie,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    2. The First/Second College of Clinical Medicine Department, Wenzhou Medical College, University-Town in Wenzhou, Zhejiang Province 325035, People's Republic of China
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  • Shao-wen Cheng,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
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  • Wei Wang,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
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  • Xiao-jie Cheng,

    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    2. The First/Second College of Clinical Medicine Department, Wenzhou Medical College, University-Town in Wenzhou, Zhejiang Province 325035, People's Republic of China
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  • Lei Peng,

    Corresponding author
    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    2. Department of Trauma Center, The Affiliated Hospital of Hainan Medical College, 31 Longhua Road, Haikou, Hainan Province 570206, People's Republic of China
    • Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China.
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  • Hua-zi Xu

    Corresponding author
    1. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China
    • Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China.
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Abstract

A range of biological and molecular effects caused by nicotine are considered to effect bone metabolism. Vitamin C functions as a biological antioxidant. This study was to evaluate the in vitro effects of nicotine on human bone marrow stromal cells and whether Vitamin C supplementation show the antagonism action to high concentration nicotine. We used CCK-8, alkaline phosphatase (ALP) activity assay, Von Kossa staining, real-time polymerase chain reaction and Western Blot to evaluate the proliferation and osteogenic differentiation. The results indicated that the proliferation of BMSCs increased at the concentration of 50, 100 ng/ml, got inhibited at 1,000 ng/ml. When Vitamin C was added, the OD for proliferation increased. For ALP staining, we found that BMSCs treated with 50 and 100 ng/ml nicotine showed a higher activity compared with the control, and decreased at the 1,000 ng/ml. Bone morphogenetic protein-2 (BMP-2) expression and the calcium depositions decreased at 100 and 1,000 ng/ml nicotine, while the addition of Vitamin C reversed the down regulation. By real-time PCR, we detected that the mRNA expression of collagen type I (COL-I) and ALP were also increased in 50 and 100 ng/ml nicotine groups (P < 0.05), while reduced at 1,000 ng/ml (P < 0.05). When it came to osteocalcin (OCN), the changes were similar. Taken all together, it is found that nicotine has a two-phase effect on human BMSCs, showing that low level of nicotine could promote the proliferation and osteogenic differentiation while the high level display the opposite effect. Vitamin C could antagonize the inhibitory effect of higher concentration of nicotine partly. J. Cell. Biochem. 114: 1720–1728, 2013. © 2013 Wiley Periodicals, Inc.

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