This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: [10.1002/jcb.24512].
Dose-dependent effects of nicotine on proliferation and differentiation of human bone marrow stromal cells and the antagonistic action of vitamin C†
Copyright © 2013 Wiley Periodicals, Inc.
- Accepted manuscript online: 5 FEB 2013 08:17AM EST
- Manuscript Accepted: 24 JAN 2013
- Manuscript Revised: 25 NOV 2012
- Manuscript Received: 19 MAR 2012
- Project of Science and Technology Department of Zhejiang Province. Grant Number: 2010C34006/2010C34G2090013
- National Natural Science Foundation of China. Grant Number: 31060135/C100302
- Project of Health Department in Hainan Province. Grant Number: 2011-34
- Science-Technology Hall of Hainan province. Grant Number: GJXM201102
- Cited By
- Vitamin C;
- human BMSCs;
- Osteogenic Differentiation
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, 100ng/ml, got inhibited at 1000 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 1000 ng/ml. Bone morphogenetic protein-2 (BMP-2) expression and the calcium depositions decreased at 100 and 1000ng/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 1000 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. © 2013 Wiley Periodicals, Inc.