title: Dose-dependent effects of nicotine on proliferation and differentiation of human bone marrow stromal cells and the antagonistic action of vitamin C.
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.
authors: Department of Orthopaedic Surgery, The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang Province 325000, People's Republic of China.
authors: Yue
authors: Y
authors: Shen
authors: Hai-xiao
authors: HX
authors: Liu
authors: Xiao-zhou
authors: XZ
authors: Ying
authors: Shi-zhou
authors: SZ
authors: Yang
authors: Peng-fei
authors: PF
authors: Nie
authors: Shao-wen
authors: SW
authors: Cheng
authors: Wei
authors: W
authors: Wang
authors: Xiao-jie
authors: XJ
authors: Cheng
authors: Lei
authors: L
authors: Peng
authors: Hua-zi
authors: HZ
authors: Xu
mesh_terms: D000328:Adult; D000368:Aged; D000975:Antioxidants / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D055396:Bone Morphogenetic Protein 2 / Q000096:biosynthesis; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004336:Drug Antagonism; D005260:Female; D005731:Ganglionic Stimulants / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D008875:Middle Aged; D009538:Nicotine / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D010012:Osteogenesis / Q000187:drug effects*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D000975:Antioxidants; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D005731:Ganglionic Stimulants; D009538:Nicotine; D001205:Ascorbic Acid