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.

Apoptotic effects of cigarette smoke extracts on mouse embryonic stem cells via oxidative stress

“…It has also been reported that oxidative stress may result in changes in the expression of cell cycle‐related proteins, including cyclin D1, p21, and p27 . Another study also suggested that the decrease in proliferation depends on the production of ROS . Based on the results of this previous study and our study, we suggest that the inhibition of cell proliferation by CSE observed in the present study might have been because of oxidative stress.…”

Clinical, radiological, and gene expression analyses in smokers and non‐smokers, Part 2: RCT on the late healing phase of osseointegration

“…Whereas reduced bone mineral density has been reported in smokers, a careful search of the literature reveals that actual knowledge of the bone ultrastructure in smokers is missing. Another interpretation, supported by in vitro studies, is that smoking affects bone in a graded manner, necessitating both distinct anamnestic details and the implementation of an objective marker of smoking, such as the concentration of nicotine. Furthermore, as suggested by the present study, the maxilla and mandible respond differently to the constituents of tobacco smoke.…”

Effects of nicotine in the presence and absence of vitamin E on morphology, viability and osteogenic gene expression in MG-63 osteoblast-like cells

“…Evidence shows that nicotine down-regulates the expression of osteogenic genes [14]. The three main genes involved in bone metabolism include alkaline phosphatase (ALP), osteocalcin (OC) and bone sialoprotein (BSP) [3,15]. Alkaline phosphatase plays an important role in mineralization of collagenous bone matrix and is related to the activity of osteoblasts [16].…”

“…Alkaline phosphatase plays an important role in mineralization of collagenous bone matrix and is related to the activity of osteoblasts [16]. Osteocalcin [3] and BSP [17] are the main non-collagenous proteins in bone, which are involved in regulation of the mineralization process. Expression of OC occurs at a late stage of osteogenic differentiation and is a characteristic marker of mature cells of osteoblastic lineage [18].…”

“…The major effects of smoking on periodontal tissues include bone resorption, development of periodontal pockets and eventual tooth loss [2]. Nicotine as the main toxic component of cigarette smoke increases oxidative stress due to the formation of free radicals [3]. Animal studies have indicated that oxidative stress due to the effects of nicotine affects bone metabolism and delays bone healing [4][5][6][7].…”