Category Archives: Nicotine

Nicotine Not Responsible for Negative Effects of Smoking on Bone Density in Rats


The effect of long-term nicotine exposure on bone mineral density and oxidative stress in female Swiss Albino rats.

To evaluate the effect of long-term low or high-dose nicotine exposure on bone mass via measuring bone mineral density (BMD) and oxidant-antioxidant status markers.
Thirty-five female Swiss Albino rats weighing 70 ± 10 g were divided as the control group (n = 12), low-dose nicotine group (n = 12) and high-dose nicotine group (n = 11). While the control group was given only normal drinking water, the low-dose nicotine group had 0.4 mg/kg per day and the high-dose nicotine group, 6.0 mg/kg per day of nicotine added to their water for the period of 1 year. BMD was determined with X-ray absorptiometry of lumbar vertebra, corpus femoris, proximal and distal femur. To evaluate oxidant-antioxidant status malondialdehyde (MDA) levels, superoxide dismutase (SOD) and catalase (CAT) activities were determined.
When comparing the nicotine groups and controls, neither BMD nor oxidant-antioxidant status markers showed any statistically significant difference. In comparison to the controls, 12 months of high-dose oral nicotine exposure did not have a significant effect on BMD and low-dose nicotine exposure led to a statistically insignificant increase in BMD.
Contrary to common belief, the results of this study show that nicotine is not responsible for the decrease in BMD leading to osteoporosis frequently seen in smokers. However, there is a need to explore the other harmful materials in tobacco which may be responsible for the alterations seen in BMD of smokers.

Turan V, Mizrak S, Yurekli B, Yilmaz C…
Arch. Gynecol. Obstet. Feb 2013
PMID: 22955292

Moderate Alcohol is Good; Caffeine with Low Calcium is Bad


To drink or not to drink: how are alcohol, caffeine and past smoking related to bone mineral density in elderly women?

To determine relationship between alcohol, caffeine, past smoking and bone mineral density of different skeletal sites in elderly women, accounting for other biological and life-style variables.
A cross-sectional study in 136 Caucasian women, mean +/- SD age 68.6 +/- 7.1 years, all healthy and free of medications affecting bones, including estrogen. Bone mineral density (BMD) of multiple skeletal regions and body composition were measured by dual X-ray absorptiometry. Serum vitamin D (25-OHD) and parathyroid hormone (PTH) were analyzed and used as confounders. Calcium (Ca) intake was assessed by food frequency questionnaire. Alcohol and caffeine consumption was assessed by questionnaires determining frequency, amount and source of each. There were no current smokers, but the history of smoking was recorded, including number of years and packages smoked/day. Past physical activity was assessed by Allied Dunbar National Fitness Survey and used as confounder. Statistical significance was considered at p <or= 0.05.
In the correlational analysis, alcohol was positively associated with spine BMD (r = 0.197, p = 0.02), 25-OHD and negatively with PTH. Smoking was negatively related to Ca intake, 25(OH)D and number of reproductive years. In subgroup (stratified by Ca intake) and multiple regression analyses, alcohol (average approximately 0.5-1 drinks/day or approximately 8 g alcohol/day) was favorably associated with BMD of spine and total body. Caffeine (average approximately 2.5 6-fl oz cups/day or 200-300 mg caffeine/day) had negative association with most of the skeletal sites, which was attenuated with higher Ca intake (>or=median, 750 mg/day). The past smokers who smoked on average 24 years of approximately 1 pack cigarettes/day had lower BMD in total body, spine and femur than never-smokers when evaluated in subgroup analyses, and the association was attenuated in participants with >or=median Ca intake. There was no significant association between past smoking and BMD of any skeletal site in multiple regression analyses.
The results support the notion that consumption of small/moderate amount of alcohol is positively, while caffeine and past smoking are negatively associated with most of the skeletal sites, which might be attenuated with Ca intake above 750 mg/day.

Ilich JZ, Brownbill RA, Tamborini L, Crncevic-Orlic Z
J Am Coll Nutr Dec 2002
PMID: 12480799

It is interesting how many things are bad when calcium is low. There is some evidence that high protein, caffeine, and sodium are all bad for bones only when calcium is low. Otherwise, they all may be moderately good for bones when calcium is high.


Vitamin E with Tocotrienols is an Anabolic Bone Agent in Nicotine Treated Rats


Effects of palm vitamin e on bone-formation-related gene expression in nicotine-treated rats.

The study determines the effects of palm vitamin E on the gene expression of bone-formation-related genes in nicotine-treated rats. Male rats were divided into three groups: normal saline olive oil (NSO), nicotine olive oil (NO), and nicotine palm vitamin E (NE). The treatment was carried out in 2 phases. During the first 2 months, the NSO group received normal saline while the NO and NE groups received nicotine 7 mg/kg, 6 days a week, intraperitoneally. The following 2 months, normal saline and nicotine administration was stopped and was replaced with oral supplementation of olive oil for the NSO and NO groups and oral supplementation of palm vitamin E (60 mg/kg) for the NE group. Both femurs were harvested to determine the gene expression of bone morphogenetic protein-2 (BMP-2), Osterix (OSX), and Runt-related transcription factor 2 (RUNX2). Nicotine significantly downregulated the gene expression. This effect was reversed by palm vitamin E treatment. In conclusion, palm vitamin E may play a role in osteoblast differentiation and can be considered as an anabolic agent to treat nicotine-induced osteoporosis.

Abukhadir SS, Mohamed N, Makpol S, Muhammad N
Evid Based Complement Alternat Med 2012
PMID: 23049610 | Free Full Text

They used “Palm Vitamin E” in this study. Palm typically contains a high amount of Tocotrienols.

Vitamin E is an important fat-soluble vitamin with antioxidant properties. Of the two types of vitamin E, tocopherol is found in vegetable oils such as soy oil whereas tocotrienol is abundant in palm oil [7]. Previous studies have confirmed the beneficial effects of palm-oil-derived cotrcotrienol in several experimental osteoporosis; ovariectomized rats [8], steroid-induced rats [9], ferric-nitrilotriacetate-induced rats [10], and nicotine-induced rats [11, 12]. Furthermore, recent study has shown that supplementation of palm vitamin E, especially gamma isomer, can improve bone structural and biomechanical properties of normal male rats. Therefore, palm vitamin E has the potential to be used as an anabolic agent [13].


Tocotrienols Stimulate Bone Formation in Rats


Beneficial effects of tocotrienol and tocopherol on bone histomorphometric parameters in sprague-dawley male rats after nicotine cessation.

This study was conducted to determine the effectiveness of three forms of vitamin E supplements following nicotine treatment on bone histomorphometric parameters in an adult male rat model. Rats were divided into seven groups: baseline (B, killed without treatment), control (C, normal saline for 4 months), nicotine (N, nicotine for 2 months), nicotine cessation (NC), tocotrienol-enhanced fraction (TEF), gamma-tocotrienol (GTT), and alpha-tocopherol (ATF). Treatments for the NC, TEF, GTT, and ATF groups were performed in two phases. For the first 2 months they were given nicotine (7 mg/kg), and for the following 2 months nicotine administration was stopped and treatments with respective vitamin E preparations (60 mg/kg) were commenced except for the NC group, which was allowed to recover without treatment. Rats in the N and NC groups had lower trabecular bone volume, mineral appositional rate (MAR), and bone formation rate (BFR/BS) and higher single labeled surface and osteoclast surface compared to the C group. Vitamin E treatment reversed these nicotine effects. Both the TEF and GTT groups, but not the ATF group, had a significantly higher trabecular thickness but lower eroded surface (ES/BS) than the C group. The tocotrienol-treated groups had lower ES/BS than the ATF group. The GTT group showed a significantly higher MAR and BFR/BS than the TEF and ATF groups. In conclusion, nicotine induced significant bone loss, while vitamin E supplements not only reversed the effects but also stimulated bone formation significantly above baseline values. Tocotrienol was shown to be slightly superior compared to tocopherol. Thus, vitamin E, especially GTT, may have therapeutic potential to repair bone damage caused by chronic smoking.

Hermizi H, Faizah O, Ima-Nirwana S, Ahmad Nazrun S…
Calcif. Tissue Int. Jan 2009
PMID: 19020790

Vitamin E Reverses Nicotine-Induced Bone Resorption in Rats


Vitamin E reversed nicotine-induced toxic effects on bone biochemical markers in male rats.

Vitamin E is beneficial in restoring bone histomorphometric parameters in nicotine-treated rats. This study determined the effectiveness of 3 forms of vitamin E in restoring bone metabolism in nicotine-treated rats.
Thirty-five male Sprague-Dawley rats were divided into 5 groups: (1) control (C), (2) nicotine cessation (NC), (3) α-tocopherol (ATF), (4) tocotrienol-enhanced fraction (TEF) and (5) γ-tocotrienol (GTT). Treatment was carried out for 4 months. The control group was administered normal saline and olive oil throughout the treatment period while treatment for groups 2-5 was performed in 2 phases. In the first phase, the groups received nicotine 7 mg/kg intraperitoneally for 2 months. The following 2 months, group 2 received normal saline and olive oil while groups 3-5 received ATF, TEF or GTT, 60 mg/kg orally. Pre-treatment and post-treatment serum was collected for bone biochemical marker measurement using the ELISA method.
Nicotine increased serum bone-resorbing cytokines (interleukin-1 and interleukin-6) and the bone resorption marker pyridinoline (PYD) while reducing the bone formation marker osteocalcin after 2 months of nicotine treatment. The parameters failed to improve after nicotine was stopped for 2 months. Supplementation with the 3 forms of vitamin E improved the parameters, i.e. reduced the cytokines and pyridinoline as well as increased the osteocalcin. In addition, the TEF and GTT groups had a higher level of osteocalcin than the control group.
Nicotine impaired bone metabolism and cessation of nicotine treatment did not reverse the effects. Vitamin E, especially the tocotrienols, restored bone metabolism that was impaired due to nicotine.

Norazlina M, Hermizi H, Faizah O, Nazrun AS…
Arch Med Sci Aug 2010
PMID: 22371792 | Free Full Text

Coffee Delays Bone Repair in Rats


Effects of cigarette smoke inhalation and coffee consumption on bone formation and osseous integration of hydroxyapatite implant.

The present study aims to assess the effects of cigarette smoke inhalation and/or coffee consumption on bone formation and osseous integration of a dense hydroxyapatite (DHA) implant in rats. For this study, 20 male rats were divided into four groups (n = 5): CT (control) group, CE (coffee) group, CI (cigarette) group and CC (coffee + cigarette) group. During 16 weeks, animals in the CI group were exposed to cigarette smoke inhalation equivalent to 6 cigarettes per day; specimens in the CE group drank coffee as liquid diet; and rats in the CC group were submitted to both substances. In the 6th week a 5 mm slit in the parietal bone and a 4 mm slit in the tibia were performed on the left side: the former was left open while the latter received a DHA implant. As soon as surgeries were finished, the animals returned to their original protocols and after 10 weeks of exposure they were euthanised (ethically sacrificed) and the mentioned bones collected for histological processing. Data showed that exposure to cigarette smoke inhalation and coffee consumption did not interfere in weight gain and that solid and liquid diet consumption was satisfactory. Rats in the CC group showed a decrease in bone neoformation around the tibial DHA implant (31.8 ± 2.8) as well as in bone formation in the parietal slit (28.6 ± 2.2). On their own, cigarette smoke inhalation or coffee consumption also led to diminished bone neoformation around the implant and delayed the bone repair process in relation to the CT group. However, reduction in the bone repair process was accentuated with exposure to both cigarette smoke inhalation and coffee consumption in this study.

Andrade AR, Sant’Ana DC, Mendes JA, Moreira M…
Braz J Biol Feb 2013
PMID: 23644799 | Free Full Text

Quercetin Protects Human Osteoblasts Cells Exposed to Cigarette Smoke


Quercetin protects primary human osteoblasts exposed to cigarette smoke through activation of the antioxidative enzymes HO-1 and SOD-1.

Smokers frequently suffer from impaired fracture healing often due to poor bone quality and stability. Cigarette smoking harms bone cells and their homeostasis by increased formation of reactive oxygen species (ROS). The aim of this study was to investigate whether Quercetin, a naturally occurring antioxidant, can protect osteoblasts from the toxic effects of smoking. Human osteoblasts exposed to cigarette smoke medium (CSM) rapidly produced ROS and their viability decreased concentration- and time-dependently. Co-, pre- and postincubation with Quercetin dose-dependently improved their viability. Quercetin increased the expression of the anti-oxidative enzymes heme-oxygenase- (HO-) 1 and superoxide-dismutase- (SOD-) 1. Inhibiting HO-1 activity abolished the protective effect of Quercetin. Our results demonstrate that CSM damages human osteoblasts by accumulation of ROS. Quercetin can diminish this damage by scavenging the radicals and by upregulating the expression of HO-1 and SOD-1. Thus, a dietary supplementation with Quercetin could improve bone matter, stability and even fracture healing in smokers.

Braun KF, Ehnert S, Freude T, Egaña JT…
ScientificWorldJournal 2011
PMID: 22203790 | Free Full Text

Horny Goat Weed Stimulates Bone, Inhibits Turnover and Resorption in Smoking Rats


Effect of epimedium pubescen flavonoid on bone mineral status and bone turnover in male rats chronically exposed to cigarette smoke.

Epimedii herba is one of the most frequently used herbs in formulas that are prescribed for the treatment of osteoporosis in China and its main constituent is Epimedium pubescen flavonoid (EPF). However, it is unclear whether EPF during chronic exposure to cigarette smoke may have a protective influence on the skeleton. The present study investigated the effect of EPF on bone mineral status and bone turnover in a rat model of human relatively high exposure to cigarette smoke.
Fifty male Wistar rats were randomized into five groups: controls, passive smoking groups and passive smoking rats administered EPF at three dosage levels (75, 150 or 300 mg/kg/day) in drinking water for 4 months. A rat model of passive smoking was prepared by breeding male rats in a cigarette-smoking box. Bone mineral content (BMC), bone mineral density (BMD), bone turnover markers, bone histomorphometric parameters and biomechanical properties were examined.
Smoke exposure decreased BMC and BMD, increased bone turnover (inhibited bone formation and stimulated its resorption), affected bone histomorphometry (increased trabecular separation and osteoclast surface per bone surface; decreased trabecular bone volume, trabecular thickness, trabecular number, cortical thickness, bone formation rate and osteoblast surface per bone surface), and reduced mechanical properties. EPF supplementation during cigarette smoke exposure prevented smoke-induced changes in bone mineral status and bone turnover.
The results suggest that EPF can prevent the adverse effects of smoke exposure on bone by stimulating bone formation and inhibiting bone turnover and bone resorption.

Gao SG, Cheng L, Li KH, Liu WH…
BMC Musculoskelet Disord 2012
PMID: 22713117 | Free Full Text