Chronic ethanol intake inhibits in vitro osteogenesis induced by osteoblasts differentiated from stem cells

• Published in: Journal of applied toxicology Vol. 28; no. 2; pp. 205 - 211
• Main Authors: Rosa, Maria L., Beloti, Marcio M., Prando, Natalia, Queiroz, Regina H. C., Oliveira, Paulo T. de, Rosa, Adalberto L.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-03-2008; Wiley
• Subjects: Alcoholism - complications; Alcoholism - metabolism; Alcoholism - pathology; Alcoholism - physiopathology; Alcoholism and acute alcohol poisoning; Alkaline Phosphatase - metabolism; Animals; Biological and medical sciences; bone; Bone Marrow Cells - drug effects; Bone Marrow Cells - metabolism; Bone Marrow Cells - pathology; cell culture; Cell Differentiation - drug effects; Cell Proliferation - drug effects; Cell Shape - drug effects; Cell Survival - drug effects; Cells, Cultured; ethanol; Ethanol - toxicity; Male; Medical sciences; osteoblast; Osteoblasts - drug effects; Osteoblasts - metabolism; Osteoblasts - pathology; Osteogenesis - drug effects; Osteoporosis - etiology; Osteoporosis - pathology; Osteoporosis - physiopathology; Protein Biosynthesis - drug effects; Rats; Rats, Wistar; stem cell; Stem Cells - drug effects; Stem Cells - metabolism; Stem Cells - pathology; Time Factors; Toxicology; Cell culture; Ethanol; Nutrition; Stem cell; In vitro; Feeding; Osteoarticular system; Alcoholic beverage; Chronic; Diet; Osteoblast; Inhibitor; Bone; Osteogenesis
• ISSN: 0260-437X; 1099-1263
• DOI: 10.1002/jat.1271

The study investigated whether chronic ethanol (ETH) intake and subsequent ETH exposure of cell cultures affects osteoblast differentiation by evaluating key parameters of in vitro osteogenesis. Rats were treated with 5–20% (0.85–3.43 mm) ETH, increasing by 5% per week for a period of 4 weeks (habituation), after which the 20% level was maintained for 15 days (chronic intake). Bone‐marrow stem cells from control (CONT) or ETH‐treated rats were cultured in osteogenic medium which was either supplemented (ETH) or not supplemented (CONT) with 1.3 mm ethanol. Thus, four groups relating to rat treatment/culture supplementation were evaluated: (1) CONT/CONT, (2) ETH/CONT, (3) CONT/ETH and (4) ETH/ETH. Cell morphology, proliferation and viability, total protein content, alkaline phosphatase (ALP) activity and bone‐like nodule formation were evaluated. Chronic ethanol intake significantly reduced both food and liquid consumption and body weight gain. No difference was seen in cell morphology among treatments. Cell number was affected at 7 and 10 days as follows: CONT/CONT = CONT/ETH < ETH/CONT = ETH/ETH. Doubling time between 3 and 10 days was greater in groups of CONT animals: ETH/ETH = ETH/CONT < CONT/ETH = CONT/CONT. Cell viability and ALP activity were not affected by either animal treatment or culture exposure to ethanol. At day 21, the total protein content was affected as follows: ETH/ETH = CONT/ETH < ETH/CONT = CONT/CONT. Bone‐like nodule formation was affected as follows: ETH/ETH < CONT/ETH < ETH/CONT < CONT/CONT. These results show that chronic ethanol intake, followed by the exposure of osteoblasts to ethanol, inhibited the differentiation of osteoblasts, as indicated by an increased proliferation rate and reduced bone‐like nodule formation. Copyright © 2007 John Wiley & Sons, Ltd.