Department of Basic and Oral Biology, Bone Research Lab, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
Unit of Basic Oral Investigation-UIBO, School of Dentistry, Universidad El Bosque, Bogotá, Colombia.
Mol Biol Rep. 2021 Jan;48(1):13-20. doi: 10.1007/s11033-020-06096-w. Epub 2021 Jan 17.
Natural substances with antioxidant effects may benefit prevention and treatment of people with or prone to bone diseases after menopause, such as osteoporosis. This study aimed to evaluate the in vitro effect of preadministration of yerba mate extract (YM) in the metabolism of MC3T3-E1 osteoblasts exposed to hydrogen peroxide (HO). The cells (MC3T3-E1) were cultured in 24-well plates with the concentration of 1 μg/mL yerba mate extract dissolved in culture medium throughout the culture period. Four hours before each experiment, 400 μmol/L HO was added per well to simulate oxidative stress. There were evaluated cell adhesion and proliferation, in situ detection of alkaline phosphatase (ALP), mineralized nodules, and immunolocalization of osteocalcin (OCN), bone sialoprotein (BSP) and alkaline phosphatase (ALP) proteins. The results showed that YM preadministration to HO exposition significatively increased cell adhesion after 3 days as well as proliferation and in situ ALP detection after 10 and 7 days respectively, when compared to HO group. Besides, staining of OCN and BSP proteins was less intense and scattered in poor spread cells with cytoskeletal changes in HO group when compared to control and YM HO group. ALP staining was restrained to intracellular regions and similar in all experimental groups. Our results suggest that preadministration of yerba mate extract may prevent deleterious effects in the morphology and functional activity of osteoblasts exposed to HO, which could enable the maintenance of extracellular matrix in the presence of oxidative stress.
具有抗氧化作用的天然物质可能有益于预防和治疗绝经后或易患骨病的人群，如骨质疏松症。本研究旨在评估在暴露于过氧化氢（HO）的 MC3T3-E1 成骨细胞代谢中预先给予马黛茶提取物（YM）的体外作用。细胞（MC3T3-E1）在 24 孔板中培养，整个培养期间培养基中含有 1μg/mL 的马黛茶提取物。在每次实验前 4 小时，每孔加入 400μmol/L 的 HO 以模拟氧化应激。评估细胞黏附与增殖、原位碱性磷酸酶（ALP）检测、矿化结节形成以及骨钙素（OCN）、骨涎蛋白（BSP）和碱性磷酸酶（ALP）蛋白的免疫定位。结果表明，与 HO 组相比，YM 预处理可显著增加 HO 暴露后第 3 天的细胞黏附，第 10 天和第 7 天的增殖和原位 ALP 检测。此外，与对照组和 YM+HO 组相比，HO 组中 OCN 和 BSP 蛋白的染色较弱且分散，细胞铺展不良且细胞骨架发生变化。ALP 染色局限于细胞内区域，在所有实验组中相似。我们的结果表明，预先给予马黛茶提取物可能会防止暴露于 HO 的成骨细胞形态和功能活性的有害影响，这可能使细胞外基质在氧化应激存在的情况下得以维持。
