Annatto-derived tocotrienol stimulates osteogenic activity in preosteoblastic MC3T3-E1 cells: a temporal sequential study

• Published in: Drug design, development and therapy development and therapy, 2018-01, Vol.12, p.1715-1726
• Main Authors: Wan Hasan, Wan Nuraini, Abd Ghafar, Norzana, Chin, Kok-Yong, Ima-Nirwana, Soelaiman
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Alizarin; Alkaline phosphatase; Alkaline Phosphatase - genetics; Alkaline Phosphatase - metabolism; Animal models; Animals; Annatto; Biocompatibility; Bixaceae - chemistry; bone; Bone growth; Cell culture; Cell growth; Cell Proliferation - drug effects; Cell Shape - drug effects; Cell Survival - drug effects; Collagen; Collagen Type I - genetics; Collagen Type I - metabolism; Collagen Type I, alpha 1 Chain; Differentiation; Dose-Response Relationship, Drug; Drug dosages; Ethanol; Extracellular matrix; Fractures; Gene expression; Gene Expression Regulation - drug effects; Genes; Mice; Mineralization; Morphology; Nodules; Original Research; Osteoblastogenesis; Osteoblasts; Osteoblasts - drug effects; Osteoblasts - metabolism; Osteocalcin; Osteocalcin - genetics; Osteocalcin - metabolism; Osteogenesis; Osteogenesis - drug effects; Osteogenesis - genetics; Osteoporosis; Oxidative stress; Phosphatase; Phosphatases; Phytotherapy; Plants, Medicinal; Proteins; Seeds; Sp7 Transcription Factor - genetics; Sp7 Transcription Factor - metabolism; Staining; Stem Cells - drug effects; Stem Cells - metabolism; Time dependence; Time Factors; tocotrienol; Tocotrienols - isolation & purification; Tocotrienols - pharmacology; Vitamin E
• ISSN: 1177-8881; 1177-8881
• DOI: 10.2147/DDDT.S168935

Annatto-derived tocotrienol (AnTT) has been shown to improve bone formation in animal models of osteoporosis. However, detailed studies of the effects of AnTT on preosteoblastic cells were limited. This study was conducted to investigate the osteogenic effect of AnTT on preosteoblast MC3T3-E1 cells in a time-dependent manner. Murine MC3T3-E1 preosteoblastic cells were cultured in the different concentrations of AnTT (0.001-1 µg/mL) up to 24 days. Expression of osteoblastic differentiation markers was measured by qPCR (osterix [OSX], collagen 1 alpha 1 [COL1α1], alkaline phosphatase [ALP], and osteocalcin [OCN]) and by fluorometric assay for ALP activity. Detection of collagen and mineralized nodules was done via Direct Red staining and Alizarin Red staining, respectively. The results showed that osteoblastic differentiation-related genes, such as OSX, COL1α1, ALP, and OCN, were significantly increased in the AnTT-treated groups compared to the vehicle group in a time-dependent manner (