In vitro osteogenic activity of the Malaysian Kelulut Honey on murine preosteoblasts MC3T3-E1 cells

Malaysian Kelulut Honey (MKH) is a well-recognised Malaysian honey and has gained widespread popularity due to its ability to promote health. Numerous scientific studies have been done to prove the health benefits of MKH; however, there is no evidence on the bone protective effects of MKH in osteogenic condition studied yet. Thus, this study aimed to explore the effect of MKH on bone by using murine preosteoblast cell line (MC3T3-E1) with an intention to establish MKH as one of the natural products which may promote osteoblast formation. The MC3T3-E1 were cultured and treated with honey at various concentration (0.001% v/v to 10.0% v/v) in a 96-well plate and incubated at 37°C with 5% CO2. Cytotoxicity test of MKH on the viability and proliferation of MC3T3- E1 cells were assessed by using 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay and bromodeoxyuridine (BrdU) assay up to 72 hours. The concentrations of MKH in the range of 0.01% v/v to 0.1% v/v showed no cytotoxic effect on MC3T3-E1 cells. Besides cell growth, the ability of MKH to promote osteoblastic differentiation was also investigated. The αMEM supplemented with 100 nM dexamethasone, 0.05 μM ascorbate-2-phosphate, and 10 mM β-glycerophosphate was used as negative control. The result of osteoblastogenesis showed that ALP activity (p<0.05) and deposition of collagen (p<0.001) increased significantly when compared to their negative control respectively on day 21. However, there were increased in other aspects of osteoblastic differentiation, such as colony formation, calcium and inorganic- phosphate deposition, concentration of osteocalcin (OCN) and osteoprotegerin (OPG) and bone-specific marker mRNAs expressions on day 21 without any significant difference compared to the negative control. Hence, the role of MKH on osteoblasts in osteogenic activity requires further investigation.

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