Effects of hemp (Cannabis sativa L. subsp. sativa) seed oil on cell viability and alkaline phosphatase activity in human osteoblast cells

Backgrounds: Osteoporosis is the most common metabolic bone disease in the elderly, especially postmenopausal women.The disease is caused by the imbalance of bone remodeling, characterized by a decrease in number and impaired functionality of osteoblasts.Hemp seed oil contains high content of polyunsaturated fatty acids and bioactive phytochemicals including cannabidiol (CBD) and polyphenols.Hemp seed oil exhibits a wide range of pharmacological properties including antioxidants and anti-inflammatory.Objectives: This study examines the effect of hemp seed oil on cell viability and osteoblast differentiation by detecting alkaline phosphatase activity in the human fetal osteoblastic cell line (hFOB1.19). Methods:The cell viability on hFOB1.19 was tested by CCK-8 assay for 24 and 48 hours at 0.25-64 g/ml concentration.Osteoblast differentiation was investigated after treatment with hemp seed oil for 5 and 7 days by both ALP enzymatic activity assay and ALP stanining using BCIP/NBT method.Results: Hemp seed oil had no cytotoxic effect with concentrations ranging from 0.5-8 g/ml for 48 hours.The cell viability significantly increased with maximally at a concentration of 4 g/ml at 48 hours.Following a treatment period of 5 and 7 days, hemp seed oil significantly increased in enzymatic ALP activity, with maximally at concentration of 8 g/ml for 5 days.Hemp seed oil also increased blue purple color of positive ALP staining for 5 days. Conclusion:Hemp seed oil promotes cell viability and osteoblast differentiation by increasing the level of alkaline phosphatase.Therefore, it might be used for promoting bone formation and prevention of osteoporosis.