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

Abstract

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 µg/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 µg/ml for 48 hours. The cell viability significantly increased with maximally at a concentration of 4 µg/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 µg/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.