Effects of CDP-choline on osteogenic differentiation of pre-osteoblastic MC3T3-E1 cells

Abstract

The cholinergic system, traditionally associated with neurotransmission, has been recognized for its involvement in bone physiology and osteogenesis. This study investigates the effects of CDP-choline, a compound known for its neuroprotective properties, on the osteogenic differentiation of pre-osteoblastic MC3T3-E1 cells. The objective was to determine if CDP-choline could enhance key markers of osteogenesis, such as alkaline phosphatase (ALP) activity, hydroxyproline (HYP) content, intracellular calcium levels, and collagen production. Mouse pre-osteoblast MC3T3-E1 cells were cultured and differentiated in osteogenic media supplemented with CDP-choline. Cell viability was assessed using the MTT assay. Intracellular calcium levels were measured using a spectrofluorometric assay. ALP and HYP levels were determined using ELISA. Statistical significance was analysed using ANOVA and Student’s t-test. CDP-choline treatment significantly enhanced ALP activity in both cell lysates and media, indicating early osteogenic differentiation. HYP levels were also significantly elevated, suggesting enhanced collagen synthesis and bone matrix stability. Intracellular calcium levels increased, confirming active osteogenic differentiation potential of CDP-choline. The findings demonstrate that CDP-choline significantly promotes osteogenic differentiation in pre-osteoblastic MC3T3-E1 cells. The increase in ALP activity, HYP content, intracellular calcium levels highlight its potential as a natural agent for bone health and regeneration therapies. These results suggest that CDP-choline could be a promising candidate for developing new treatments for bone-related disorders.