Title

Effect of Vitamin C on Generation of 5-hydroxymethylcytosine in MC3T3-E1 Cells

Department

Biology

Publication Year

Spring 2015

Abstract

Osteoporosis is a serious health problem characterized by low bone mineral density and spontaneous bone fracture. Few treatments and therapies targeting this disease exist today, causing morbidity for patients and enormous healthcare costs. Thus, there is a compelling need to find novel effective therapies to promote fracture healing, or, perhaps, fracture prevention. Evidence has shown that vitamin C (ascorbic acid) is necessary for healthy bone development. Research also suggests that prolyl hydroxylase domain-containing protein 2 is necessary for vitamin C to effectively increase osteoblast differentiation. Prolyl hydroxylase domain containing protein 2 is known for its role in hydroxylating proline residues involved in collagen synthesis, but this enzyme may also hydroxylate 5-methylcytosines. Therefore the hypothesis of this study is that vitamin C plays a role in bone by inducing the hydroxylation of 5- methylcytosines through an epigenetic mechanism that could potentially lead to upregulation of genes involved in osteoblast differentiation and maturation. Treatment of pre-osteoblastic MC3T3-E1 cells with 50 μM ascorbic acid increased alkaline phosphatase activity, a biochemical marker of bone formation, more than 2-fold compared to the vehicle-control, confirming vitamin C enhancement of pre-osteoblast differentiation into osteoblasts. No significant increase in hydroxymethylation was detected upon ascorbic acid treatment through dot blot analysis or an ELISA. Additional investigation into whether vitamin C induces hydroxymethylation in MC3T3-E1 cells is needed in order to further address the role of prolyl hydroxylase domain-containing protein 2 in bone development. This information may inform drug treatment therapies for osteoporosis, thus helping to address the worldwide burden of this disease.

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