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Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XVIII | Pages 29 - 29
1 May 2012
Zou H Brookes S Lyngstadaas S Boccaccini A Tsiridis E Seedhom B Yang X
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Objective

Human bone marrow stromal cells (HBMSCs) are multipotent and can form bone, cartilage or other tissues under different inductive conditions. The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on the growth and osteogenic differentiation of HBMSCs.

Methods

HBMSCs were cultured in monolayer with EMD (1, 10, 50,100, 250μg/ml) in aMEM supplemented with 2% FBS for 3 days. Cells cultured in aMEM supplemented with 2% FBS (basal medium) served as the control group. Double-stranded DNA was quantified by PicoGreen assay. Quantitative RT-PCR was performed to determine the expression levels of RUNX2, osteopontin (OPN) and osteocalcin (OCN), dentin matrix protein1 (DMP1) and dentin sialophosphoprotein (DSPP) at different time points (day 0, 5 and 10) when exposed to 10μg/ml EMD or basal medium. Alkaline phosphatase specific activity (ALPSA) was determined after 5 and 10 days culture. Mineral deposition (as calcium) was visualised using Alizarin Red staining.