Semi-quantitative RT-PCR analysis of LIM mineralization protein 1 and its associated molecules in cultured human dental pulp cells

Objective: LIM mineralization protein 1 (LMP-1), an intracellular signaling molecule, regulates osteoblast differentiation and maturation, as well as bone formation. However, the role of LMP-1 in the differentiation of human dental pulp cells and formation of dentin has not been determined. The study was to investigate the expression of LMP-1, the related proteins, such as bone morphogenetic proteins 2, 6 and 7 (BMP-2, BMP-6 and BMP-7), and core binding factor alpha 1 (Cbfa1) during the differentiation of cultured human dental pulp cells and the formation of mineralized nodules. Design: Differentiation of human dental pulp cells was induced by dexamethasone, asorbic acid and p-glycerophosphate. The formation of mineralized nodules, was determined by Von Kossa staining and immunocytochemistry detection of dentin sialoprotein. Expression of LMP-1, the related proteins and the differentiation marker alkaline phosphatase (ALP) was analysed by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: The expression of LMP-1, BMP-2, BMP-6, BMP-7 and Cbfa1 was significantly increased in the process of dental pulp cells differentiation and the formation of mineralized nodules, while the pattern of the expression was distinct. Conclusions: The elevated level of LMP-1, BMPs and Cbfa1 expression indicated they might play a role in the differentiation of human dental pulp cells and the formation of mineralized nodules. (C) 2007 Elsevier Ltd. All rights reserved.