Abstract
Although low-intensity pulsed ultrasound (LIPUS) has been shown to enhance bone fracture healing, the underlying mechanism of LIPUS remains to be fully elucidated. Here, to better understand the molecular mechanism underlying cellular responses to LIPUS, we investigated gene expression profiles in mouse MC3T3-E1 preosteoblast cells exposed to LIPUS using high-density oligonucleotide microarrays and computational gene expression analysis tools. Although treatment of the cells with a single 20-min LIPUS (1.5 MHz, 30 mW/cm2) did not affect the cell growth or alkaline phosphatase activity, the treatment significantly increased the mRNA level of Bglap. Microarray analysis demonstrated that 38 genes were upregulated and 37 genes were downregulated by 1.5-fold or more in the cells at 24-h post-treatment. Ingenuity pathway analysis demonstrated that the gene network U (up) contained many upregulated genes that were mainly associated with bone morphology in the category of biological functions of skeletal and muscular system development and function. Moreover, the biological function of the gene network D (down), which contained downregulated genes, was associated with gene expression, the cell cycle and connective tissue development and function. These results should help to further clarify the molecular basis of the mechanisms of the LIPUS response in osteoblast cells.
Highlights
In many medical fields, ultrasound (US) has proven to be an effective diagnostic or therapeutic tool [1,2]
We investigated the changes in gene expression in MC3T3-E1 preosteoblast cells treated with low-intensity pulsed ultrasound (LIPUS) by using a GeneChip® microarray analysis system in order to better understand the molecular mechanisms underlying cellular responses to this stress
When MC3T3-E1 cells were exposed to LIPUS (30 mW/cm2, for 20 min), followed by culturing at 37 °C for 0 to 48 h, the cell number was gradually increased in a time-dependent fashion
Summary
Ultrasound (US) has proven to be an effective diagnostic or therapeutic tool [1,2]. With the help of novel transcript profiling technology, a view of the genome-wide expression profiles can be assayed simultaneously, allowing scanning differential expression of a large number of genes This technology has been used to analyze the expression of genes in response to LIPUS in human osteoblastic osteosarcoma MG-63 [28] and SaOS-2 cells [29]. We have used pathway analysis technologies to map gene expression data into relevant gene networks on the basis of their functional annotation and known molecular interactions [39,40,41] Using these technologies, unique gene networks that are associated with cellular development and cell death were identified in human lymphoma U937 cells treated with LIPUS [39]. We investigated the changes in gene expression in MC3T3-E1 preosteoblast cells treated with LIPUS by using a GeneChip® microarray analysis system in order to better understand the molecular mechanisms underlying cellular responses to this stress
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