Abstract
BackgroundEpithelial invagination is important for initiation of ectodermal organogenesis. Although many factors regulate ectodermal organogenesis, there is not any report about their functions in real-time study. Electric cell-substrate impedance sensing (ECIS), a non-invasive, real-time surveillance system, had been used to detect changes in organ cell layer thickness through quantitative monitoring of the impedance of a cell-to-microelectrode interface over time. It was shown to be a good method for identifying significant real-time changes of cells. The purpose of this study is to establish a combined bioengineered organ-ECIS model for investigating the real time effects of fibroblast growth factor-9 (FGF-9) on epithelial invagination in bioengineered ectodermal organs. We dissected epithelial and mesenchymal cells from stage E14.5 murine molar tooth germs and identified the real-time effects of FGF-9 on epithelial-mesenchymal interactions using this combined bioengineered organ-ECIS model.ResultsMeasurement of bioengineered ectodermal organ thickness showed that Fibroblast growth factor-9 (FGF-9) accelerates epithelial invagination in reaggregated mesenchymal cell layer within 3 days. Gene expression analysis revealed that FGF-9 stimulates and sustains early Ameloblastin and Amelogenin expression during odontogenesis.ConclusionsThis is the first real-time study to show that, FGF-9 plays an important role in epithelial invagination and initiates ectodermal organogenesis. Based on these findings, we suggest FGF-9 can be applied for further study in ectodermal organ regeneration, and we also proposed that the ‘FGF-BMP balancing system’ is important for manipulating the morphogenesis of ectodermal organs. The combined bioengineered organ-ECIS model is a promising method for ectodermal organ engineering and regeneration research.
Highlights
Epithelial invagination is important for initiation of ectodermal organogenesis
Studies have shown that Fibroblast growth factor (FGF)-8 and fibroblast growth factor-9 (FGF-9) are expressed within the murine molar epithelium: FGF-8 has a specific role in multicuspid odontogenesis [13,14]; in this study, we investigated the role of FGF-9, present within the oral epithelium of the first branchial arch at stage E10
Preliminary results showed that FGF-9 can induce early, significant Ameloblastin, Amelogenin and Osteocalcin expression in mesenchymal cells cultured with FGF-9 (Additional file 1)
Summary
Many factors regulate ectodermal organogenesis, there is not any report about their functions in real-time study. The purpose of this study is to establish a combined bioengineered organ-ECIS model for investigating the real time effects of fibroblast growth factor-9 (FGF-9) on epithelial invagination in bioengineered ectodermal organs. We dissected epithelial and mesenchymal cells from stage E14.5 murine molar tooth germs and identified the real-time effects of FGF-9 on epithelial-mesenchymal interactions using this combined bioengineered organ-ECIS model. Many factors regulate ectodermal organogenesis, observing their functions in real-time is difficult. The multipotency of embryonic stem cells makes creation of a non-invasive, noninductive environment for the observation of specific growth factor functions within ectodermal organ development challenging. Ectodermal organs are greatly affected by their in vivo environment and its inherent regulating factors
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