Induction of enamel matrix protein expression in an ameloblast cell line co-cultured with a mesenchymal cell line in vitro

Matsumoto, Asako; Harada, Hidemitsu; Saito, Masahiro; Taniguchi, Akiyoshi

doi:10.1007/s11626-010-9362-7

Cited by 23 publications

(23 citation statements)

References 26 publications

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“…The HAT-7 cell line is an immortalized dental epithelial cell line derived from the cervical stem cell population of rat mandibular incisors. It can shift from the transient amplification stage to ameloblast lineage cells (26). SDC4 expression was downregulated by siRNA, which had no effect on the gene expression of SDC1, 2 or 3, the other three known members of the SDC family (27).…”

Section: Discussionmentioning

confidence: 95%

Expression and roles of syndecan-4 in dental epithelial cell differentiation

Zhang

Yang

Sun

et al. 2014

International Journal of Molecular Medicine

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Abstract. Syndecan-4 (SDC4), a transmembrane heparan sulfate proteoglycan, acts as a signal transducer. It affects the growth and differentiation of a number of tissues and organs. However, the specific mechanisms through which SDC4 regulates the differentiation of dental epithelial cells (amelogenesis) and tooth development remains largely unknown. In the present study, to identify the SDC4-regulated processes in dental epithelial cells, the SDC4 expression pattern was examined in mouse molar and postnatal incisor tooth germs during the late bell stage of development. Small interfering RNA (siRNA) was designed for this study and used to downregulate SDC4 expression in the rat dental epithelial cell line, HAT-7. The results revealed that SDC4 was mainly present in the oral epithelium, the dental epithelial cells of enamel organs in the molars and the cervical loops in the incisors. When the inner enamel epithelial cells gave rise to ameloblasts, however, the loss of SDC4 expression was evident. SDC4 was also expressed in stratum intermedium (SI) cells in the incisors and in dental mesenchymal cells adjacent to the cervical loops in molars (E18) and postnatal incisors. Fibroblast growth factor 10 (FGF10) promoted proliferation and slightly decreased cell differentiation. The knockdown of SDC4 using specific siRNA led to a decrease in cell proliferation and a highly significant increase in amelogenin, ameloblastin, kallikrein 4 and matrix metalloproteinase 20 expression, molecules that are known to participate in the formation of enamel. These effects were attenuated by FGF10, which upregulated SDC4 expression. Taken together, these results suggest that SDC4 participates in amelogenesis, and FGF10 may modulate dental epithelial cell behaviors through the regulation of SDC4 expression. IntroductionSyndecan (SDC)4 is a cell surface heparan sulfate proteoglycan (HSPG). Its heparan sulfate (HS) chains and core protein could control the stability, movement and reception of diffusible heparin-binding growth factors (1). In addition, it can form physical connections between the extracellular matrix (ECM) and intracellular signaling to affect the growth and differentiation of a number of tissues and organs (2,3). SDC4 is highly complex by virtue of its external side chains, and thus interacts with a variety of ligands, such as vascular endothelial growth factors (VEGFs), platelet-derived growth factors (PDGFs) and fibroblast growth factors (FGFs) (4). At the cell membrane, SDC4 stabilizes the interactions between ligands and receptors by forming a ternary complex, which has been demonstrated in several signaling pathways (5,6). Proteolytic cleavage and shedding of its extracellular domain can spread FGF signaling to adjacent cells, thus regulating the local reception of FGF signaling activity (7). Its cytoplasmic domains can also initiate FGF-induced signaling independently of FGFRs through the activation of Rho GTPases, such as Rac1, which plays an essential role in the dental epithelium, involving cell-matrix interactio...

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Section: Discussionmentioning

confidence: 95%

Expression and roles of syndecan-4 in dental epithelial cell differentiation

Zhang

Yang

Sun

et al. 2014

International Journal of Molecular Medicine

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“…The reaction mixture was composed of 10 μL of SYBR Premix Ex Taq II (Takara), 10 pmol each of the forward and reverse primers, 2 μL of cDNA, and distilled water to a final volume of 20 μL. The thermocycling conditions were 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s and 60 °C for 34 s. Normalization of the data was performed using the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous control in the same reaction as the gene of interest [49]. The primers used in this study were as follows: for GAPDH, forward primer 5′-CCCCCACCACACTGAATCTC-3′ and reverse primer 5′-GCCCCTCCCCTCTTCAAG-3′; for heme oxygenase 1 (HMOX-1), forward primer 5′-GGGTGATAGAAGAGGCCAAGA-3′ and reverse primer 5′-AGCTCCTGCAACTCCTCAAA-3′; for IL-6, forward primer 5′-TGAGTACAAAAGTCCTGA-3′ and reverse primer 5′-TCTGTGCCT GCAGCTTCGT-3′; for granulocyte-macrophage colony stimulating factor 2 (CSF-2), forward primer 5′-TCTCAGAAATGTTTGACCTCCA-3′ and reverse primer 5′-GCCCTTGAGC TTGGTGAG-3′.…”

Section: Methodsmentioning

confidence: 99%

Effect of Polyethylene Glycol Modification of TiO2 Nanoparticles on Cytotoxicity and Gene Expressions in Human Cell Lines

Mano

Kanehira

Sonezaki

et al. 2012

IJMS

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Nanoparticles (NPs) are tiny materials used in a wide range of industrial and medical applications. Titanium dioxide (TiO2) is a type of nanoparticle that is widely used in paints, pigments, and cosmetics; however, little is known about the impact of TiO2 on human health and the environment. Therefore, considerable research has focused on characterizing the potential toxicity of nanoparticles such as TiO2 and on understanding the mechanism of TiO2 NP-induced nanotoxicity through the evaluation of biomarkers. Uncoated TiO2 NPs tend to aggregate in aqueous media, and these aggregates decrease cell viability and induce expression of stress-related genes, such as those encoding interleukin-6 (IL-6) and heat shock protein 70B’ (HSP70B’), indicating that TiO2 NPs induce inflammatory and heat shock responses. In order to reduce their toxicity, we conjugated TiO2 NPs with polyethylene glycol (PEG) to eliminate aggregation. Our findings indicate that modifying TiO2 NPs with PEG reduces their cytotoxicity and reduces the induction of stress-related genes. Our results also suggest that TiO2 NP-induced effects on cytotoxicity and gene expression vary depending upon the cell type and surface modification.

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“…We previously found that ameloblastin expression in HAT-7 cells was induced by co-culture with BCPb8 cells by using the collagen membrane (Matsumoto et al 2011). We have constructed two co-culture models to investigate the interaction between two different dental cell lines, a rat HAT-7 cell line that originated from dental epithelia, and bovine BCPb8 cells, derived from cementoblast progenitor cells.…”

Section: Resultsmentioning

confidence: 99%

“…During tooth development, various growth factors, such as, fibroblast growth factor, transforming growth factor-β, and insulin-like growth factor (IGF), regulate cell proliferation and differentiation in a paracrine or autocrine fashion (review by Joseph et al 1996; Thesleff and Mikkola 2002; Thesleff 2003). To investigate the interaction between epithelial and mesenchymal cells, we previously constructed a three-dimensional culture system by using a collagen membrane, which induces the differentiation of epithelial cells (Matsumoto et al 2011). Our results suggest that soluble factors produced by mesenchymal cells contribute to the induction of epithelial cell differentiation.…”

Section: Introductionmentioning

confidence: 99%

Induction of insulin-like growth factor 2 expression in a mesenchymal cell line co-cultured with an ameloblast cell line

Matsumoto

Harada

Saito

et al. 2011

In Vitro Cell.Dev.Biol.-Animal

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Various growth factors have been implicated in the regulation of cell proliferation and differentiation during tooth development. It has been unclear if insulin-like growth factors (IGFs) participate in the epithelium–mesenchyme interactions of tooth development. We previously produced three-dimensional sandwich co-culture systems (SW) containing a collagen membrane that induce the differentiation of epithelial cells. In the present study, we used the SW system to analyze the expression of IGFs and IGFRs. We demonstrate that IGF2 expression in mesenchymal cells was increased by SW. IGF1R transduces a signal; however, IGF2R does not transduce a signal. Recombinant IGF2 induces IGF1R and IGF2R expression in epithelial cells. IGF1R expression is increased by SW; however, IGF2R expression did not increase by SW. Thus, IGF2 signaling works effectively in SW. These results suggest that IGF signaling acts through the collagen membrane on the interaction between the epithelium and mesenchyme. In SW, other cytokines may be suppressed to induce IGF2R induction. Our results suggest that IGF2 may play a role in tooth differentiation.

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Induction of enamel matrix protein expression in an ameloblast cell line co-cultured with a mesenchymal cell line in vitro

Cited by 23 publications

References 26 publications

Expression and roles of syndecan-4 in dental epithelial cell differentiation

Expression and roles of syndecan-4 in dental epithelial cell differentiation

Effect of Polyethylene Glycol Modification of TiO2 Nanoparticles on Cytotoxicity and Gene Expressions in Human Cell Lines

Induction of insulin-like growth factor 2 expression in a mesenchymal cell line co-cultured with an ameloblast cell line

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