Inhibitory effects of ameloblastin on epithelial cell proliferation

Saito, Noriko; Ariyoshi, Wataru; Okinaga, Toshinori; Kamegawa, Mariko; Matsukizono, Miho; Akebiyama, Yasuo; Kitamura, Chiaki; Nishihara, Tatsuji

doi:10.1016/j.archoralbio.2014.05.010

Cited by 8 publications

(6 citation statements)

References 28 publications

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“…Our findings are strongly supported by the previous studies in which the proliferation of SCC-25 cells was shown to be inhibited via cell cycle arrest at G1 phase in response to EMD 16 or ameloblastin 23 . Furthermore, the binding of enamel matrix proteins to specific proteins including fibronectin and type I collagen regulates their adhesive properties, leading to modulate fibroblast adhesion to epithelial cells 19 .…”

Section: Discussionsupporting

confidence: 89%

“…Our results suggest that ameloblastin is the principal bioactive factor in EMD in inhibiting epithelial cell proliferation. Recent study revealed that lower a concentration (12.5-100 ng/ml) of recombinant human ameloblastin purified using HaloTag ® fusion tag system suppressed the proliferation of SCC-25 cells 23 . We have no ready explanation for these discrepancies, though it is possible that they reflect differences in the purification procedure of recombinant protein.…”

Section: Discussionmentioning

confidence: 83%

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Novel biological activity of ameloblastin in enamel matrix derivative

et al. 2015

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Objective Enamel matrix derivative (EMD) is used clinically to promote periodontal tissue regeneration. However, the effects of EMD on gingival epithelial cells during regeneration of periodontal tissues are unclear. In this in vitro study, we purified ameloblastin from EMD and investigated its biological effects on epithelial cells.Material and Methods Bioactive fractions were purified from EMD by reversed-phase high-performance liquid chromatography using hydrophobic support with a C18 column. The mouse gingival epithelial cell line GE-1 and human oral squamous cell carcinoma line SCC-25 were treated with purified EMD fraction, and cell survival was assessed with a WST-1 assay. To identify the proteins in bioactive fractions of EMD, we used proteome analysis with two-dimensional gel electrophoresis followed by identification with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis.Results Purified fractions from EMD suppressed proliferation of GE-1 and SCC-25. LC-MS/MS revealed that ameloblastin in EMD is the component responsible for inhibiting epithelial cell proliferation. The inhibitory effect of ameloblastin on the proliferation of GE-1 and SCC-25 was confirmed using recombinant protein.Conclusion The inhibitory effects of EMD on epithelial cell proliferation are caused by the biological activities of ameloblastin, which suggests that ameloblastin is involved in regulating epithelial downgrowth in periodontal tissues.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 83%

Novel biological activity of ameloblastin in enamel matrix derivative

et al. 2015

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“…This result confirms a significant role for the Lam peptide in promoting keratinocyte growth 35 , but contradicts previous reports on the effects of Ambn peptide on downregulation of epithelial proliferation. 36 Moreover, the peptide coatings stimulated HDs formation by keratinocytes. Our hypothesis was validated as coatings with both Lam and Ambn peptides synergistically enhanced HDs formation with AMBN+LAM surfaces inducing significantly higher HDs formation than all monopeptide-coated and control surfaces.…”

Section: Discussionmentioning

confidence: 99%

Peptide coatings enhance keratinocyte attachment towards improving the peri-implant mucosal seal

et al. 2018

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There is a critical need for preventing peri-implantitis as its prevalence has increased and dental implants lack features to prevent it. Research strategies to prevent peri-implantitis have focused on modifying dental implants to incorporate different antimicrobial agents. An alternative strategy consists of barring the expansion of the biofilm subgingivally by forming a long-lasting permucosal seal between the soft tissue and the implant surface. Here, we innovatively biofunctionalized titanium with bioinspired peptide coatings to strengthen biological interactions between epithelial cells and the titanium surface. We selected laminin 332- and ameloblastin-derived peptides (Lam, Ambn). Laminin 332 participates in the formation of hemidesmosomes by keratinocytes and promotes epithelial attachment around teeth; and ameloblastin, an enamel derived protein, is involved in tissue regeneration events following disruption of the periodontium. Lam, Ambn or combinations of both peptides were covalently immobilized on titanium discs. Successful immobilization of the peptides was confirmed by contact angle goniometry, X-ray photoelectron spectroscopy and fluorescent labelling of the peptides. Additionally, we confirmed the mechanical and thermochemical stability of the peptides on Ti substrates. Proliferation and hemidesmosome formation of human keratinocytes (TERT-2/OKF-6) were assessed by immunofluorescence labelling. The peptide-coated surfaces increased cell proliferation for up to 48 h in culture compared to control surfaces. Most importantly, formation of hemidesmosomes by keratinocytes was significantly increased on surfaces coated with Ambn + Lam peptides compared to control (p < 0.01) and monopeptide coatings (p < 0.005). Together, these results support the Ambn + Lam multipeptide coating as a promising candidate for inducing a permucosal seal around dental implants.

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“… 26 For example, Saito et al isolated recombinant ameloblastin from COS-7 cells to study the inhibitory effects of ameloblastin on epithelial cell proliferation. 27 In this study, HaloTag was utilized for both the isolation and cellular tracking of ameloblastin, allowing researchers to show that ameloblastin induced cell cycle arrest in epithelial cells that led to periodontitis.…”

Section: Protein Isolation and Purificationmentioning

confidence: 99%

HaloTag Technology: A Versatile Platform for Biomedical Applications

2015

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Exploration of protein function and interaction is critical for discovering links between genomics, proteomics, and disease state; yet the immense complexity of proteomics found in biological systems currently limit our investigational capacities. While affinity and autofluorescent tags are widely employed for protein analysis, these methods have limited success as they lack specificity and require multiple fusion tags and genetic constructs. As an alternative approach, the innovative HaloTag protein fusion platform allows for comprehensive analysis of protein function and interaction using a single genetic construct with multiple capabilities. This is accomplished using a simplified process, in which a variable HaloTag ligand binds rapidly to the HaloTag protein (usually linked to the protein of interest) with high affinity and specificity. In this review article, we examine all current applications of the HaloTag technology platform for biomedical applications such as the study of protein isolation and purification, protein function, protein-protein and protein-DNA interactions, biological assays, in vitro cellular imaging, and in vivo molecular imaging. In addition, novel uses of the HaloTag platform are briefly discussed with potential future applications.

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Inhibitory effects of ameloblastin on epithelial cell proliferation

Cited by 8 publications

References 28 publications

Novel biological activity of ameloblastin in enamel matrix derivative

Novel biological activity of ameloblastin in enamel matrix derivative

Peptide coatings enhance keratinocyte attachment towards improving the peri-implant mucosal seal

HaloTag Technology: A Versatile Platform for Biomedical Applications

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