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Effect of Amelogenin Coating of a Nano-Modified Titanium Surface on Bioactivity

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Effect of Amelogenin Coating of a Nano-Modified Titanium Surface on Bioactivity

Chisato Terada et al. Int J Mol Sci.

Abstract

The interactions between implants and host tissues depend on several factors. In particular, a growing body of evidence has demonstrated that the surface texture of an implant influences the response of the surrounding cells. The purpose of this study is to develop new implant materials aiming at the regeneration of periodontal tissues as well as hard tissues by coating nano-modified titanium with amelogenin, which is one of the main proteins contained in Emdogain®. We confirmed by quartz crystal microbalance evaluation that amelogenin is easy to adsorb onto the nano-modified titanium surface as a coating. Scanning electron microscopy, scanning probe microscopy, X-ray photoelectron spectroscopy, and Fourier-transform infrared spectroscopy analyses confirmed that amelogenin coated the nano-modified titanium surface following alkali-treatment. In vitro evaluation using rat bone marrow and periodontal ligament cells revealed that the initial adhesion of both cell types and the induction of hard tissue differentiation such as cementum were improved by amelogenin coating. Additionally, the formation of new bone in implanted surrounding tissues was observed in in vivo evaluation using rat femurs. Together, these results suggest that this material may serve as a new implant material with the potential to play a major role in the advancement of clinical dentistry.

Keywords: amelogenin; host-implant interaction; nanostructure; tissue regeneration.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
SEM micrographs of control and test groups.
Figure 2
Figure 2
Scanning probe (SP) micrographs of control and test groups.
Figure 3
Figure 3
Wide scan of the test and control groups.
Figure 4
Figure 4
FTIR analysis of test and control groups.
Figure 5
Figure 5
Quartz crystal microbalance (QCM) analysis of test and control groups. * p < 0.05.
Figure 6
Figure 6
Morphology of rat bone marrow (RBM) cells and rat periodontal ligament cells in the control and test groups after culturing for 24 h. Actin filaments (green) were labeled with Alexa Fluor 488-phalloidin and nuclei (blue) were stained with 4′,6-diamidino-2-phenylindole (DAPI).
Figure 7
Figure 7
Cell adhesion in control and test groups. * p < 0.05.
Figure 8
Figure 8
Quantitative real-time (qRT)-PCR analysis of osteogenesis and cementum related gene expression in control and test groups. * p < 0.05.
Figure 9
Figure 9
ALP activity in control and test groups. * p < 0.05.
Figure 10
Figure 10
OCN production in control and test groups. * p < 0.05.
Figure 11
Figure 11
Ca deposition in test and control groups. * p < 0.05.
Figure 12
Figure 12
Micro-CT images (The implants were marked with red color, cortical bone with blue color, and cancellous bone with green color).
Figure 13
Figure 13
Quantitative evaluation of the trabecular bone within ROI (BV/TV, Tb.N, Tb.Th and Tb. Sp). * p < 0.05.
Figure 14
Figure 14
The longitudinally undecalcified histological sections with implants and peri-implant bones. (Purple color: osteoid, white color: new bone).
Figure 15
Figure 15
Quantitative histomorphometric analysis within the region of measurement above (BIC and BA). * p < 0.05.
Figure 16
Figure 16
The histological sections were also observed using confocal laser scanning microscopy for dynamic histomorphometry according to fluorescence labeling. (Blue color: 1 week of new bone, yellow color: 4 week of new bone, green color: 8 week of new bone, red color: osteoid).
Figure 17
Figure 17
The percentage of labeled bone area (%LBA). * p < 0.05.

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