Abstract

The aim of this work was to observe the behavior of osteoblast cells cultured in vitro on titanium discs in relation to disc surface roughness and the addition of melatonin to the culture medium. MG63 osteoblast cells were cultivated on 120 Grade 5 Ti divided into three groups: Group E, treated with dual acid etch; Group EP, treated with dual acid etch and calcium phosphate; and Group M, machined. Surface roughness was examined under a laser scanning confocal microscope (CLSM) and scanning electron microscopy (SEM). The proliferation and morphology of cells were determined under fluorescence microscopy and SEM. Messenger ribonucleic acid (mRNA) of different genes related to osteoblastic differentiation was quantified by means of real-time quantitative polymerase chain reaction (RT-PCR) assay. The greatest surface roughness was found in Group EP (Ra 0.354 µm), followed by Group E (Ra 0.266 µm), and Group M (Ra 0.131 µm), with statistically significant differences between the groups (p < 0.001). In the presence of melatonin a trend to a higher cell proliferation was observed in all groups although significant differences were only found in Group M (p = 0.0079). Among the genes studied, a significant increase in phosphate-regulating neutral endopeptidase, X-linked (PHEX) expression was observed in cells cultured on EP discs. The addition of melatonin increased osteoblast cell proliferation and differentiation, and may favor the osseointegration of dental implants.

Highlights

  • The clinical success of dental implants depends on osseointegration, the direct structural and functional connection between living bone and the implant surface [1]

  • To analyze the influence of the addition of melatonin to the cell culture media on the adhesion, proliferation, morphology, and differentiation of human osteoblast cells cultured on Grade 5 Ti with different surface treatments

  • Quantification of Specific Messenger ribonucleic acid (mRNA) Quantification of relative expression of different genes related to osteoblast differentiation was perfoQrmuaendtibfiycacatilocnuloatfinreglathtievevaelxuperoesf s2i−oΔnΔCot, fudsiinffgertehnetegnednoegserneoluatseedxtporeossstieoonbolafstthde igffeenreenGtAiaPtiDonHwasasa performed by calculating the value of 2−ΔΔCt, using the endogenous expression of the gene Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a

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Summary

Introduction

The clinical success of dental implants depends on osseointegration, the direct structural and functional connection between living bone and the implant surface [1]. To study the surface topography and roughness (Ra) of machined Grade 5 Ti, treated with dual acid etching or dual acid etching plus calcium phosphate coating, prior to cell culture; 2. To analyze the influence of the addition of melatonin to the cell culture media on the adhesion, proliferation, morphology, and differentiation of human osteoblast cells cultured on Grade 5 Ti with different surface treatments. Ed in the three study groups: M (machined), E (dual acid etched) and EP (dual acid etched and calcium phosphate coating) according to culture time and thTeaabdled2i.tCioelnl doefnsmityeolaf tcoelnlsicnulttourtehdeoncutitlatnuiuremmdiescds.iuDmesc.riptive results of mean cell density ± SD by experimental group (discs M, E, and EP), cell culture time (24 and 72 h), and without or with the. Quantification of Specific mRNA Quantification of relative expression of different genes related to osteoblast differentiation was perfoQrmuaendtibfiycacatilocnuloatfinreglathtievevaelxuperoesf s2i−oΔnΔCot, fudsiinffgertehnetegnednoegserneoluatseedxtporeossstieoonbolafstthde igffeenreenGtAiaPtiDonHwasasa performed by calculating the value of 2−ΔΔCt, using the endogenous expression of the gene GAPDH as a

Quantification of Specific mRNA
Materials and Methods
Cell Culture
Findings
Statistical Analysis
Full Text
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