The Evaluation of Osteoblastic Cell Behavior on Treated Titanium Surface

Maria A. S. De Souza Alencar1, Elizabeth F. Martinez1, Fábio C. Figueiredo1, André R. De Lima e Silva1, José E. Protazio1, Maicon Bertamoni1, Daiane C. Peruzzo1, Marcelo H. Napimoga1, *
1 Laboratory of Immunology and Molecular Biology, São Leopoldo Mandic Institute and Research Center, Campinas, SP, Brazil

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© 2020 Alencar et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: ( This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Laboratory of Immunology and Molecular Biology, São Leopoldo Mandic Institute and Research Center, R. José Rocha Junqueira, 13 - 13045-755-Campinas, SP, Brazil; Tel: +55 19 3211-3627; Fax: +55 19 3211-3712; E-mails: or



There are several potential advantages in optimizing the initial events of osseointegration in the benefit of clinical outcome.


The objective of the present study was to evaluate the behavior of osteoblastic cells on surfaces treated by double acid etching using HNO3 and H2SO4.


Commercially pure titanium (grade 4) discs measuring 6 mm in diameter and 2 mm in thickness were used. The discs were divided into two groups: machined group and double acid-etched discs (HNO3 and H2SO4). Surface characteristics were assessed using Scanning Electron Microscopy. Pre-osteoblastic MC3T3-E1 cells were used for cell culture on the tested surfaces to assess proliferation, viability (MTT), as well as secretion (ELISA) and cytoplasmic expression (Western blot) of type I collagen.


The data obtained were analyzed using t-test or two-way ANOVA followed by Bonferroni’s test at 95% significance. The titanium surfaces showed average roughness values for the machined and treated surfaces of 0.29 and 1.16, respectively (p<0.05). An increase in cell proliferation was observed, which was corroborated by the viability assay. Both type I collagen secretion and intracellular expression were higher on the double acid-etched surface compared to the machine surfaces (p<0.05).


Implant surfaces treated by double acid etching positively affected the early events of the interaction between titanium and osteoblastic cells, suggesting optimization of osseintegration.

Keywords: Dental implant, Osseointegration, Surface roughness, Osteoblastic cell, Titanium, HNO3 and H2SO4.