Abstract

Beta-titanium alloys are promising materials for bone implants due to their advantageous mechanical properties. For enhancing the interaction of bone cells with this perspective material, we developed a ferroelectric barium titanate (BaTiO3) coating on a Ti39Nb alloy by hydrothermal synthesis. This coating was analyzed by scanning electron and Raman microscopy, X-ray diffraction, piezoresponse force microscopy, X-ray photoelectron spectroscopy, nanoindentation, and roughness measurement. Leaching experiments in a saline solution revealed that Ba is released from the coating. A progressive decrease of Ba concentration in the material was also found after 1, 3, and 7 days of cultivation of human osteoblast-like Saos-2 cells. On day 1, the Saos-2 cells adhered on the BaTiO3 film in higher initial numbers than on the bare alloy, but they were less spread, and their initial proliferation rate was slower. These cells also contained a lower amount of beta1-integrins and vinculin, i.e., molecules involved in cell adhesion, and produced a lower amount of collagen I. This cell behavior was attributed to a higher surface roughness of BaTiO3 film rather than to its potential cytotoxicity, because the cell viability on this film was very high, reaching almost 99%. The amount of alkaline phosphatase, an enzyme involved in bone matrix mineralization, was similar in cells on the BaTiO3-coated and uncoated alloy, and on day 7, the cells on BaTiO3 film attained a higher final cell population density. These results indicate that after some improvements, particularly in its roughness and stability, the hydrothermal ferroelectric BaTiO3 film could be promising coating for improved osseointegration of bone implants.

Highlights

  • Bone implants are of increasing importance in orthopedic, dental, plastic, and reconstructive surgery for treating or replacing hard tissues damaged by various diseases or by trauma

  • For enhancing the interaction of bone cells with this perspective material, we developed a ferroelectric barium titanate (BaTiO3) coating on a Ti39Nb alloy by hydrothermal synthesis

  • These results indicate that after some improvements, in its roughness and stability, the hydrothermal ferroelectric BaTiO3 film could be promising coating for improved osseointegration of bone implants

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Summary

Introduction

Bone implants are of increasing importance in orthopedic, dental, plastic, and reconstructive surgery for treating or replacing hard tissues damaged by various diseases (osteoarthrosis, osteoarthritis, tumors, inborn malformations) or by trauma (e.g., traffic accidents and industrial accidents, sports injuries etc.). Other methods for growing a BaTiO3 film with ferroelectric properties on titanium implants are electrostatic spray pyrolysis (ESP) [36], which combines the biocompatibility of a ceramic with the high mechanical strength of a metal, and methods based on electrochemical principles, such as micro-arc oxidation [37]. Another promising method, which we intended to explore in this study, is the hydrothermal method. The presence and amount of selected specific markers of cell adhesion, namely β1-integrins and vinculin, and early markers of osteogenic cell differentiation, namely ALP and collagen I, were evaluated in Saos-2 cells by measuring the intensity of fluorescence after immunofluorescence staining of these molecules

Preparation of Substrates for Ferroelectric Coating
Preparation of Ferroelectric BaTiO3 Coating by Hydrothermal Synthesis
Basic Characterization of the Substrate and Coating
Cell Seeding
Evaluation of Cell Number and Cell Population Doubling Time
Evaluation of the Intensity of Fluorescence
Measurement of the Size of Cell Adhesion Area
2.10. Statistical Analysis
Basic Properties of The Batio3 Film and Ti39Nb Substrate
XRD Analysis of Ti39Nb Substrates and BaTiO3 Film
Raman Spectroscopy of Ti39Nb Substrates and BaTiO3 Film
Procedure Ba
Specific Markers of Adhesion and Differentiation in Cells on the Materials
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