Multi and single walled carbon nanotubes: effects on cell responses and biomineralization of osteoblasts cultures.

The use of carbon nanotubes (CNTs) on the development of biomaterials has been motivated by their excellent mechanical properties that could improve synthetic bone materials. However, the toxicity of CNTs on the tissue/implant interface and their influence on the biomineralization process have some contradictions. We investigated the influence of CNTs on osteoblasts plated on titanium (Ti) discs or plastic surfaces. We evaluated osteoblasts viability, alkaline phosphatase (ALP) activity, and mineralized matrix formation in the different phases of osteoblasts growth in the presence of single-walled CNTs (SWCNTs) and multi-walled CNTs (MWCNTs). An increase in osteoblasts viability was observed at the 21st day for both CNTs on plastic surface, while viability increased for MWCNTs at the 7th and 14th days and at the 7th day for SWCNTs on Ti discs compared to control. ALP activity increased at the 14th and 21st days for MWCNTs on plastic surfaces. For cells incubated with SWCNTs, an increase in ALP activity at the 7th day for plastic surface and at the 14th day for both materials (plastic and Ti) was observed. The mineralized matrix formation increased at the 21st day on plastic surface with SWCNTs, and at the 14th and 21st days for both CNTs on Ti discs. In conclusion, both SWCNTs and MWCNTs are not toxic to osteoblasts at concentrations up to 5 × 10(-5) and 1.3 × 10(-2) mg/mL, respectively, either in Ti discs or plastic surfaces. In the long term, the cells grown in contact with both CNTs and Ti presented better results regarding bone-like nodules formation.

Graphene oxide and titanium: synergistic effects on the biomineralization ability of osteoblast cultures.

Graphene oxide (GO) has attracted remarkable attention in recent years due to properties such as extremely large surface area, biocompatibility, biostability, and easy chemical functionalization. Osteoblasts underlie the deposition of hydroxyapatite crystals in the bone protein matrix during biomineralization; hydroxyapatite deposition involves extracellular matrix vesicles that are rich in alkaline phosphatase (ALP). Here, we have investigated how GO affects osteoblast viability, ALP activity, and mineralized matrix formation in osteoblast cultures in three different phases of cell growth, in the presence and in the absence of titanium (Ti). Scanning electron microscopy (SEM), Raman spectra, and energy dispersive spectroscopy aided GO characterization. The presence of GO increased the viability of osteoblast cells grown on a plastic surface. However, osteoblast viability on Ti discs was lower in the presence than in the absence of GO. ALP activity emerged at 14 days for the cell culture incubated with GO. The total protein concentration also increased at 21 days on both the Ti discs and plastic surface. Osteoblasts grown on Ti discs had increased mineralized matrix formation in the presence of GO as compared to the cells grown in the absence of GO. SEM images of the cell cultures on plastic surfaces in the presence of GO suggested delayed mineralized matrix formation. In conclusion, applications requiring the presence of Ti, such as prostheses and implants, should benefit from the use of GO, which may increase mineralized nodule formation, stimulate biomineralization, and accelerate bone regeneration.

Different compact hybrid Langmuir-Blodgett-film coatings modify biomineralization and the ability of osteoblasts to grow.

Calcium phosphates (CaPs) are biomaterials widely used in tissue regeneration with outstanding biological performance. Although the tremendous improvements achieved in CaP's materials research over the years, their interaction with physiological environments still need to be fully understood. The aim of this study is to explore a biomimetic Langmuir-Blodgett (LB) membrane to template the growth of hydroxyapatite (HAp) coatings on Ti surfaces and the ability of these coatings in inducing biomineralization by osteoblasts cultured in vitro. Changing the phospholipids (i.e., dihexadecyl phosphate (DHP) or octadecylphosphonic acid (OPA)), we also tuned the surface Ca2+ concentration. This structural feature gave rise to different LB-hybrid surfaces where the concentration of Ca2+ in the OPA/HAp was higher than the concentration of Ca2+ in DHP/HAp coating. The higher Ca2+ amount on OPA/HAp coatings, allied to the physical-chemical features, lead to different responses on osteoblasts, stimulating or inhibiting the natural biomineralization. The OPA/HAp coating caused a delay in the osteoblast proliferation as indicated by the decrease in the cell viability at the 7th culture day. Improved cell differentiation triggered by the DHP/HAp coating resulted in higher osteoblast biomineralization. The present data underscore that besides both coatings being composed by HAp, the final interfacial composition and physical-chemical properties influence differently the osteoblast behavior. Although the best osteoblast's viability was found to OPA/HAp, our dataset attested that DHP/HAp induced mineralization more effectively than that. This unexpected finding highlight the importance of deeply understanding the biomaterial interface and suggest a promising approach to the design of biofunctional LB-based coatings with tunable properties. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2524-2534, 2018.

Formation of carbonated hydroxyapatite films on metallic surfaces using dihexadecyl phosphate-LB film as template.

Hydroxyapatite serves as a bioactive material for biomedical purposes, because it shares similarities with the inorganic part of the bone. However, how this material deposits on metallic surfaces using biomimetic matrices remains unclear. In this study, we deposited dihexadecyl phosphate, a phospholipid that bears a simple chemical structure, on stainless steel and titanium surfaces using the Langmuir-Blodgett (LB) technique; we employed the resulting matrix to grow carbonated hydroxyapatite. We obtained the calcium phosphate coating via a two-step process: we immersed the surfaces modified with the LB films into phosphate buffer, and then, we exposed the metal to a solution that simulated the concentration of ions in the human plasma. The latter step generated carbonated hydroxyapatite, the same mineral existing in the bone. The free energy related to the surface roughness and composition increased after we modified the supports. We investigated the film morphology by scanning electron and atomic force microscopies and determined surface composition by infrared spectroscopy and energy dispersive X-ray. We also studied the role of the surface roughness and the surface chemistry on cell viability. The surface-modified Ti significantly increased osteoblastic cells proliferation, supporting the potential use of these surfaces as osteogenic materials.