Trojan-Like Internalization of Anatase Titanium Dioxide Nanoparticles by Human Osteoblast Cells.

Dentistry and orthopedics are undergoing a revolution in order to provide more reliable, comfortable and long-lasting implants to patients. Titanium (Ti) and titanium alloys have been used in dental implants and total hip arthroplasty due to their excellent biocompatibility. However, Ti-based implants in human body suffer surface degradation (corrosion and wear) resulting in the release of metallic ions and solid wear debris (mainly titanium dioxide) leading to peri-implant inflammatory reactions. Unfortunately, our current understanding of the biological interactions with titanium dioxide nanoparticles is still very limited. Taking this into consideration, this study focuses on the internalization of titanium dioxide nanoparticles on primary bone cells, exploring the events occurring at the nano-bio interface. For the first time, we report the selective binding of calcium (Ca), phosphorous (P) and proteins from cell culture medium to anatase nanoparticles that are extremely important for nanoparticle internalization and bone cells survival. In the intricate biological environment, anatase nanoparticles form bio-complexes (mixture of proteins and ions) which act as a kind of 'Trojan-horse' internalization by cells. Furthermore, anatase nanoparticles-induced modifications on cell behavior (viability and internalization) could be understand in detail. The results presented in this report can inspire new strategies for the use of titanium dioxide nanoparticles in several regeneration therapies.

Importance of dynamic culture for evaluating osteoblast activity on dense silicon-substituted hydroxyapatite.

This paper reports an investigation on human osteoblast-like cells (SaOs-2) seeded onto pure hydroxyapatite (HA) and silicon-substituted HA (SiHA) tablets under static and dynamic culture conditions. The biological characterizations were conducted in classical static conditions in multi-wells plates, and in a perfusion bioreactor that permits continuous circulation of culture medium at 2 mL/h. The morphology, proliferation and differentiation of osteoblastic cells were examined for the two types of samples in the both culture conditions after 1, 3 and 8 days. Under dynamic conditions, cells cultured on SiHA surfaces showed a faster adhesion process and the formation of longer and thinner focal adhesions than in static conditions. The number of cells grown onto both ceramic surfaces was higher in dynamic conditions when compared with static conditions. Moreover, a higher activity of alkaline phosphatase was found for cells seeded under dynamic conditions. Our findings suggest that the application of perfusion culture system on cells cultured on dense substrates is valuable for predicting in vivo behaviour of cells on biomaterials.

Surface transformation of silicon-doped hydroxyapatite immersed in culture medium under dynamic and static conditions.

A comparative study of in vitro bioactivity of hydroxyapatite (HA) and silicon-doped hydroxyapatite (SiHA) has been carried out by immersion in a cell culture medium with or without fetal bovine serum during 14 days in static and dynamic conditions. A specific bioreactor was developed for the experiments in dynamic conditions. Ceramic surface transformations were characterized by electron microscopy, atomic force microscopy and X-ray photoelectron spectroscopy before and after immersion. The monitoring of calcium, phosphate and proteins in immersion medium was also done during the experiment. The two hydroxyapatite surfaces immersed in cell culture medium under dynamic conditions were found to be more probably covered by a new Mg-enriched Ca-deficient apatite layer than surfaces immersed under static conditions. These results suggest that dynamic procedure and medium with serum macromolecules seem to be more adequate to predict the in vivo activity of bioceramics. Moreover, SiHA presented a higher capacity of protein adsorption.

Chemical and topographical influence of hydroxyapatite and beta-tricalcium phosphate surfaces on human osteoblastic cell behavior.

The objective of this work was to evaluate the relative role of the calcium phosphate surface chemistry and surface topography on human osteoblast behavior. Highly dense phosphate ceramics (single-phase hydroxyapatite HA and beta-tricalcium phosphates TCP) presenting two distinct nano roughnesses were produced. Some samples were gold-sputter coated in order to conveniently mask the surface chemical effects (without modification of the original roughness) and to study the isolated effect of surface topography on cellular behavior. Our results indicated that the nanotopography of the studied ceramics had no effect on the cellular adhesion (cell spreading, focal contacts and stress fibers formation). On the contrary, strong topographical effects were verified on cell proliferation and differentiation. Moreover, the phosphate chemistry was responsible for changes in adhesion, proliferation and cell differentiation. On TCP, it was shown that the main influent parameter was surface chemistry, which negatively affected the initial cell adhesion but positively affected the subsequent stage of proliferation and differentiation. On HA, the main influent parameter was surface topography, which increased cell differentiation but lowered proliferation.

Surface energy of hydroxyapatite and beta-tricalcium phosphate ceramics driving serum protein adsorption and osteoblast adhesion.

The main objective of this work was to evaluate the specific role of calcium phosphates surface energy on serum protein adsorption and human osteoblast adhesion, by isolating chemical effects from those caused by topography. Highly dense phosphate ceramics (single-phase hydroxyapatite HA and beta-tricalcium phosphates beta-TCP) presenting two distinct nano roughnesses were produced. Some samples were gold-sputter coated in order to conveniently mask the surface chemical effects (without modification of the original roughness) and to study the isolated effect of surface topography on cellular behavior. The results indicated that the nano topography of calcium phosphates strongly affected the protein adsorption process, being more important than surface chemistry. The seeding efficacy of osteoblasts was not affected nor by the topography neither by the calcium phosphate chemistries but the beta-TCP chemistry negatively influenced cell spreading. We observed that surface hydrophobicity is another way to change protein adsorption on surfaces. The decrease of the polar component of surface energy on gold-coated samples leaded to a decreased albumin and fibronectin adsorption but to an increased cell adhesion. Overall, this work contributes to better understand the role of topography and surface chemistry of calcium phosphates in serum protein adsorption and osteoblast adhesion.