Ruiqiang Hang

The effects of titania nanotubes with embedded silver oxide nanoparticles on bacteria and osteoblasts.

A versatile strategy to endow biomaterials with long-term antibacterial ability without compromising the cytocompatibility is highly desirable to combat biomaterial related infection. TiO2 nanotube (NT) arrays can significantly enhance the functions of many cell types including osteoblasts thus having promising applications in orthopedics, orthodontics, as well as other biomedical fields. In this study, TiO2 NT arrays with Ag2O nanoparticle embedded in the nanotube wall (NT-Ag2O arrays) are prepared on titanium (Ti) by TiAg magnetron sputtering and anodization. Well-defined NT arrays containing Ag concentrations in a wide range from 0 to 15 at % are formed. Ag incorporation has little influence on the NT diameter, but significantly decreases the tube length. Crystallized Ag2O nanoparticles with diameters ranging from 5 nm to 20 nm are embedded in the amorphous TiO2 nanotube wall and this unique structure leads to controlled release of Ag(+) that generates adequate antibacterial activity without showing cytotoxicity. The NT-Ag2O arrays can effectively kill Escherichia coli and Staphylococcus aureus even after immersion for 28 days, demonstrating the long lasting antibacterial ability. Furthermore, the NT-Ag2O arrays have no appreciable influence on the osteoblast viability, proliferation, and differentiation compared to the Ag free TiO2 NT arrays. Ag incorporation even shows some favorable effects on promoting cell spreading. The technique reported here is a versatile approach to develop biomedical coatings with different functions.

Antibacterial activity and cytocompatibility of Cu–Ti–O nanotubes

TiO2 nanotubes (NTs) have favorable biological properties, but the poor antibacterial activity limits their application especially in orthopedics fields. In this article, Cu-Ti-O nanotubes with different Cu contents are fabricated on sputtered TiCu films. Scanning electron microscopy reveals the NTs can be formed on sputtered TiCu films when the Cu content is less than 14.6 at %. X-ray photoelectron spectroscopy results indicate the NTs are consist of CuO mixed with TiO2 and the Cu content in NTs decreases dramatically compared with that in TiCu films. Biological experiments show that although these NTs have poor release antibacterial activity, their contact antibacterial activity has proven to be excellent, indicating the NT surface can effectively inhibit biomaterial-associated infections. The cytocompatibility of the NTs is closely related to the Cu content and when its content is relatively low (1.01 at %), there is no appreciable cytotoxicity. So Cu-Ti-O NTs with 1 at % Cu may be suitable to achieve proper antibacterial activity and desired cytocompatibility. The Cu-Ti-O NTs integrate the favorable antibacterial activity of Cu and excellent biological properties of TiO2 NTs therefore have potential applications in orthopedics, dentistry, and other biomedical fields.

A nano-silver composite based on the ion-exchange response for the intelligent antibacterial applications

As a kind of antimicrobial agent, nano-silver composites have attracted a great deal of interest in biomedical applications. However, the typical loadings of silver nanoparticles (AgNPs) in such composites could result in dose-related cytotoxicity. In this study, a nano-silver composite leading to antimicrobial activity without cytotoxicity was fabricated by loading AgNPs into a dried alginate hydrogel. The biological performance of this composite mainly depended on the release of AgNPs, which needed to be triggered by the ion-exchange response and was further influenced by the loadings of AgNPs in the composite. The antimicrobial activity against E. coli and S. aureus demonstrated that the released silver no less than 678 ppb in the medium caused a reduction of 7log10CFU/mL (100%) bacteria. Significantly, the dose (~1.10×10(3) ppb) of released silver was not toxic and allowed attachment, and growth of MC3T3-E1 pre-osteoblast cells. These results supported that the composite was compatible with in vitro mammalian cells yet exhibited antimicrobial activity by carefully designing the loadings of AgNPs within the alginate. Thus, it indicated that the performance of this composite might permit management of bacterial infection in wound beds without impairment of wound healing.

Microstructure and cytotoxicity evaluation of duplex-treated silver-containing antibacterial TiO2 coatings

Implant-related infection is one of the most common and serious complications associated with biomedical implantation. To prevent bacterial adhesion, a series of porous TiO2 coatings with different concentrations of silver (designated as M0, M1, M2 and M3) were prepared on pure titanium substrates by a duplex-treatment technique combining magnetron sputtering with micro-arc oxidation. All coatings are porous with pore size less than 5 μm and the concentrations of silver in the M0, M1, M2 and M3 are 0, 0.95, 1.36 and 1.93 wt.%, respectively. Silver is found to be distributed throughout the thickness of the coatings by scanning electron microscopy. The release of silver from the TiO2 coatings was confirmed by an inductively-coupled plasma mass spectroscopy. The antibacterial effects of these coatings were tested against Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli), and the cytotoxicity was evaluated using the mouse pre-osteoblast cells. The results indicate that the antibacterial activities of TiO2 coatings are greatly improved due to the incorporation of silver. No cytotoxic effect is found for the M1 surfaces from the observation of pre-osteoblast cell by MTT assay and fluorescence microscopy. Although the M2 and M3 coatings appeared to be toxic for pre-osteoblast cells after 1 day in culture, the cell viability on M2 and M3 surfaces was greatly raised after culturing for 2 days. Our results suggested that the TiO2 coatings incorporated with an optimum amount of silver can possess excellent antibacterial activities without cytotoxic effect, which has promising applications in biomedical devices.

Highly ordered Ni-Ti-O nanotubes for non-enzymatic glucose detection.

Anodization is used to fabricate Ni-Ti-O nanotube (NT) electrodes for non-enzymatic glucose detection. The morphology, microstructure and composition of the materials are characterized by field emission scanning electron microscopy (FE-SEM), high resolution transmission electron microscopy (HR-TEM) and X-ray photoelectron spectroscopy (XPS). Our results show amorphous and highly ordered NTs with diameter of 50nm, length of 800nm, and Ni/Ti ratio (at %) of 0.35 can be fabricated in ethylene glycol electrolyte supplemented with 0.2 wt.% NH4F and 0.5 vol.% H2O at 30°C and 25V for 1h. Electrochemical experiments indicate that at an applied potential of 0.60V vs. Ag/AgCl, the electrode exhibits a linear response window for glucose concentrations from 0.002mM to 0.2mM with a response time of 10s, detection limit of 0.13μM (S/N=3), and sensitivity of 83μAmM(-1)cm(-2). The excellent performance of the electrode is attributed to its large specific area and fast electron transfer between the NT walls. The good electrochemical performance of the Ni-Ti-O NTs as well as their simple and low-cost preparation method make the strategy promising in non-enzymatic glucose detection.

Fabrication of Ni-Ti-O nanotube arrays by anodization of NiTi alloy and their potential applications.

Nickel-titanium-oxide (Ni-Ti-O) nanotube arrays (NTAs) prepared on nearly equiatomic NiTi alloy shall have broad application potential such as for energy storage and biomedicine, but their precise structure control is a great challenge because of the high content of alloying element of Ni, a non-valve metal that cannot form a compact electronic insulating passive layer when anodized. In the present work, we systemically investigated the influence of various anodization parameters on the formation and structure of Ni-Ti-O NTAs and their potential applications. Our results show that well controlled NTAs can be fabricated during relatively wide ranges of the anodization voltage (5–90 V), electrolyte temperature (10–50°C) and electrolyte NH4F content (0.025–0.8 wt%) but within a narrow window of the electrolyte H2O content (0.0–1.0 vol%). Through modulating these parameters, the Ni-Ti-O NTAs with different diameter (15–70 nm) and length (45–1320 nm) can be produced in a controlled manner. Regarding potential applications, the Ni-Ti-O NTAs may be used as electrodes for electrochemical energy storage and non-enzymic glucose detection, and may constitute nanoscaled biofunctional coating to improve the biological performance of NiTi based biomedical implants.

Osteogenic and angiogenic activities of silicon-incorporated TiO2 nanotube arrays

Osteogenesis and angiogenesis that have interaction in vivo are two pivotal processes for implant osseointegration, so implant surfaces with both enhanced osteogenic and angiogenic activities are in need. Developing silicon (Si) doped TiO2 nanotube array (TNA-Si) coatings shall be a promising strategy to yield favorable implant osseointegration with the combined effects of TiO2 nanotube arrays (TNAs) and Si in the enhancement of both osteogenic and angiogenic activities. To achieve this purpose, TNA-Sis are fabricated through the unique strategy of anodization of magnetron-sputtered TiSi coatings. Under optimized conditions, a highly ordered nanotubular structure with even dispersion of Si throughout the nanotubes in the form of SiO2 can be obtained. Si incorporation has little influence on the nanotube length, but slightly decreases the diameter, thickens the nanotube wall, and increases the hydrophilicity. TNA-Sis show good cytocompatibility to both osteoblasts and endothelial cells (ECs). TNA-Sis show enhanced proliferation, spreading, alkaline phosphatase activity, collagen secretion, and matrix mineralization of osteoblasts. Meanwhile, TNA-Sis induce better EC proliferation, and the conditioned culture media from TNA-Sis generate better angiogenic ability, nitric oxide production, and vascular endothelial growth factor secretion from ECs. In particular, TNA-Si4.6 with the strongest osteogenic and angiogenic activities in the context of the present study is highly promising as the next-generation hard tissue implant coating.

Antibacterial ability and angiogenic activity of Cu-Ti-O nanotube arrays

Bacterial infection and loosening of orthopedic implants remain two disastrously postoperative complications. Angiogenesis is critical important to facilitate implant osseointegration in vivo. TiO2 nanotubes arrays (NTAs) with proper dimensions possess good osseointegration ability. Accordingly, the present work incorporated copper (Cu) into TiO2 NTAs (Cu-Ti-O NTAs) to enhance their antibacterial ability and angiogenesis activity, which was realized through anodizing magnetron-sputtered TiCu coatings with different Cu contents on pure titanium (Ti). Our results show ordered Cu-Ti-O NTAs can be produced under proper Cu content (<15.14%) in TiCu coatings. The NTAs possess excellent long-term antibacterial ability against Staphylococcus aureus (S. aureus), which may be ascribed to sustained release of Cu2+. The cytotoxicity of Cu-Ti-O NTAs to endothelial cells (ECs) could be negligible and can even promote cell proliferation as revealed by live/dead staining and MTT. Meanwhile, Cu-Ti-O NTAs can up-regulate nitric oxide (NO) synthesis and vascular endothelial growth factors (VEGF) secretion of ECs on the sample surfaces compared with that of pure TiO2 NTAs (control). Furthermore, the angiogenic activity is also enhanced in ionic extracts of Cu-Ti-O NTAs compared with the control. The excellent long-term antibacterial ability and favorable angiogenic activity render Cu-Ti-O NTAs to be promising implant coatings.

Antibacterial ability and osteogenic activity of porous Sr/Ag-containing TiO2 coatings.

Implant-associated infection and poor osseointegration remains a major clinical challenge in Ti-based implant materials. A versatile strategy to endow Ti-based implants with long-term antibacterial ability as well as better osteogenic activity is highly desirable for high quality implantation. Strontium (Sr) has been shown to be a significant element to favor bone growth by promoting new bone formation and inhibiting bone resorption. In this study, a novel duplex-treatment technique encompassing magnetron sputtering with micro-arc oxidation is utilized to fabricate porous Sr/Ag-containing TiO2 coatings loaded with different concentrations of Ag and Sr. All coatings are porous with pore size less than 5 µm. Ag is primarily distributed homogeneously inside the pores, and the concentrations of Ag in Sr/Ag-containing TiO2 coatings with low and high Ag contents are 0.40 at.% and 0.83 at.% respectively. We have demonstrated that this kind of coating displays long-lasting antibacterial ability even up to 28 d due to the incorporation of Ag. Further, Sr/Ag-containing TiO2 coatings with optimum Ag and Sr contents revealed good cytocompatibility, enhanced osteoblast spreading and osseointegration, which stemmed primarily from the synergistic effect exerted by the porous surface topography and the bioactive element Sr. However, this study has also identified, for the first time, that proper addition of Ag would further facilitate osteogenic effects. Besides, Sr may be able to alleviate the potential cytotoxic effect of excessive Ag. Thus, integration of optimum functional elements Ag and Sr into Ti-based implant materials would be expected to expedite osseointegration while simultaneously sustaining long-term antibacterial activity, which would provide new insights for relevant fundamental investigations and biomedical applications.

Biological response of endothelial cells to diamond‐like carbon‐coated NiTi alloy

Diamond-like carbon (DLC) coatings were deposited on nearly equiatomic nickel-titanium (NiTi) alloy by arc-enhanced magnetron sputtering. The microstructure, surface morphology, chemical composition, surface free energy, protein adsorbance, and leach amount of Ni ions were assessed by Raman spectroscopy, high-resolution transmission electron microscopy (HR-TEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), contact angle measurements, micro BCA™ protein assay kit, and inductively coupled plasma mass spectrometry (ICP-MS). The biological response of the endothelial cells (ECs) was evaluated by cell adhesion, morphology, viability, and expression levels of thrombogenicity-related genes. Our results show that the DLC coatings inhibit the release of Ni ions from the NiTi substrate effectively thus enhancing its biosafety. The easy adhesion, elongated morphology, and high viability of ECs on the DLC coatings suggest fast endothelialization after implantation and so application of DLC coatings improves the surface properties of NiTi in cardiovascular applications. The relationship between the surface characteristics, Ni leaching, and concomitant biological response are discussed in details.