Lingzhou Zhao

Antibacterial Coatings on Titanium Implants

Titanium and titanium alloys are key biomedical materials because of their good biocompatibility and mechanical properties. Nevertheless, infection on and around titanium implants still remains a problem which is usually difficult to treat and may lead to eventual implant removal. As a result, preventive measures are necessary to mitigate implant-frelated infection. One important strategy is to render the implant surface antibacterial by impeding the formation of a biofilm. A number of approaches have been proposed for this purpose and they are reviewed in this article.

Antibacterial nano-structured titania coating incorporated with silver nanoparticles.

Titanium (Ti) implants are widely used clinically but post-operation infection remains one of the most common and serious complications. A surface boasting long-term antibacterial ability is highly desirable in order to prevent implant associated infection. In this study, titania nanotubes (TiO(2)-NTs) incorporated with silver (Ag) nanoparticles are fabricated on Ti implants to achieve this purpose. The Ag nanoparticles adhere tightly to the wall of the TiO(2)-NTs prepared by immersion in a silver nitrate solution followed by ultraviolet light radiation. The amount of Ag introduced to the NTs can be varied by changing processing parameters such as the AgNO(3) concentration and immersion time. The TiO(2)-NTs loaded with Ag nanoparticles (NT-Ag) can kill all the planktonic bacteria in the suspension during the first several days, and the ability of the NT-Ag to prevent bacterial adhesion is maintained without obvious decline for 30 days, which are normally long enough to prevent post-operation infection in the early and intermediate stages and perhaps even late infection around the implant. Although the NT-Ag structure shows some cytotoxicity, it can be reduced by controlling the Ag release rate. The NT-Ag materials are also expected to possess satisfactory osteoconductivity in addition to the good biological performance expected of TiO(2)-NTs. This controllable NT-Ag structure which provides relatively long-term antibacterial ability and good tissue integration has promising applications in orthopedics, dentistry, and other biomedical devices.

The influence of hierarchical hybrid micro/nano-textured titanium surface with titania nanotubes on osteoblast functions

Hierarchical hybrid micro/nano-textured titanium surface topographies with titania nanotubes were produced by simple acid etching followed by anodization to mimic the hierarchical structure of bone tissues. Primary rat osteoblasts were used to evaluate the bioactivity. The microtopography formed by acid etching of titanium induced inconsistent osteoblast functions with initial cell adhesion and osteogenesis-related gene expression being dramatically enhanced while other cell behaviors such as proliferation, intracellular total protein synthesis and alkaline phosphatase activity, collagen secretion, and extracellular matrix mineralization being depressed. In comparison, addition of nanotubes to the microtopography led to enhancement of multiple osteoblast functions. Nearly all the cell functions investigated in this study were retained or promoted. Compared to a microtopography, the enhancement of multiple cell functions observed from the hierarchical micro/nano-textured surfaces is expected to lead to faster bone maturation around the titanium implants without compromising the bone mass. In addition, the hierarchical micro/nano-textured surfaces still retain the mechanical interlocking ability of the microtopography thereby boding well for osseointegration. Our study reveals a synergistic role played by the micro and nanotopographies in osteoblast functions and provides insight to the design of better biomedical implant surfaces.

Effects of micropitted/nanotubular titania topographies on bone mesenchymal stem cell osteogenic differentiation

Micro/nanotopographical modification of biomaterials constitutes a promising approach to direct stem cell osteogenic differentiation to promote osseointegration. In this work, titania nanotubes (NTs) 25 and 80 nm in size with the acid-etched Ti topography (AcidTi) and hierarchical hybrid micropitted/nanotubular topographies (Micro/5VNT and Micro/20VNT) are produced to mimic the structure of the natural bone extracellular matrix (ECM). The effects on bone mesenchymal stem cell (MSC) osteogenic differentiation are studied systematically by various microscopic and biological characterization techniques. Cell adhesion is assayed by nucleus fluorescence staining and cell proliferation is studied by CCK-8 assay and flow cytometry. Osteogenic differentiation is assayed by alkaline phosphatase (ALP) expression, collagen secretion, matrix mineralization, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis on the osteogenesis related gene expression. All the topographies are observed to induce MSC osteogenic differentiation in the absence of osteogenic supplements. The nanotube surfaces significantly promote cell attachment and spread, collagen secretion and ECM mineralization, as well as osteogenesis-related gene expression. Among them, Micro/20VNT shows the best ability to simultaneously promote MSC proliferation and osteogenic differentiation. Our results unambiguously demonstrate their excellent ability to support MSC proliferation and induce MSC osteogenic differentiation, especially those with the micropitted topography.

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 effects and biocompatibility of titanium surfaces with graded silver incorporation in titania nanotubes.

Most commercial dental implants are made of titanium (Ti) because Ti possesses excellent properties such as osseointegration. However, many types of Ti products still suffer from insufficient antibacterial capability and bacterial infection after surgery remains one of the most common and intractable complications. In this study, a dual process encompassing anodization and silver plasma immersion ion implantation (Ag PIII) is utilized to produce titania nanotubes (TiO₂-NTs) containing Ag at different sites and depths. The concentration and depth of the incorporated Ag can be tailored readily by changing the PIII parameters. The Ag-embedded TiO₂-NTs which retain the nanotubular morphology are capable of sterilizing oral pathogens as opposed to pure Ti plates and pristine TiO₂-NTs. Biological assays indicate that the in vitro and in vivo biocompatibility of the sample plasma-implanted at a lower voltage of 0.5 kV (NT-Ag-0.5) is significantly compromised due to the large amount of surface Ag. On the other hand, the sample implanted at 1 kV (NT-Ag-1.0) exhibits unimpaired effects due to the smaller surface Ag accumulation. Sample NT-Ag-1.0 is further demonstrated to possess sustained antibacterial properties due to the large embedded depth of Ag and the technique and resulting materials have large potential in dental implants.

Osteogenic activity and antibacterial effects on titanium surfaces modified with Zn-incorporated nanotube arrays

Titanium implants having enhanced osteogenic activity and antibacterial property are highly desirable for the prevention of implant associated infection and promotion of osseointegration. In this study, coatings containing titania nanotubes (NTs) incorporated with zinc (NT-Zn) are produced on Ti implants by anodization and hydrothermal treatment in Zn containing solutions. The amount of incorporated Zn can be adjusted by varying the structural parameters such as the nanotube diameter and length as well as hydrothermal treatment time. The suitable NT-Zn coatings with good intrinsic antibacterial properties can prevent post-operation infection. Excellent osteogenesis inducing ability in the absence of extraneous osteogenic supplements is demonstrated and the ERK1/2 signaling is found to be involved. The NT-Zn structure which is simple, stable, and easy to produce and scale up has immense potential in bone implant applications.

The role of sterilization in the cytocompatibility of titania nanotubes.

Titiania nanotubes have large potential in medical implant applications but their tissue compatibility is still controversial. Since the sterilization methods may impact the biocompatibility of titania nanotubes and be the source of the controversy, we investigate the influence of three commonly used sterilization methods, autoclaving, ultraviolet irradiation and ethanol immersion, on the cytocompatibility of titania nanotubes. Two titania nanostructures, namely nanonets with an average pore diameter of 25 nm and nanotubes with an average diameter of 80 nm, are used in this study. The results show that the sterilization methods significantly affect the cytocompatibility of these titania surfaces. UV and ethanol sterilization give rise to a higher surface free energy inducing higher initial cell adhesion and proliferation compared to autoclaving, whereas UV irradiation produces the best cell functions including adhesion, proliferation, as well as differentiation represented by related gene expressions. The cytocompatibility results obtained from the nanoscale surfaces are compared to those acquired from the polished surface demonstrating the significant effects. Our results suggest that the sterilization process plays an important role in the observed cytocompatibility of titania nanotubes and may be the reason for the controversial results so far. UV sterilization is found to be the best method from the viewpoint of surface contamination elimination.

Improved implant osseointegration of a nanostructured titanium surface via mediation of macrophage polarization.

The use of endosseous implanted materials is often limited by undesirable effects that may be due to macrophage-related inflammation. The purpose of this study was to fabricate a nanostructured surface on a titanium implant to regulate the macrophage inflammatory response and improve the performance of the implant. Anodization at 5 and 20 V as well as UV irradiation were used to generate hydrophilic, nanostructured TiO2 surfaces (denoted as NT5 and NT20, respectively). Their surface characteristics and in vivo osseointegration as well as the inflammatory response they elicit were analyzed. In addition, the behavior of macrophages in vitro was evaluated. Although the in vitro osteogenic activity on the two surfaces was similar, the NT5 surface was associated with more bone formation, less inflammation, and a reduced CD68(+) macrophage distribution in vivo compared to the NT20 and polished Ti surfaces. Consistently, further experiments revealed that the NT5 surface induced healing-associated M2 polarization in vitro and in vivo. By contrast, the NT20 surface promoted the pro-inflammatory M1 polarization, which could further impair bone regeneration. The results demonstrate the dominant role of macrophage-related inflammation in bone healing around implants and that surface nanotopography can be designed to have an immune-regulating effect in support of the success of implants.

Fabrication, modification, and biomedical applications of anodized TiO2 nanotube arrays

Titanium dioxide (TiO2) nanotubes have attracted increasing attention due to their outstanding properties and potential applications in photocatalysis, dye-sensitized solar cells, and biomedical devices. In this paper, recent research progress on TiO2 nanotube arrays (NTAs) produced by anodic oxidation of Ti in fluoride-containing electrolytes is reviewed with emphasis on the modification methods and biomedical applications. The fabrication protocol and growth mechanism are first discussed and common modification methods used to improve the optical, electronic, and biomedical properties of TiO2 NTAs are reviewed. Photo/electro-chemical biosensors based on TiO2 NTAs dedicated to the detection of glucose, hydrogen peroxide, and other biomolecules are described and recent examples of using TiO2 NTAs to improve the cellular response in vitro and accelerate osseointegration in vivo are provided. The incorporation and delivery of inorganic bioactive agents such as Ag, Sr, and Zn to achieve antibacterial and/or osteogenesis inducing ability are described and finally, the outlook and future development of TiO2 nanotubes pertaining to biomedical devices are briefly discussed.