Tantalum coating on TiO2 nanotubes induces superior rate of matrix mineralization and osteofunctionality in human osteoblasts.

Nanostructured surface geometries have been the focus of a multitude of recent biomaterial research, and exciting findings have been published. However, only a few publications have directly compared nanostructures of various surface chemistries. The work herein directly compares the response of human osteoblast cells to surfaces of identical nanotube geometries with two well-known orthopedic biomaterials: titanium oxide (TiO2) and tantalum (Ta). The results reveal that the Ta surface chemistry on the nanotube architecture enhances alkaline phosphatase activity, and promotes a ~30% faster rate of matrix mineralization and bone-nodule formation when compared to results on bare TiO2 nanotubes. This study implies that unique combinations of surface chemistry and nanostructure may influence cell behavior due to distinctive physico-chemical properties. These findings are of paramount importance to the orthopedics field for understanding cell behavior in response to subtle alterations in nanostructure and surface chemistry, and will enable further insight into the complex manipulation of biomaterial surfaces. With increased focus in the field of orthopedic materials research on nanostructured surfaces, this study emphasizes the need for careful and systematic review of variations in surface chemistry in concurrence with nanotopographical changes.

Controlled metallic nanopillars for low impedance biomedical electrode.

Radial metallic nanopillar/nanowire structures can be created by a controlled radiofrequency (RF) plasma processing technique on the surface of certain alloy wires, including important biomedical alloys such as MP35N (Co-Ni-Cr-Mo alloy), platinum-iridium and stainless steel. In electrode applications such as pacemakers or neural stimulators, the increase in surface area in elongated MP35N nanopillars allows for decreased surface impedance and greater current density. However, the nanopillar height on MP35N alloy tends to be self-limiting at ∼1-3µm. The objective of this study was to further elongate the radial nanopillars so as to reduce electrode impedance for biomedical electrode applications. Intelligent experimental design allowed for efficient investigation of processing parameters, including plasma material, process duration, power, pressure and repetition. It was found that multi-step repeated processing in the parameter-controlled RF environment could increase nanopillar height to ∼10µm, a 400% improvement, while the RF plasma processing with identical total duration but in a single step did not lead to desired nanopillar elongation. Measurement of electrode impedance in phosphate-buffered saline solution showed an associated decrease to one-fifth of the surface impedance of unprocessed wire for signals below 100Hz. For the purposes of this study, MP35N and Pt-Ir wires were characterized and demonstrated augmented surface impedance properties which, in combination with superior cell integration, enhanced biomedical electrode performance.

Variations to the nanotube surface for bone regeneration.

The complex mechanisms of the bone cell-surface interactions are yet to be completely understood, and researchers continue to strive to uncover the fully optimized implant material for perfect osseointegration. A particularly fascinating area of research involves the study of nanostructured surfaces, which are believed to enhance osteogenic behavior, possibly due to the mimicry of components of the extracellular matrix of bone. There is a growing body of data that emphasizes the promise of the titanium oxide (TiO2) nanotube architecture as an advanced orthopedic implant material. The review herein highlights findings regarding TiO2 nanotube surfaces for bone regeneration and the osteogenic effects of minute changes to the surface such as tube size and surface chemistry.

Hybrid micro/nano-topography of a TiO2 nanotube-coated commercial zirconia femoral knee implant promotes bone cell adhesion in vitro.

Various approaches have been studied to engineer the implant surface to enhance bone in-growth properties, particularly using micro- and nano-topography. In this study, the behavior of osteoblast (bone) cells was analyzed in response to a titanium oxide (TiO2) nanotube-coated commercial zirconia femoral knee implant consisting of a combined surface structure of a micro-roughened surface with the nanotube coating. The osteoblast cells demonstrated high degrees of adhesion and integration into the surface of the nanotube-coated implant material, indicating preferential cell behavior on this surface when compared to the bare implant. The results of this brief study provide sufficient evidence to encourage future studies. The development of such hierarchical micro- and nano-topographical features, as demonstrated in this work, can provide insightful designs for advanced bone-inducing material coatings on ceramic orthopedic implant surfaces.

TiO2 nanotubes for bone regeneration.

Nanostructured materials are believed to play a fundamental role in orthopedic research because bone itself has a structural hierarchy at the first level in the nanometer regime. Here, we report on titanium oxide (TiO(2)) surface nanostructures utilized for orthopedic implant considerations. Specifically, the effects of TiO(2) nanotube surfaces for bone regeneration will be discussed. This unique 3D tube shaped nanostructure created by electrochemical anodization has profound effects on osteogenic cells and is stimulating new avenues for orthopedic material surface designs. There is a growing body of data elucidating the benefits of using TiO(2) nanotubes for enhanced orthopedic implant surfaces. The current trends discussed within foreshadow the great potential of TiO(2) nanotubes for clinical use.

Comparative cell behavior on carbon-coated TiO2 nanotube surfaces for osteoblasts vs. osteo-progenitor cells.

Surface engineering approaches that alter the topological chemistry of a substrate could be used as an effective tool for directing cell interactions and their subsequent function. It is well known that the physical environment of nanotopography has positive effects on cell behavior, yet direct comparisons of nanotopographic surface chemistry have not been fully explored. Here we compare TiO(2) nanotubes with carbon-coated TiO(2) nanotubes, probing osteogenic cell behavior, including osteoblast (bone cells) and mesenchymal stem cell (MSC) (osteo-progenitor cells) interactions with the different surface chemistries (TiO(2) vs. carbon). The roles played by the material surface chemistry of the nanotubes did not have an effect on the adhesion, growth or morphology, but had a major influence on the alkaline phosphatase (ALP) activity of osteoblast cells, with the original TiO(2) chemistry having higher ALP levels. In addition, the different chemistries caused different levels of osteogenic differentiation in MSCs; however, it was the carbon-coated TiO(2) nanotubes that had the greater advantage, with higher levels of osteo-differentiation. It was observed in this study that: (a) chemistry plays a role in cell functionality, such as ALP activity and osteogenic protein gene expression (PCR); (b) different cell types may have different chemical preferences for optimal function. The ability to optimize cell behavior using surface chemistry factors has a profound effect on both orthopedic and tissue engineering in general. This study aims to highlight the importance of the chemistry of the carrier material in osteogenic tissue engineering schemes.

Hydrophobic nanopillars initiate mesenchymal stem cell aggregation and osteo-differentiation.

Surface engineering approaches that alter the physical topography of a substrate could be used as an effective tool and as an alternative to biochemical means of directing stem cell interactions and their subsequent differentiation. In this paper we compare hydrophobic micro- vs. nanopillar type fabrication techniques for probing mesenchymal stem cell (MSC) interaction with the surface physical environment. The roles played by the topography of the nanopillar in particular influenced MSC growth and allowed for regulatory control of the stem cell fate. The nanopillar induced large 3-D cell aggregates to form on the surface which had up-regulated osteogenic specific matrix components. The ability to control MSC differentiation, using only the topographical factors, has a profound effect on both MSC biology and tissue engineering. This study aims to highlight the importance of the physical material carrier in stem cell based tissue engineering schemes.

Magnetically vectored nanocapsules for tumor penetration and remotely switchable on-demand drug release.

Nanocapsules containing intentionally trapped magnetic nanoparticles and defined anticancer drugs have been prepared to provide a powerful magnetic vector under moderate gradient magnetic fields. These nanocapsules can penetrate into the interior of tumors and allow a controlled on-off switchable release of the drug cargo via remote RF field. This smart drug delivery system is compact as all the components can be self-contained in 80-150 nm capsules. In vitro as well as in vivo results indicate that these nanocapsules can be enriched near the mouse breast tumor and are effective in reducing tumor cell growth.

Plasma-induced nanopillars on bare metal coronary stent surface for enhanced endothelialization.

An increased risk of late stent thrombosis associated with polymer carriers on the surface of drug-eluting stents remains one of the challenges in cardiovascular stent technology, which has instigated a renewed interest in the polymer-less, bare metal stent approach. As thrombus formation is most likely augmented by the lack of endothelial cell coverage at the exposed stented site, an improved stent surface that enhances cell coverage is essential for viable polymer-less all metal stents. We demonstrate superior endothelial cell growth, more continuous monolayer formation and overall improved endothelialization with nanopillar arrays created via radio frequency plasma surface texturing on our all metallic stent surface of MP35N stent alloy. It is shown that the nanotextured surface significantly up-regulates primary bovine aortic endothelial cell (BAEC) functionality when compared with unprocessed, smooth MP35N surfaces without a nanopillar topography. The desirable presence of transmembrane tight junctions and highly organized monolayer formation was induced by the presence of the nanopillar surface texture. The nanopillar structure also produced a reduced level of oxidative stress in the BAECs. These findings may contribute to new nanotechnology-based surface design concepts for bare metal stents producing advanced cardiovascular implants which mitigate late stent thrombosis.

Radially arrayed nanopillar formation on metallic stent wire surface via radio-frequency plasma.

MP35N (Co-Ni-Cr-Mo alloy) is an important stent implant material for which many aspects, that include nanostructured surfaces, are yet to be understood. The present study provides the first creation of radially emanating metallic nanopillar structures on the surface of MP35N stent alloy wires; a novel textured surface structuring derived via controlled RF processing technique. The goal of this study was to characterize the newly found structures, identify evolution stages of nanopillar formations, as well as optimize RF process parameters for controlled surface texturing technique for stent wire materials. The exposure of a stent alloy wire, 250 microm diameter Co-Ni-Cr-Mo alloy (MP35N), to parameter-controlled RF environment resulted in dense surface nanostructures consisting of high-aspect-ratio dendritic nanopillars/nanowires. Extensive surface characterization and local compositional analyses by Transmission Electron Microscopy (TEM), Energy Dispersive X-ray analysis (EDX) and X-ray photoelectron spectroscopy (XPS) show increased values of Mo contents on the outer edges of protruding nanopillars, indicating a possibility of the higher Mo content phase contributing to the differential plasma sputter etching on the MP35N surface and resultant nanowire formation. A comparative investigation on single phase alloy versus multi-phase alloy seems to point to the importance of phase segregation for successful nanowire formation by RF plasma treatment. In addition to MP35N, some specific single phased materials, such as Fe-Ni and Fe-Cr alloys or Pt metal wire, were exposed in same RF plasma conditions and results did not form the complex structures found on MP35N samples. For the purpose of this study, metallic stent wires that have nanostructured surfaces can be considered a "polymer-less" approach to surface modification, The creation and characterization of radially arrayed nanostructured surfaces has been demonstrated on MP35N stent alloy wires using this RF plasma process; where such nanostructured surfaces contribute to design concepts that may enhance endotheliazation of stent materials via surface texturing modification.

Titanium dioxide nanotubes enhance bone bonding in vivo.

Implant topography is critical to the clinical success of bone-anchored implants, yet little is known how nano-modified implant topography affects osseointegration. We investigate the in vivo bone bonding of two titanium implant surfaces: titanium dioxide (TiO(2)) nanotubes and TiO(2) gritblasted surfaces. In previous in vitro studies, the topography of the TiO(2) nanotubes improved osteoblast proliferation and adhesion compared with gritblasted titanium surfaces. After four weeks of implantation in rabbit tibias, pull-out testing indicated that TiO(2) nanotubes significantly improved bone bonding strength by as much as nine-fold compared with TiO(2) gritblasted surfaces. Histological analysis confirmed greater bone-implant contact area, new bone formation, and calcium and phosphorus levels on the nanotube surfaces. It is anticipated that further studies will contribute to a better understanding of the effect of implant nanotopography on in vivo bone formation and bonding strength.

Antibiofouling, sustained antibiotic release by Si nanowire templates.

Loading or filling nanostructures with antibiotics can be one of the relevant approaches for obtaining a controlled drug release rate. Vertically aligned silicon nanowire (SiNW) arrays with 10-40 nm diameter wires having 1-3 microm in length obtained by the electroless etching (EE) technique are used in this study as novel nanostructures for mediating drug delivery. Here we report controlled antibiotic activity and sustained bioavailability from SiNW arrays and also show microstructural manipulations for a tunable release rate. As well, we have demonstrated biodegradability of SiNWs in phosphate buffer saline (PBS) solution. Strikingly suppressed cell and protein adhesion was observed on our SiNW surface, which indicates a reduced probability for biofouling and drug release impediments. Such antibiotic release from the nanowire-structured surface can provide more reliable antibiotic protection at a targeted implantation or biosensor site.

Geometry transformation and alterations of periodically patterned Si nanotemplates by dry oxidation.

We report on the size-dependent transformation and geometrical modifications of periodically patterned Si templates by a combination of dry oxidation and chemical etching. Deep ultraviolet lithography patterned circular holes with diameters varying between 190 nm and 1 microm on Si wafers were oxidized at 1000 degrees C using dry oxygen for various durations, with selected samples chemically etched for oxide removal for additional alterations. An interesting phenomenon of a circular-to-square shape transformation of the holes was observed, which was particularly pronounced in the sub-200 nm regime. We tentatively attribute the change to the surface energy and geometry constraints in nanoscale patterns.

Improved bone-forming functionality on diameter-controlled TiO(2) nanotube surface.

The titanium dioxide (TiO(2)) nanotube surface enables significantly accelerated osteoblast adhesion and exhibits strong bonding with bone. We prepared various sizes (30-100 nm diameter) of titanium dioxide (TiO(2)) nanotubes on titanium substrates by anodization and investigated the osteoblast cellular behavior in response to these different nanotube sizes. The unique and striking result of this study is that a change in osteoblast behavior is obtained in a relatively narrow range of nanotube dimensions, with small diameter ( approximately 30 nm) nanotubes promoting the highest degree of osteoblast adhesion, while larger diameter (70-100 nm) nanotubes elicit a lower population of cells with extremely elongated cellular morphology and much higher alkaline phosphatase levels. Increased elongation of nuclei was also observed with larger diameter nanotubes. By controlling the nanotopography, large diameter nanotubes, in the approximately 100 nm regime, induced extremely elongated cellular shapes, with an aspect ratio of 11:1, which resulted in substantially enhanced up-regulation of alkaline phosphatase activity, suggesting greater bone-forming ability than nanotubes with smaller diameters. Such nanotube structures, already being a strongly osseointegrating implant material, offer encouraging implications for the development and optimization of novel orthopedics-related treatments with precise control toward desired cell and bone growth behavior.

Stem cell fate dictated solely by altered nanotube dimension.

Two important goals in stem cell research are to control the cell proliferation without differentiation and to direct the differentiation into a specific cell lineage when desired. Here, we demonstrate such paths by controlling only the nanotopography of culture substrates. Altering the dimensions of nanotubular-shaped titanium oxide surface structures independently allowed either augmented human mesenchymal stem cell (hMSC) adhesion or a specific differentiation of hMSCs into osteoblasts by using only the geometric cues, absent of osteogenic inducing media. hMSC behavior in response to defined nanotube sizes revealed a very dramatic change in hMSC behavior in a relatively narrow range of nanotube dimensions. Small (approximately 30-nm diameter) nanotubes promoted adhesion without noticeable differentiation, whereas larger (approximately 70- to 100-nm diameter) nanotubes elicited a dramatic stem cell elongation (approximately 10-fold increased), which induced cytoskeletal stress and selective differentiation into osteoblast-like cells, offering a promising nanotechnology-based route for unique orthopedics-related hMSC treatments.

Enhanced cellular mobility guided by TiO2 nanotube surfaces.

The in vitro endothelial response of primary bovine aortic endothelial cells (BAECs) was investigated on a flat Ti surface vs a nanostructured TiO2 nanotube surface. The nanotopography provided nanoscale cues that facilitated cellular probing, cell sensing, and especially cell migration, where more organized actin cytoskeletal filaments formed lamellipodia and locomotive morphologies. Motile cell protrusions were able to probe down into the nanotube pores for contact stimulation, and focal adhesions were formed and disassembled readily for enhanced advancement of cellular fronts, which was not observed on a flat substrate of titanium. NOx and endothelin-1 functional assays confirmed that the nanotubes also up-regulated an antithrombic cellular state for maintaining vascular tone. The enhanced endothelial response to TiO2 nanotubes is significant for a potential modification of vascular stent surfaces in order to increase the rate and reliability of endothelialization and endothelial cell migration onto the stent for repairing arterial injury after activation.