Karla S. Brammer

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 (≈30-nm diameter) nanotubes promoted adhesion without noticeable differentiation, whereas larger (≈70- to 100-nm diameter) nanotubes elicited a dramatic stem cell elongation (≈10-fold increased), which induced cytoskeletal stress and selective differentiation into osteoblast-like cells, offering a promising nanotechnology-based route for uniqueorthopedics-related hMSC treatments.

Improved bone-forming functionality on diameter-controlled TiO2 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.

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.

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.

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.

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.

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.

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.

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.

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.