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1.  Nanoporous membranes for medical and biological applications 
Synthetic nanoporous materials have numerous potential biological and medical applications that involve sorting, sensing, isolating and releasing biological molecules. Nanoporous systems engineered to mimic natural filtration systems are actively being developed for use in smart implantable drug delivery systems, bioartificial organs, and other novel nano-enabled medical devices. Recent advances in nanoscience have made it possible to precisely control the morphology as well as physical and chemical properties of the pores in nanoporous materials that make them increasingly attractive for regulating and sensing transport at the molecular level. In this work, an overview of nanoporous membranes for biomedical applications is given. Various in vivo and in vitro membrane applications, including biosensing, biosorting, immunoisolation and drug delivery, are presented. Different types of nanoporous materials and their fabrication techniques are discussed with an emphasis on membranes with ordered pores. Desirable properties of membranes used in implantable devices, including biocompatibility and antibiofouling behavior, are discussed. The use of surface modification techniques to improve the function of nanoporous membranes is reviewed. Despite the extensive research carried out in fabrication, characterization, and modeling of nanoporous materials, there are still several challenges that must be overcome in order to create synthetic nanoporous systems that behave similarly to their biological counterparts.
PMCID: PMC3684197  PMID: 20049818
Biosensing; Drug delivery; Implantable materials; Nanopores; Nano-scale membranes
2.  Ordered nanoporous silica as carriers for improved delivery of water insoluble drugs: a comparative study between three dimensional and two dimensional macroporous silica 
The goal of the present study was to compare the drug release properties and stability of the nanoporous silica with different pore architectures as a matrix for improved delivery of poorly soluble drugs. For this purpose, three dimensional ordered macroporous (3DOM) silica with 3D continuous and interconnected macropores of different sizes (200 nm and 500 nm) and classic mesoporous silica (ie, Mobil Composition of Matter [MCM]-41 and Santa Barbara Amorphous [SBA]-15) with well-ordered two dimensional (2D) cylindrical mesopores were successfully fabricated and then loaded with the model drug indomethacin (IMC) via the solvent deposition method. Scanning electron microscopy (SEM), N2 adsorption, differential scanning calorimetry (DSC), and X-ray diffraction (XRD) were applied to systematically characterize all IMC-loaded nanoporous silica formulations, evidencing the successful inclusion of IMC into nanopores, the reduced crystallinity, and finally accelerated dissolution of IMC. It was worth mentioning that, in comparison to 2D mesoporous silica, 3DOM silica displayed a more rapid release profile, which may be ascribed to the 3D interconnected pore networks and the highly accessible surface areas. The results obtained from the stability test indicated that the amorphous state of IMC entrapped in the 2D mesoporous silica (SBA-15 and MCM-41) has a better physical stability than in that of 3DOM silica. Moreover, the dissolution rate and stability of IMC loaded in 3DOM silica was closely related to the pore size of macroporous silica. The colorimetric 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Cell Counting Kit (CCK)-8 assays in combination with direct morphology observations demonstrated the good biocompatibility of nanoporous silica, especially for 3DOM silica and SBA-15. The present work encourages further study of the drug release properties and stability of drug entrapped in different pore architecture of silica in order to realize their potential in oral drug delivery.
PMCID: PMC3808157  PMID: 24174875
3D ordered macroporous silica; mesoporous silica; poorly soluble drugs; in vitro dissolution; stability test; in vitro cytotoxicity
3.  Development of a nanoporous and multilayer drug-delivery platform for medical implants 
Biodegradable polymers can be applied to a variety of implants for controlled and local drug delivery. The aim of this study is to develop a biodegradable and nanoporous polymeric platform for a wide spectrum of drug-eluting implants with special focus on stent-coating applications. It was synthesized by poly(DL-lactide-co-glycolide) (PLGA 65:35, PLGA 75:25) and polycaprolactone (PCL) in a multilayer configuration by means of a spin-coating technique. The antiplatelet drug dipyridamole was loaded into the surface nanopores of the platform. Surface characterization was made by atomic force microscopy (AFM) and spectroscopic ellipsometry (SE). Platelet adhesion and drug-release kinetic studies were then carried out. The study revealed that the multilayer films are highly nanoporous, whereas the single layers of PLGA are atomically smooth and spherulites are formed in PCL. Their nanoporosity (pore diameter, depth, density, surface roughness) can be tailored by tuning the growth parameters (eg, spinning speed, polymer concentration), essential for drug-delivery performance. The origin of pore formation may be attributed to the phase separation of polymer blends via the spinodal decomposition mechanism. SE studies revealed the structural characteristics, film thickness, and optical properties even of the single layers in the triple-layer construct, providing substantial information for drug loading and complement AFM findings. Platelet adhesion studies showed that the dipyridamole-loaded coatings inhibit platelet aggregation that is a prerequisite for clotting. Finally, the films exhibited sustained release profiles of dipyridamole over 70 days. These results indicate that the current multilayer phase therapeutic approach constitutes an effective drug-delivery platform for drug-eluting implants and especially for cardiovascular stent applications.
PMCID: PMC3469098  PMID: 23071394
drug delivery; implants; stents; polymers; spin-coating; atomic force microscopy
4.  Calcified Nanostructured Silicon Wafer Surfaces for Biosensing: Effects of Surface Modification on Bioactivity 
Disease Markers  2003;18(4):159-165.
The growth of known biologically-relevant mineral phases on semiconducting surfaces is one strategy to explicitly induce bioactivity in such materials, either for sensing or drug delivery applications. In this work, we describe the use of a spark ablation process to fabricate deliberate patterns of Ca10(PO4)6(OH)2 on crystalline Si (calcified nanoporous silicon). These patterns have been principally characterized by scanning electron microscopy in conjunction with elemental characterization by energy dispersive x-ray analysis. This is followed by a detailed comparison of the effects of fibroblast adhesion and proliferation onto calcified nanoporous Si, calcified nanoporous Si derivatized with alendronate, as well as control samples of an identical surface area containing porous SiO2. Fibroblast adhesion and proliferation assays demonstrate that a higher density of cells grow on the Ca3(PO4)2 /porous Si/ SiO2 structures relative to the alendronate-modified surfaces and porous Si/SiOM2 samples.
PMCID: PMC3850822  PMID: 12590169
Calcium phosphate; silicon; fibroblasts; biosensor
5.  Electrically facilitated translocation of protein through solid nanopore 
Nanoscale Research Letters  2014;9(1):140.
Nanopores have been proven as versatile single-molecule sensors for individual unlabeled biopolymer detection and characterization. In the present work, a relative large nanopore with a diameter of about 60 nm has been used to detect protein translocation driven by a series of applied voltages. Compared with previous studied small nanopores, a distinct profile of protein translocation through a larger nanopore has been characterized. First, a higher threshold voltage is required to drive proteins into the large nanopore. With the increase of voltages, the capture frequency of protein into the nanopore has been markedly enhanced. And the distribution of current blockage events is characterized as a function of biased voltages. Due to the large dimension of the nanopore, the adsorption and desorption phenomenon of proteins observed with a prolonged dwell time has been weakened in our work. Nevertheless, the protein can still be stretched into an unfolded state by increased electric forces at high voltages. In consideration of the high throughput of the large nanopore, a couple of proteins passing through the nanopore simultaneously occur at high voltage. As a new feature, the feasibility and specificity of a nanopore with distinct geometry have been demonstrated for sensing protein translocation, which broadly expand the application of nanopore devices.
PMCID: PMC3976542  PMID: 24661490
Protein translocation; Solid state nanopore; Current blockage; Translocation time
6.  Directly Observing the Motion of DNA Molecules near Solid-State Nanopores 
ACS nano  2012;6(11):10090-10097.
We investigate the diffusion and the drift motion of λ DNA molecules near solid-state nanopores prior to their translocation though the nanopores using fluorescence microscopy. The radial dependence of the electric field near a nanopore generated by an applied voltage in ionic solution can be estimated quantitatively in 3D by analyzing the motion of negatively charged DNA molecules. We find that the electric field is approximately spherically symmetric around the nanopore under the conditions investigated. In addition, DNA clogging at the nanopore was directly observed. Surprisingly, the probability of the clogging event increases with increasing external bias voltage. We also find that DNA molecules clogging the nanopore reduce the electric field amplitude at the nanopore membrane surface. To better understand these experimental results, analytical method with Ohm’s law and computer simulation with Poisson and Nernst-Planck (PNP) equations are used to calculate the electric field near the nanopore. These results are of great interest in both experimental and theoretical considerations of the motion of DNA molecules near voltage-biased nanopores. These findings will also contribute to the development of solid-state nanopore based DNA sensing devices.
PMCID: PMC3508321  PMID: 23046052
single-molecule; sensing; nanopore; DNA
7.  Direct Prototyping of Patterned Nanoporous Carbon: A Route from Materials to On-chip Devices 
Scientific Reports  2013;3:2294.
Prototyping of nanoporous carbon membranes with three-dimensional microscale patterns is significant for integration of such multifunctional materials into various miniaturized systems. Incorporating nano material synthesis into microelectronics technology, we present a novel approach to direct prototyping of carbon membranes with highly nanoporous structures inside. Membranes with significant thicknesses (1 ~ 40 μm) are rapidly prototyped at wafer level by combining nano templating method with readily available microfabrication techniques, which include photolithography, high-temperature annealing and etching. In particular, the high-surface-area membranes are specified as three-dimensional electrodes for micro supercapacitors and show high performance compared to reported ones. Improvements in scalability, compatibility and cost make the general strategy promising for batch fabrication of operational on-chip devices or full integration of three-dimensional nanoporous membranes with existing micro systems.
PMCID: PMC3724177  PMID: 23887486
8.  Ocular Biocompatibility and Structural Integrity of Micro- and Nanostructured Poly(caprolactone) Films 
The identification of biomaterials that are well tolerated in the eye is important for the development of new ocular drug delivery devices and implants, and the application of micro- and nanoengineered devices to biomedical treatments is predicated on the long-term preservation within the target organ or tissue of the very small functional design elements. This study assesses the ocular tolerance and durability of micro- and nanostructured biopolymer thin films injected or implanted into the rabbit eye. Structured poly(caprolactone) (PCL) thin films were placed in adult rabbit eyes for survival studies, with serial ophthalmic examinations over 6 months. Morphologic abnormalities and device/tissue reactions were evaluated by histologic studies, and scanning electron microscopy (SEM) of films was used to determine the structural integrity. Structured PCL thin films (20- to 40-μm thick) were constructed to design specifications with 50-μm linear microgrooves or arrays of nanopores with ∼30-nm diameters. After up to 9 months of ocular residency, SEM on devices retrieved from the eye showed preservation of micro- and nanostructural features. In ocular safety evaluations carried out over 6 months, serial examinations in 18 implanted eyes showed no evidence of chronic inflammation, cataractogenesis, or retinal toxicity. Postoperative ocular inflammation was seen in 67% of eyes for 1 week, and persistent corneal edema occurred in 1 eye. Histology revealed no ocular inflammation or morphologic abnormalities of ocular tissues. Thin-film/tissue responses such as cellular reaction, fibrosis, or surface biodeposits were not seen. Micro- and nanostructured PCL thin films exhibited acceptable ocular tolerance and maintained the structural integrity of design features while residing in the eye. Thin-film micro- and nanostructured PCL appears to be a feasible biomaterial for intraocular therapeutic applications.
PMCID: PMC3601720  PMID: 23391326
9.  Regulating the Transport of DNA through Biofriendly Nanochannels in a Thin Solid Membrane 
Scientific Reports  2014;4:3985.
Channels formed by membrane proteins regulate the transport of water, ions or nutrients that are essential to cells' metabolism. Recent advances in nanotechnology allow us to fabricate solid-state nanopores for transporting and analyzing biomolecules. However, uncontrollable surface properties of a fabricated nanopore cause irregular transport of biomolecules, limiting potential biomimetic applications. Here we show that a nanopore functionalized with a self-assembled monolayer (SAM) can potentially regulate the transport of a DNA molecule by changing functional groups of the SAM. We found that an enhanced interaction between DNA and a SAM-coated nanopore can slow down the translocation speed of DNA molecules and increase the DNA capture-rate. Our results demonstrate that the transport of DNA molecules inside nanopores could be modulated by coating a SAM on the pore surface. Our method to control the DNA motion inside a nanopore may find its applications in nanopore-based DNA sequencing devices.
PMCID: PMC3914175  PMID: 24496378
10.  A Single-Molecule Nanopore Device Detects DNA Polymerase Activity With Single-Nucleotide Resolution 
The ability to monitor DNA polymerase activity with single-nucleotide resolution has been the cornerstone of a number of advanced single-molecule DNA sequencing concepts. Toward this goal, we report the first spatially-resolved observation of DNA polymerase activity with single-base resolution at the single-molecule level. We describe the design and characterization of a single-species supramolecular nanopore device capable of detecting up to nine consecutive DNA polymerase-catalyzed single nucleotide primer extensions with high sensitivity and spatial resolution (≤ 2.4 Å). The device is assembled in a suspended lipid membrane by threading and mechanically capturing a single strand of DNA-PEG copolymer inside an α-hemolysin protein pore. Single nucleotide primer extensions result in successive displacements of the template DNA strand within the protein pore, which can be monitored by the corresponding stepped changes in the ion current flowing through the pore under an applied transmembrane potential. The system described thus represents a promising advance toward nanopore-mediated single-molecule DNA sequencing concept, and in addition might be applicable to studying a number of other biopolymer-protein interactions and dynamics.
PMCID: PMC2453067  PMID: 18166054
11.  Synthesis and Characterization of Polydiacetylene Films and Nanotubes 
We report here the synthesis and characterization of polydiacetylene (PDA) films and nanotubes using layer-by-layer (LBL) chemistry. 10,12-Docosadiyndioic acid (DCDA) monomer was self-assembled on flat surfaces and inside of nanoporous alumina templates. UV irradiation of DCDA provided polymerized-DCDA (PDCDA) films and nanotubes. We have used zirconium-carboxylate interlayer chemistry to synthesize PDCDA multilayers on flat surfaces and in nanoporous template. PDCDA multilayers were characterized using optical (UV–vis, fluorescence, ellipsometry, FTIR) spectroscopies, ionic current–voltage (I–V) analysis, and scanning electron microscopy. Ellipsometry, FTIR, electronic absorption and emission spectroscopies showed a uniform DCDA deposition at each deposition cycle. Our optical spectroscopic analysis indicates that carboxylate-zirconium interlinking chemistry is robust. To explain the disorganization in the alkyl portion of PDCDA multilayer films, we propose carboxylate-zirconium interlinkages act as “locks” in between PDCDA layers which restrict the movement of alkyl portion in the films. Because of this locking, the induced-stresses in the polymer chains can not be efficiently relieved. Our ionic resistance data from I–V analysis correlate well with calculated resistance at smaller number of PDCDA layers but significantly deviated for thicker PDCDA nanotubes. These differences were attributed to ion-blocking because some of the PDCDA nanotubes were totally closed and the nonohmic and permselective ionic behaviors when the diameter of the pores approaches the double-layer thickness of the solution inside of the nanotubes.
PMCID: PMC2683165  PMID: 18823090
12.  Assessing Graphene Nanopores for Sequencing DNA 
Nano letters  2012;12(8):4117-4123.
Using all-atom molecular dynamics and atomic-resolution Brownian dynamics, we simulate the translocation of single-stranded DNA through graphene nanopores and characterize the ionic current blockades produced by DNA nucleotides. We find that transport of single DNA strands through graphene nanopores may occur in single nucleotide steps. For certain pore geometries, hydrophobic interactions with the graphene membrane lead to a dramatic reduction in the conformational fluctuations of the nucleotides in the nanopores. Furthermore, we show that ionic current blockades produced by different DNA nucleotides are, in general, indicative of the nucleotide type, but very sensitive to the orientation of the nucleotides in the nanopore. Taken together, our simulations suggest that strand sequencing of DNA by measuring the ionic current blockades in graphene nanopores may be possible, given that the conformation of DNA nucleotides in the nanopore can be controlled through precise engineering of the nanopore surface.
PMCID: PMC3434709  PMID: 22780094
Nanopore; graphene; molecular dynamics; biosensors; nucleic acids; ionic current
13.  Automated Forward and Reverse Ratcheting of DNA in a Nanopore at Five Angstrom Precision1 
Nature biotechnology  2012;30(4):344-348.
Single-molecule techniques have been developed for commercial DNA sequencing1,2. One emerging strategy uses a nanopore to analyze DNA molecules as they are driven electrophoretically in single file order past a sensor3-5. However, uncontrolled DNA strand electrophoresis through nanopores is too fast for accurate base reads6. A proposed solution would employ processive enzymes to deliver DNA through the pore at a slower average rate7. Here, we describe forward and reverse ratcheting of DNA templates through the α–hemolysin (α-HL) nanopore controlled by wild-type phi29 DNA polymerase (phi29 DNAP). DNA strands were examined in single file order at one nucleotide spatial precision in real time. The registry error probability (either an insertion or deletion during one pass along a template strand) ranged from 10% to 24.5% absent optimization. This general strategy facilitates multiple reads of individual template strands and is transferrable to other nanopore devices for implementation of DNA sequence analysis.
PMCID: PMC3408072  PMID: 22334048
14.  Nanopore-based Sensors for Multi-parameter Characterization of Nanoparticles and Viruses 
Nanopore-based instrumentation has been developed for improved multi-parameter characterization of the physical properties of nano- and micro-sized particles. Accurate measurement, with individual particle resolution, of a range of biological and synthetic sample types, i.e. liposomes, PLGA, lipids, micelles, virus-like-particles, polymers, viruses, bacteria, protein-conjugates, exosomes, and vesicles will be presented.
Particles are transported through a size-tunable pore via electric field and/or with pressure, for rapid and detailed determination of particle concentration (particles/mL for dosage), accurate size, aggregation levels, size distribution and relative surface charge distribution, all determined simultaneously.
Experimental parameters are adjusted in real-time for mapping how different populations within particle mixtures respond to externally applied conditions for high-resolution and powerful analysis of particle physical properties and their dynamic behaviour, i.e. to assess the level of surface modification (PEG-lyation) of drug delivery carriers.
The ability to individually interrogate each particle addresses the shortcomings of ensemble systems such as dynamic light-scattering and also of static systems using electron microscopy. This also enables the quantification of the dynamic behaviour of particle mixtures, such as aggregation and fragmentation of particles, and surface modification changes to particles.
Research work utilizing tunable pore sensors in virus quantification, pathogen interaction dynamics, medical diagnostics and drug delivery systems are presented.
PMCID: PMC3630705
15.  Infuence of Microstructure in Drug Release Behavior of Silica Nanocapsules 
Journal of Drug Delivery  2013;2013:803585.
Meso- and nanoporous structures are adequate matrices for controlled drug delivery systems, due to their large surface areas and to their bioactive and biocompatibility properties. Mesoporous materials of type SBA-15, synthesized under different pH conditions, and zeolite beta were studied in order to compare the different intrinsic morphological characteristics as pore size, pore connectivity, and pore geometry on the drug loading and release process. These materials were characterized by X-ray diffraction, nitrogen adsorption, scanning and transmission electron microscopy, and calorimetric measurements. Ibuprofen (IBU) was chosen as a model drug for the formulation of controlled-release dosage forms; it was impregnated into these two types of materials by a soaking procedure during different periods. Drug loading and release studies were followed by UV-Vis spectrophotometry. All nano- and mesostructured materials showed a similar loading behavior. It was found that the pore size and Al content strongly influenced the release process. These results suggest that the framework structure and architecture affect the drug adsorption and release properties of these materials. Both materials offer a good potential for a controlled delivery system of ibuprofen.
PMCID: PMC3748776  PMID: 23986870
16.  Conductance-Based Determination of Solid-State Nanopore Size and Shape: An Exploration of Performance Limits 
Knowledge of nanopore size and shape is critical for many implementations of these single-molecule sensing elements. Geometry determination by fitting the electrolyte-concentration-dependence of the conductance of surface-charged, solid-state nanopores has been proposed to replace demanding electron microscope-based methods. The functional form of the conductance poses challenges for this method by restricting the number of free parameters used to characterize the nanopore. We calculated the electrolyte-dependent conductance of nanopores with an exponential-cylindrical radial profile using three free geometric parameters; this profile, itself, could not be uniquely geometry-optimized by the conductance. Several different structurally simplified models, however, generated quantitative agreement with the conductance, but with errors exceeding 40% for estimates of key geometrical parameters. A tractable conical-cylindrical model afforded a good characterization of the nanopore size and shape, with errors of less than 1% for the limiting radius. Understanding these performance limits provides a basis for using and extending analytical nanopore conductance models.
PMCID: PMC3673737  PMID: 23750286
Electric double layer; nanopore surface charge; nanopore conductance; nanopore shape; silicon nitride nanopore; silicon oxide nanopore
17.  Enhanced Microcontact Printing of Proteins on Nanoporous Silica Surface 
Nanotechnology  2010;21(41):415302.
We demonstrate porous silica surface modification, combined with microcontact printing, as an effective method for enhanced protein patterning and adsorption on arbitrary surfaces. Compared to conventional chemical treatments, this approach offers scalability and long-term device stability without requiring complex chemical activation. Two chemical surface treatments using functionalization with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA) were compared with the nanoporous silica surface on the basis of protein adsorption. The deposited thickness and uniformity of the porous silica films were evaluated for fluorescein isothiocyanate (FITC)-labeled rabbit immunoglobulin G (R-IgG) protein printed onto the substrates via patterned polydimethlysiloxane (PDMS) stamps. A more complete transfer of proteins was observed on porous silica substrates compared to chemically functionalized substrates. A comparison of different pore sizes (2–6 nm), and porous silica thicknesses (30–200 nm) indicates that porous silica with 4 nm diameter, 57% porosity and a thickness of 96 nm provided a suitable environment for complete transfer of R-IgG proteins. Both fluorescence microscopy and atomic force microscopy (AFM) were used for protein layers characterizations. A porous silica layer is biocompatible, providing a favorable transfer medium with minimal damage to the proteins. A patterned immunoassay microchip was developed to demonstrate the retained protein function after printing on nanoporous surfaces, which enables printable and robust immunoassay detection for point-of-care applications.
PMCID: PMC2944042  PMID: 20834118
Porous sílica; microcontact printing; Polydimethylsiloxane (PDMS); Proteins; IgG
18.  Nanopore-Induced Spontaneous Concentration for Optofluidic Sensing and Particle Assembly 
Analytical chemistry  2012;85(2):971-977.
Metallic nanopore arrays have emerged as optofluidic platforms with multifarious sensing and analytical capabilities such as label-free surface plasmon resonance (SPR) sensing of molecular binding interactions and surface-enhanced Raman spectroscopy (SERS). However, directed delivery of analytes through open nanopores using traditional methods such as external electric fields or pressure gradients still remains difficult. We demonstrate that nanopore arrays have an intrinsic ability to promote flow through them via capillary flow and evaporation. This passive “nano-drain” mechanism is utilized to concentrate biomolecules on the surface of nanopores for improved detection sensitivity or create ordered nanoscale arrays of beads and liposomes. Without using any external pump or fluidic interconnects, we can concentrate and detect the presence of less than a femtomole of streptavidin in 10 µL of sample using fluorescence imaging. Liposome nanoarrays are also prepared in less than 5 minutes and used to detect lipid-protein interactions. We also demonstrate label-free SPR detection of analytes using metallic nanopore arrays. This method provides a fast, simple, transportable and small-volume platform for labeled as well as label-free plasmonic analysis while improving the detection time and sensitivity.
PMCID: PMC3568508  PMID: 23214989
Nanohole array; nanopore; microfluidics; nanofluidics; surface plasmon resonance; flow-through sensing; diffusion limits; optofluidics; plasmonics; liposome; fluidic self-assembly
19.  Fibronectin and vitronectin promote human fetal osteoblast cell attachment and proliferation on nanoporous titanium surfaces 
Improvements in osteoconduction of implant biomaterials require focusing on the bone-implant interface, which is a complex multifactorial system. Surface topography of implants plays a crucial role at this interface. Nanostructured surfaces have been shown to promote serum protein adsorption and osteoblast adhesion when compared to microstructured surfaces for bone-implant materials. We studied the influence of the serum proteins fibronectin and vitronectin on the attachment and proliferation of osteoblasts onto nanostructured titania surfaces. Human fetal osteoblastic cells hFOB 1.19 were used as model osteoblasts and were grown on nanoporous TiO2 templates, using Ti6Al4V and commercially pure Ti substrates as controls. Results show a significant increase in cell proliferation on nanoporous TiO2 over flat substrates. Initial cell attachment data exhibited a significant effect by either fibronectin or vitronectin on cell adhesion at the surface of any of the tested materials. In addition, the extent of cell adhesion was significantly different between the nanoporous TiO2 and both Ti6Al4V and commercially pure Ti substrates, with the first showing the highest surface coverage. There was no significant difference on osteoblast attachment or proliferation between the presence of fibronectin or vitronectin using any of the material substrates. Taken together, these results suggest that the increase in osteoblast attachment and proliferation shown on the nanoporous TiO2 is due to an increase in the adsorption of fibronectin and vitronectin because of the higher surface area and to an enhanced protein unfolding, which allows access to osteoblast binding motifs within these proteins.
PMCID: PMC3718483  PMID: 23858975
nanoporous TiO2; hFOB 1.19; protein adsorption; vitronectin; fibronectin
20.  DNA characterization with Ion Beam Sculpted Silicon Nitride Nanopores 
Solid state nanopores are emerging as robust single molecule electronic measurement devices and as platforms for confining biomolecules for further analysis. The first silicon nitride nanopore to detect individual DNA molecules were fabricated using ion beam sculpting (IBS), a method that uses broad, low energy ion beams to create nanopores with dimensions ranging from 2 to 20 nm. In this chapter, we discuss the fabrication, characterization, and use of IBS sculpted nanopores as well as efficient uses of pClamp and MATLAB software suites for data acquisition and analysis. The fabrication section will cover the repeatability and the pore size limits. The characterization discussion focuses on the geometric properties as measured by low and high resolution transmission electron microscopy (TEM), electron energy loss spectroscopy (EELS), and energy filtered TEM (EFTEM). The section on translocation experiments focuses on how to use tools commonly available to the nanopore experimenter to determine whether a pore will be useful for experimentation or if it should be abandoned. A memory efficient method of taking data using Clampex’s event-driven mode and dual channel recording will be presented, followed by an easy to implement multi-threshold event detection and classification method using MATLAB software.
PMCID: PMC3727399  PMID: 22528259
Ion beam sculpting; silicon nitride nanopore; ionic current blockage; DNA size; DNA conformation
21.  The Targeted Delivery of Multicomponent Cargos to Cancer Cells via Nanoporous Particle-Supported Lipid Bilayers 
Nature Materials  2011;10(5):389-397.
Encapsulation of drugs within nanocarriers that selectively target malignant cells promises to mitigate side effects of conventional chemotherapy and to enable delivery of the unique drug combinations needed for personalized medicine. To realize this potential, however, targeted nanocarriers must simultaneously overcome multiple challenges, including specificity, stability, and a high capacity for disparate cargos. Here we report porous nanoparticle-supported lipid bilayers (protocells) that synergistically combine properties of liposomes and nanoporous particles. Protocells modified with a targeting peptide that binds to human hepatocellular carcinoma (HCC) exhibit a 10,000-fold greater affinity for HCC than for hepatocytes, endothelial cells, and immune cells. Furthermore, protocells can be loaded with combinations of therapeutic (drugs, siRNA, and toxins) and diagnostic (quantum dots) agents and modified to promote endosomal escape and nuclear accumulation of selected cargos. The enormous capacity of the high-surface-area nanoporous core combined with the enhanced targeting efficacy enabled by the fluid supported lipid bilayer allow a single protocell loaded with a drug cocktail to kill a drug-resistant HCC cell, representing a 106-fold improvement over comparable liposomes.
PMCID: PMC3287066  PMID: 21499315
22.  Pulsed Laser Deposition of Nanoporous Cobalt Thin Films 
Nanoporous cobalt thin films were deposited on anodized aluminum oxide (AAO) membranes at room temperature using pulsed laser deposition. Scanning electron microscopy demonstrated that the nanoporous cobalt thin films retained the monodisperse pore size and high porosity of the anodized aluminum oxide substrates. Temperature- and field-dependent magnetic data obtained between 10 K and 350 K showed large hysteresis behavior in these materials. The increase of coercivity values was larger for nanoporous cobalt thin films than for multilayered cobalt/alumina thin films. The average diameter of the cobalt nanograins in the nanoporous cobalt thin films was estimated to be ~5 nm for blocking temperatures near room temperature. These results suggest that pulsed laser deposition may be used to fabricate nanoporous magnetic materials with unusual properties for biosensing, drug delivery, data storage, and other technological applications.
PMCID: PMC3690133  PMID: 19198344
A. Magnetization curves; B. Epitaxial films; C. Magnetic nano-networks
23.  Antibacterial hemostatic dressings with nanoporous bioglass containing silver 
Nanoporous bioglass containing silver (n-BGS) was fabricated using the sol-gel method, with cetyltrimethyl ammonium bromide as template. The results showed that n-BGS with nanoporous structure had a surface area of 467 m2/g and a pore size of around 6 nm, and exhibited a significantly higher water absorption rate compared with BGS without nanopores. The n-BGS containing small amounts of silver (Ag) had a slight effect on its surface area. The n-BGS containing 0.02 wt% Ag, without cytotoxicity, had a good antibacterial effect on Escherichia coli, and its antibacterial rate reached 99% in 12 hours. The n-BGS’s clotting ability significantly decreased prothrombin time (PT) and activated partial thromboplastin time (APTT), indicating n-BGS with a higher surface area could significantly promote blood clotting (by decreasing clotting time) compared with BGS without nanopores. Effective hemostasis was achieved in skin injury models, and bleeding time was reduced. It is suggested that n-BGS could be a good dressing, with antibacterial and hemostatic properties, which might shorten wound bleeding time and control hemorrhage.
PMCID: PMC3383339  PMID: 22745538
antibacterial; bioglass; cytotoxicity; dressing; hemostasis; nanopore; silver
24.  Fabrication and centeracterization of ordered CuIn(1−x)GaxSe2 nanopore films via template-based electrodeposition 
Nanoscale Research Letters  2012;7(1):675.
Ordered CuIn(1−x)GaxSe2 (CIGS) nanopore films were prepared by one-step electrodeposition based on porous anodized aluminum oxide templates. The as-grown film shows a highly ordered morphology that reproduces the surface pattern of the substrate. Raman spectroscopy and X-ray diffraction pattern show that CIGS nanopore films had ideal chalcopyrite crystallization. Energy dispersive spectroscopy reveals the Cu-Se phases firstly formed in initial stage of growth. Then, indium and gallium were incorporated in the nanopore films in succession. Cu-Se phase is most likely to act as a growth promoter in the growth progress of CIGS nanopore films. Due to the high surface area and porous structure, this kind of CIGS films could have potential application in light-trapping CIGS solar cells and photoelectrochemical water splitting.
PMCID: PMC3552846  PMID: 23245846
CuIn(1−x)GaxSe2; nanopore films; electrodeposition; anodic aluminumoxide; annealing; 82.45.Yz; 81.05.Rm; 81.15.Pq; 81.40.Ef
25.  A Macroporous TiO2 Oxygen Sensor Fabricated Using Anodic Aluminium Oxide as an Etching Mask 
Sensors (Basel, Switzerland)  2010;10(1):670-683.
An innovative fabrication method to produce a macroporous Si surface by employing an anodic aluminium oxide (AAO) nanopore array layer as an etching template is presented. Combining AAO with a reactive ion etching (RIE) processes, a homogeneous and macroporous silicon surface can be effectively configured by modulating AAO process parameters and alumina film thickness, thus hopefully replacing conventional photolithography and electrochemical etch methods. The hybrid process integration is considered fully CMOS compatible thanks to the low-temperature AAO and CMOS processes. The gas-sensing characteristics of 50 nm TiO2 nanofilms deposited on the macroporous surface are compared with those of conventional plain (or non-porous) nanofilms to verify reduced response noise and improved sensitivity as a result of their macroporosity. Our experimental results reveal that macroporous geometry of the TiO2 chemoresistive gas sensor demonstrates 2-fold higher (∼33%) improved sensitivity than a non-porous sensor at different levels of oxygen exposure. In addition, the macroporous device exhibits excellent discrimination capability and significantly lessened response noise at 500 °C. Experimental results indicate that the hybrid process of such miniature and macroporous devices are compatible as well as applicable to integrated next generation bio-chemical sensors.
PMCID: PMC3270862  PMID: 22315561
anodic aluminium oxide (AAO); macroporous; MEMS; TiO2 gas sensor

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