Assessing the Acidic and Alkaline Recalcitrance of Covalently Modified Surface Amines on Ordered Mesoporous Carbon.

Ordered mesoporous carbon (OMC) scaffolds were covalently modified with primary amine groups by means of oxidation-coupling, yielding C-O-C bonds, or organometallic activation-coupling, yielding C-C bonds. The aminated OMCs were stressed by immersion in either 1 M hydrochloric acid or 1 M sodium hydroxide solutions at room temperature for 6 h and characterized by nitrogen sorption, electron microscopy, low-angle X-ray diffraction, thermogravimetric analysis, and the 4-nitrobenzaldehyde assay. Results demonstrate that aminated surfaces of OMC by butyllithium grafting are stable toward both 1 M HCl and 1 M NaOH, whereas the oxidation-aminated OMC surfaces can withstand 1 M NaOH only. This study illustrates the importance of chemical testing to supplant chemical intuition when tailoring carbon surfaces for applications where strong acids or bases are employed. This is especially emphasized for carbonaceous materials because of the surface heterogeneity among different carbon allotropes.

Best Practices in the Characterization of MOF@MSN Composites.

Research on permanently porous nanomaterials has gripped the attention of materials chemists for decades. Mesoporous silica nanoparticles (MSNs) and metal-organic frameworks (MOFs) are two of the most studied classes of materials in this field. Recently, explorations into embedding MOFs within the mesopores of MSNs have aimed to create composites that are greater than the sum of their parts. While initial progress has been promising, it has become clear that the characterization of these MOF@MSN composite materials represents a significant challenge that is often overlooked, leading to an unfortunate ambiguity in the field. The greatest difficulty lies in determining whether the product of a synthesis is simply a physical mixture of the two materials or truly the targeted composite, with MOF exclusively crystallized in the pores or on the surfaces of the MSN. This challenge is aggravated by the dramatically different porosity and composition of the components, often resulting in ambiguous information from common characterization techniques. This Viewpoint will address this challenge by calling attention to the mentioned issues and proposing a standardized approach to characterizing these materials. In particular, the use of powder X-ray diffraction, gas physisorption, and electron microscopy with systematic control experiments and data analysis is outlined. This approach can provide the information needed to clearly validate the architecture of an apparent MOF@MSN composite.

Monitoring the Stimulated Uncapping Process of Gold-Capped Mesoporous Silica Nanoparticles.

To establish a new method for tracking the interaction of nanoparticles with chemical cleaving agents, we exploited the optical effects caused by attaching 5-10 nm gold nanoparticles with molecular linkers to large mesoporous silica nanoparticles (MSN). At low levels of gold loading onto MSN, the optical spectra resemble colloidal suspensions of gold. As the gold is removed, by cleaving agents, the MSN revert to the optical spectra typical of bare silica. Time-lapse images of gold-capped MSN stationed in microchannels reveal that the rate of gold release is dependent on the concentration of the cleaving agent. The uncapping process was also monitored successfully for MSN endocytosed by A549 cancer cells, which produce the cleaving agent glutathione. These experiments demonstrate that the optical properties of MSN can be used to directly monitor cleaving kinetics, even in complex cellular settings.

Conserved Activity of Reassociated Homotetrameric Protein Subunits Released from Mesoporous Silica Nanoparticles.

Mesoporous silica nanoparticles (MSN) with enlarged pores were prepared and characterized, and reversibly dissociated subunits of concanavalin A were entrapped in the mesopores, as shown by multiple biochemical and material characterizations. When loaded in the MSN, we demonstrated protein stability from proteases and, upon release, the subunits reassociated into active proteins shown through mannose binding and o-phthalaldehyde fluorescence. We have demonstrated a versatile and facile method to load homomeric proteins into MSN with potential applications in enhancing the delivery of large therapeutic proteins.

Organometallic Complexes Anchored to Conductive Carbon for Electrocatalytic Oxidation of Methane at Low Temperature.

Low-temperature direct methane fuel cells (DMEFCs) offer the opportunity to substantially improve the efficiency of energy production from natural gas. This study focuses on the development of well-defined platinum organometallic complexes covalently anchored to ordered mesoporous carbon (OMC) for electrochemical oxidation of methane in a proton exchange membrane fuel cell at 80 °C. A maximum normalized power of 403 µW/mg Pt was obtained, which was 5 times higher than the power obtained from a modern commercial catalyst and 2 orders of magnitude greater than that from a Pt black catalyst. The observed differences in catalytic activities for oxidation of methane are linked to the chemistry of the tethered catalysts, determined by X-ray photoelectron spectroscopy. The chemistry/activity relationships demonstrate a tangible path for the design of electrocatalytic systems for C-H bond activation that afford superior performance in DMEFC for potential commercial applications.

Mesoporous silica nanoparticle-mediated intracellular cre protein delivery for maize genome editing via loxP site excision.

The delivery of proteins instead of DNA into plant cells allows for a transient presence of the protein or enzyme that can be useful for biochemical analysis or genome modifications. This may be of particular interest for genome editing, because it can avoid DNA (transgene) integration into the genome and generate precisely modified "nontransgenic" plants. In this work, we explore direct protein delivery to plant cells using mesoporous silica nanoparticles (MSNs) as carriers to deliver Cre recombinase protein into maize (Zea mays) cells. Cre protein was loaded inside the pores of gold-plated MSNs, and these particles were delivered by the biolistic method to plant cells harboring loxP sites flanking a selection gene and a reporter gene. Cre protein was released inside the cell, leading to recombination of the loxP sites and elimination of both genes. Visual selection was used to select recombination events from which fertile plants were regenerated. Up to 20% of bombarded embryos produced calli with the recombined loxP sites under our experimental conditions. This direct and reproducible technology offers an alternative for DNA-free genome-editing technologies in which MSNs can be tailored to accommodate the desired enzyme and to reach the desired tissue through the biolistic method.

Atrazine Degradation Using Immobilized Triazine Hydrolase from Arthrobacter aurescens TC1 in Mesoporous Silica Nanomaterials.

Triazine hydrolase fromArthrobacter aurescens TC1 (TrzN) was successfully immobilized on mesoporous silica nanomaterials (MSNs) for the first time. For both nonfunctionalized MSNs and MSNs functionalized with Zn(II), three pore sizes were evaluated for their ability to immobilize wild-type TrzN: Mobile composition of matter no. 41 (small, 3 nm pores), mesoporous silica nanoparticle material with 10 nm pore diameter (MSN-10) (medium, 6-12 nm pores), and pore-expanded MSN-10 (large, 15-30 nm pores). Of these six TrzN:MSN biomaterials, it was shown that TrzN:MSN-10 was the most active (3.8 ± 0.4 × 10-5 U/mg) toward the hydrolysis of a 50 µM atrazine solution at 25 °C. The TrzN:MSN-10 biomaterial was then coated in chitosan (TrzN:MSN-10:Chit) as chitosan has been shown to increase stability in extreme conditions such as low/high pH, heat shock, and the presence of organic solvents. TrzN:MSN-10:Chit was shown to be a superior TrzN biomaterial to TrzN:MSN-10 as it exhibited higher activity under all storage conditions, in the presence of 20% MeOH, at low and high pH values, and at elevated temperatures up to 80 °C. Finally, the TrzN:MSN-10:Chit biomaterial was shown to be fully active in river water, which establishes it as a functional biomaterial under actual field conditions. A combination of these data indicate that the TrzN:MSN-10:Chit biomaterial exhibited the best overall catalytic profile making it a promising biocatalyst for the bioremediation of atrazine.

Parameters affecting the efficient delivery of mesoporous silica nanoparticle materials and gold nanorods into plant tissues by the biolistic method.

Applying nanotechnology to plant science requires efficient systems for the delivery of nanoparticles (NPs) to plant cells and tissues. The presence of a cell wall in plant cells makes it challenging to extend the NP delivery methods available for animal research. In this work, research is presented which establishes an efficient NP delivery system for plant tissues using the biolistic method. It is shown that the biolistic delivery of mesoporous silica nanoparticle (MSN) materials can be improved by increasing the density of MSNs through gold plating. Additionally, a DNA-coating protocol is used based on calcium chloride and spermidine for MSN and gold nanorods to enhance the NP-mediated DNA delivery. Furthermore, the drastic improvement of NP delivery is demonstrated when the particles are combined with 0.6 µm gold particles during bombardment. The methodology described provides a system for the efficient delivery of NPs into plant cells using the biolistic method.

Ligand conformation dictates membrane and endosomal trafficking of arginine-glycine-aspartate (RGD)-functionalized mesoporous silica nanoparticles.

Recent breakthrough research on mesoporous silica nanoparticle (MSN) materials has illustrated their significant potential in biological applications due to their excellent drug delivery and endocytotic behavior. We set out to determine if MSN, covalently functionalized with conformation specific bioactive molecules (either linear or cyclic RGD ligands), behave towards mammalian cells in a similar manner as the free ligands. We discovered that RGD immobilized on the MSN surface did not influence the integrity of the porous matrix and improved the endocytosis efficiency of the MSN materials. Through competition experiments with free RGD ligands, we also discovered a conformation specific receptor-integrin association. The interaction between RGD immobilized on the MSN surface and integrins plays an important role in endosome trafficking, specifically dictating the kinetics of endosomal escape. Thus, covalent functionalization of biomolecules on MSN assists in the design of a system for controlling the interface with cancer cells.

Chemically reducible lipid bilayer coated mesoporous silica nanoparticles demonstrating controlled release and HeLa and normal mouse liver cell biocompatibility and cellular internalization.

A controlled release system composed of mesoporous silica nanoparticles with covalently bound dipalmitoyl moieties supporting phosphorylated lipids has been successfully synthesized and characterized. This MSN system demonstrates controlled release of fluorescein molecules under disulfide reducing conditions. Flow cytometry analyses confirm increased biocompatibility of the resulting lipid bilayer MSNs (LB-MSNs) from nonfunctional MSNs. Fluorescently labeled LB-MSNs are examined via confocal fluorescent microscopy ex vivo and were found to enter both normal and cancer cell lines. The LB-MSNs presented here have potential to be used as rapid and diverse functionalized, stable liposome analogues for drug delivery.

Breaking the fibrinolytic speed limit with microwheel co-delivery of tissue plasminogen activator and plasminogen.

BACKGROUND: To reestablish blood flow in vessels occluded by clots, tissue plasminogen activator (tPA) can be used; however, its efficacy is limited by transport to and into a clot and by the depletion of its substrate, plasminogen. OBJECTIVES: To overcome these rate limitations, a platform was designed to co-deliver tPA and plasminogen based on microwheels (µwheels), wheel-like assemblies of superparamagnetic colloidal beads that roll along surfaces at high speeds. METHODS: The biochemical speed limit was determined by measuring fibrinolysis of plasma clots at varying concentrations of tPA (10-800 nM) and plasminogen (1-6 µM). Biotinylated magnetic mesoporous silica nanoparticles were synthesized and bound to streptavidin-coated superparamagnetic beads to make studded beads. Studded beads were loaded with plasminogen and tPA was immobilized on their surface. Plasminogen release and tPA activity were measured on the studded beads. Studded beads were assembled into µwheels with rotating magnetic fields and fibrinolysis of plasma clots was measured in a microfluidic device. RESULTS: The biochemical speed limit for plasma clots was ~15 µm/min. Plasminogen-loaded, tPA-immobilized µwheels lyse plasma clots at rates comparableto the biochemical speed limit. With the addition of a corkscrew motion, µwheels penetrate clots, thereby exceeding the biochemical speed limit (~20 µm/min) and achieving lysis rates 40-fold higher than 50 nM tPA. CONCLUSIONS: Co-delivery of an immobilized enzyme and its substrate via a microbot capable of mechanical work has the potential to target and rapidly lyse clots that are inaccessible by mechanical thrombectomy devices or recalcitrant to systemic tPA delivery.

Luciferase and luciferin co-immobilized mesoporous silica nanoparticle materials for intracellular biocatalysis.

We report a gold nanoparticle (AuNP)-capped mesoporous silica nanoparticle (Au-MSN) platform for intracellular codelivery of an enzyme and a substrate with retention of bioactivity. As a proof-of-concept demonstration, Au-MSNs are shown to release luciferin from the interior pores of MSN upon AuNP uncapping in response to disulfide-reducing antioxidants and codeliver bioactive luciferase from the PEGylated exterior surface of Au-MSN to Hela cells. The effectiveness of luciferase-catalyzed luciferin oxidation and luminescence emission in the presence of intracellular ATP was measured by a luminometer. Overall, the chemical tailorability of the Au-MSN platform to retain enzyme bioactivity, the ability to codeliver enzyme and substrate, and the potential for imaging tumor growth and metastasis afforded by intracellular ATP- and glutathione-dependent bioluminescence make this platform appealing for intracellular controlled catalysis and tumor imaging.

Mesoporous silica nanoparticles for intracellular controlled drug delivery.

The application of nanotechnology in the field of drug delivery has attracted much attention in the latest decades. Recent breakthroughs on the morphology control and surface functionalization of inorganic-based delivery vehicles, such as mesoporous silica nanoparticles (MSNs), have brought new possibilities to this burgeoning area of research. The ability to functionalize the surface of mesoporous-silica-based nanocarriers with stimuli-responsive groups, nanoparticles, polymers, and proteins that work as caps and gatekeepers for controlled release of various cargos is just one of the exciting results reported in the literature that highlights MSNs as a promising platform for various biotechnological and biomedical applications. This review focuses on the most recent progresses in the application of MSNs for intracellular drug delivery. The latest research on the pathways of entry into live mammalian and plant cells together with intracellular trafficking are described. One of the main areas of interest in this field is the development of site-specific drug delivery vehicles; the contribution of MSNs toward this topic is also summarized. In addition, the current research progress on the biocompatibility of this material in vitro and in vivo is discussed. Finally, the latest breakthroughs for intracellular controlled drug release using stimuli-responsive mesoporous-silica-based systems are described.

Mesoporous silica nanoparticle-based double drug delivery system for glucose-responsive controlled release of insulin and cyclic AMP.

A boronic acid-functionalized mesoporous silica nanoparticle-based drug delivery system (BA-MSN) for glucose-responsive controlled release of both insulin and cyclic adenosine monophosphate (cAMP) was synthesized. Fluorescein isothiocyanate-labeled, gluconic acid-modified insulin (FITC-G-Ins) proteins were immobilized on the exterior surface of BA-MSN and also served as caps to encapsulate cAMP molecules inside the mesopores of BA-MSN. The release of both G-Ins and cAMP was triggered by the introduction of saccharides. The selectivity of FITC-G-Ins release toward a series of carbohydrate triggers was determined to be fructose > glucose > other saccharides. The unique feature of this double-release system is that the decrease of FITC-G-Ins release with cycles can be balanced by the release of cAMP from mesopores of MSN, which is regulated by the gatekeeper effect of FITC-G-Ins. In vitro controlled release of cAMP was studied at two pH conditions (pH 7.4 and 8.5). Furthermore, the cytotoxicity of cAMP-loaded G-Ins-MSN with four different cell lines was investigated by cell viability and proliferation studies. The cellular uptake properties of cAMP-loaded FITC-BA-MSN with and without G-Ins capping were investigated by flow cytometry and fluorescence confocal microscopy. We envision that this glucose-responsive MSN-based double-release system could lead to a new generation of self-regulated insulin-releasing devices.

Catalytic reactions of carbene precursors on bulk gold metal.

Bulk gold metal powder, consisting of particles (5-50 microm) much larger than nanoparticles, catalyzes the coupling of carbenes generated from diazoalkanes (R(2)C=N(2)) and 3,3-diphenylcyclopropene (DPCP) to form olefins. It also catalyzes cyclopropanation reactions of these carbene precursors with styrenes. The catalytic activity of the gold powder depends on the nature of the gold particles, as determined by TEM and SEM studies. The reactions can be understood in terms of mechanisms that involve the generation of carbene R(2)C: intermediates adsorbed on the gold surface.

Microcolumn lanthanide separation using bis-(2-ethylhexyl) phosphoric acid functionalized ordered mesoporous carbon materials.

Adjacent lanthanides are among the most challenging elements to separate, to the extent that current separations materials would benefit from transformative improvement. Ordered mesoporous carbon (OMC) materials are excellent candidates, owing to their small mesh size and uniform morphology. Herein, OMC materials were physisorbed with bis-(2-ethylhexyl) phosphoric acid (HDEHP) and sorption of Eu3+ was investigated under static and dynamic conditions. The HDEHP-OMC materials displayed higher distribution coefficients and loading capacities than current state-of-the-art materials. Using a small, unpressurized column, a separation between Eu3+ and Nd3+ was achieved. Based on these experimental results, HDEHP-OMC have shown potential as a solid phase sorbent for chromatographic, intragroup, lanthanide separations.

Endocytosis of a single mesoporous silica nanoparticle into a human lung cancer cell observed by differential interference contrast microscopy.

The unique structural features of mesoporous silica nanoparticles (MSN) have made them very useful in biological applications, such as gene therapy and drug delivery. Flow cytometry, confocal microscopy, and electron microscopy have been used for observing the endocytosis of MSN. However, flow cytometry cannot directly observe the process of endocytosis. Confocal microscopy requires fluorescence labeling of the cells. Electron microscopy can only utilize fixed cells. In the present work, we demonstrate for the first time that differential interference contrast (DIC) microscopy can be used to observe the entire endocytosis process of MSN into living human lung cancer cells (A549) without fluorescence staining. There are three physical observables that characterize the locations of MSN and the stages of the endocytosis process: motion, shape, and vertical position. When it was outside the cell, the MSN underwent significant Brownian motion in the cell growth medium. When it was trapped on the cell membrane, the motion of the MSN was greatly limited. After the MSN had entered the cell, it resumed motion at a much slower speed because the cytoplasm is more viscous than the cell growth medium and the cellular cytoskeleton networks act as obstacles. Moreover, there were shape changes around the MSN due to the formation of a vesicle after the MSN had been trapped on the cell membrane and prior to entry into the cell. Finally, by coupling a motorized vertical stage to the DIC microscope, we recorded the location of the MSN in three dimensions. Such accurate 3D particle tracking ability in living cells is essential for studies of selectively targeted drug delivery based on endocytosis.

Palladium Intercalated into the Walls of Mesoporous Silica as Robust and Regenerable Catalysts for Hydrodeoxygenation of Phenolic Compounds.

Nanostructured noble-metal catalysts traditionally suffer from sintering under high operating temperatures, leading to durability issues and process limitations. The encapsulation of nanostructured catalysts to prevent loss of activity through thermal sintering, while maintaining accessibility of active sites, remains a great challenge in the catalysis community. Here, we report a robust and regenerable palladium-based catalyst, wherein palladium particles are intercalated into the three-dimensional framework of SBA-15-type mesoporous silica. The encapsulated Pd active sites remain catalytically active as demonstrated in high-temperature/pressure phenol hydrodeoxygenation reactions. The confinement of Pd particles in the walls of SBA-15 prevents particle sintering at high temperatures. Moreover, a partially deactivated catalyst containing intercalated particles is regenerated almost completely even after several reaction cycles. In contrast, Pd particles, which are not encapsulated within the SBA-15 framework, sinter and do not recover prior activity after a regeneration procedure.

Mesoporous silica nanoparticle based controlled release, drug delivery, and biosensor systems.

Recent advancements in controlling the surface properties and particle morphology of the structurally defined mesoporous silica materials with high surface area (>700 m(2) g(-1)) and pore volume (>1 cm(3) g(-1)) have significantly enhanced their biocompatibility. Various methods have been developed for the functionalization of both the internal pore and exterior particle surfaces of these silicates with a tunable pore diameter ranging from 2 to 30 nm and a narrow pore size distribution. Herein, we review the recent research progress on the design of functional mesoporous silica materials for stimuli-responsive controlled release delivery of pharmaceutical drugs, genes, and other chemicals. Furthermore, the recent breakthroughs in utilizing these nanoscale porous materials as sensors for selective detections of various neurotransmitters and biological molecules are summarized.

Nanoparticle-Mediated Recombinase Delivery into Maize.

We describe a non-DNA-based system for delivering Cre recombinase protein into maize tissue using gold-plated mesoporous silica nanoparticle (Au-MSN). Cre protein is first loaded into the pores of Au-MSNs and then delivered using the biolistic method to immature embryos of a maize line (Lox-corn), which harbors loxP sites flanking a selection and a reporter gene. The release of the Cre recombinase protein inside the plant cell leads to recombination at the loxP sites, eliminating both genes. Visual screening is used to identify recombination events, which can be regenerated to mature and fertile plants. Using the experimental procedures and conditions described here, as high as 20% of bombarded embryos can produce regenerable recombinant callus events. This nanomaterial-mediated, DNA-free methodology has potential to become an effective tool for plant genome editing.