Teemu Heikkilä

In vitro cytotoxicity of porous silicon microparticles: Effect of the particle concentration, surface chemistry and size

We report here the in vitro cytotoxicity of mesoporous silicon (PSi) microparticles on the Caco-2 cells as a function of particle size fractions (1.2-75 microm), particle concentration (0.2-4 mg ml(-1)) and incubation times (3, 11 and 24 h). The particle size (smaller PSi particles showed higher cytotoxicity) and the surface chemistry treatment of the PSi microparticles were considered to be the key factors regarding the toxicity aspects. These effects were significant after the 11 and 24 h exposure times, and were explained by cell-particle interactions involving mitochondrial disruption resulting from ATP depletion and reactive oxygen species production induced by the PSi surface. These events further induced an increase in cell apoptosis and consequent cell damage and cell death in a dose-dependent manner and as a function of the PSi particle size. These effects were, however, less pronounced with thermally oxidized PSi particles. Under the experimental conditions tested and at particle sizes >25 microm, the non-toxic threshold concentration for thermally hydrocarbonized and carbonized PSi particles was <2 mg ml(-1), and for thermally oxidized PSi microparticles was <4 mg ml(-1).

Evaluation of Mesoporous TCPSi, MCM-41, SBA-15, and TUD-1 Materials as API Carriers for Oral Drug Delivery

The feasibility of four mesoporous materials composed of biocompatible Si (TCPSi) or SiO2 (MCM-41, SBA-15, and TUD-1) were evaluated for oral drug delivery applications. The main focus was to study the effect of the materials different pore systems (unidirectional/2D/3D) and their pore diameters, pore size distributions, pore volumes on the maximal drug load capacity, and release profiles of a loaded active pharmaceutical ingredient. Ibuprofen was used as the model drug. The total pore volume of the mesoporous solid was the main factor limiting the maximum drug load capacity, with SBA-15 reaching a very high drug load of 1:1 in weight due to its high pore volume. Dissolution experiments were performed in HBSS buffers of pH 5.5, 6.8, and 7.4 to mimic the conditions in the small intestine. At pH 5.5 the dissolution rate of ibuprofen released from the mesoporous carriers was significantly faster compared with the standard bulk ibuprofen (86–63% versus 25% released at 45 min), with the fastest release observed from the 3D pore network of TUD-1 carrier. The utilization of mesoporous carriers diminished the pH dependency of ibuprofen dissolution (pKa = 4.42), providing an interesting prospect for the formulation of poorly soluble drug compounds.

Drug Delivery Formulations of Ordered and Nonordered Mesoporous Silica: Comparison of Three Drug Loading Methods

A poorly soluble model drug, indomethacin (IMC), was loaded into two types of silica particles using three different loading methods. The loading efficiency and the extent/rate of drug release were evaluated. Widely used equipment in pharmaceutical laboratories, rotavapor and fluid bed, were used in the loading. The porous materials used were ordered mesoporous silica MCM-41 and nonordered silica gel Syloid 244 FP EU. The materials differ both in their pore properties and particle sizes. Tablets were successfully compressed from the IMC-loaded particles. Mechanical stability of the porous structures was studied with XRPD and nitrogen sorption after tableting and drug release was evaluated at pH 5.5 before and after tableting. The release of the poorly soluble IMC was faster from the Syloid than from the MCM-41, presumably due to the larger pore size and smaller particle size. Loading of IMC into the MCM-41 microparticles improved the drug dissolution, and blending the microparticles with pharmaceutical excipients improved the IMC release even further. The fast release was also maintained after tableting. Loading of IMC into the Syloid particles alone was sufficient to produce similar IMC release profiles, as in the case of MCM-41 with the excipients.

Ruthenium-modified MCM-41 mesoporous molecular sieve and Y zeolite catalysts for selective hydrogenation of cinnamaldehyde

Abstract Ru-modified MCM-41 mesoporous molecular sieve and Y zeolite catalysts were synthesised and characterised using XRD, electrochemical voltammetry, ESCA (XPS), nitrogen adsorption and H2-TPD. Selective liquid-phase hydrogenation of cinnamaldehyde to cinnamyl alcohol was investigated over these catalysts and compared to hydrogenation on a commercial Ru/C catalyst. The effect of support (MCM-41, Y and C), solvent (cyclohexane, hexane and 2-propanol) and catalyst pre-treatment (calcination) on the conversion of cinnamaldehyde and selectivity to cinnamyl alcohol was studied. The zeolite structure, pore size, acidity, catalyst pre-treatment as well as the solvent influenced the activity and selectivity. Non-calcined Ru/Y exhibited the highest selectivity to cinnamyl alcohol. The activity of Ru/Y was highest with 2-propanol as a solvent.

¹⁸F-labeled modified porous silicon particles for investigation of drug delivery carrier distribution in vivo with positron emission tomography.

Because of its biocompatibility and ability to accommodate a variety of payloads from poorly soluble drugs to biomolecules, porous silicon (PSi) is a lucrative material for the development of carriers for particle-mediated drug delivery. We report a successful direct one-step (18)F-radiolabeling of three types of PSi microparticles, thermally hydrocarbonized THCPSi, thermally oxidized TOPSi, and thermally carbonized TCPSi for the investigation of their biodistribution in vivo with positron emission tomography as part of their evaluation as carriers for particle-mediated drug delivery. FTIR and XPS characterization of the PSi materials after carrier-added (18)F/(19)F-radiolabeling reveals that depending on the material the (18)F-labeling is likely to be accomplished either by substitution for surface silyl hydrogen or silyl fluoride or by nucleophilic attack of (18)F(-) to Si-O-Si bridges. With the selected (18)F-radiolabeling method, good to excellent in vitro radiolabel stability in simulated gastric and intestinal fluids and in plasma is achieved for all the particle types studied. Finally, a preliminary evaluation of (18)F-THCPSi microparticle biodistribution in the rat gastrointestinal tract after oral administration is reported, illustrating the utility of using (18)F-radiolabeled PSi as imaging probes for PSi-based drug delivery carrier distribution in vivo.

Cytotoxicity study of ordered mesoporous silica MCM-41 and SBA-15 microparticles on Caco-2 cells

Cytotoxicity of ordered mesoporous silica MCM-41 and SBA-15 microparticles (fractions between 1 and 160 microm) was determined in vitro on undifferentiated human colon carcinoma (Caco-2) cell line, considering the feasibility of using these silica-based materials in oral drug formulations. The cellular endpoints employed for assessing the effects of the MCM-41 and SBA-15 microparticles on Caco-2 were: (1) cell membrane integrity by monitoring live-cell protease activity (AFC) and by employing the flow cytometry method; (2) metabolic activity by monitoring total ATP content via luminescence assay; (3) activity of apoptotic effectors by caspase-3/7 activity assay. The generation of reactive oxygen species (ROS) was also followed, specifically the hydrogen peroxide (H(2)O(2)) and the superoxide radical (O(2)(-)). MCM-41 and SBA-15 microparticles caused cytotoxic effects on the Caco-2 cells, at most tested concentrations (0.2-14 mg/ml) and incubation times (3 and 24h). The effects on the cells included weakened cell membrane integrity, diminished cell metabolism and increased apoptotic signalling. The root cause for the cytotoxicity was heightened production of reactive oxygen species (ROS), especially the formation of the superoxide radical O(2)(-) already after 3h incubation with threshold dose 1mg/ml, apparently overwhelming the antioxidant defences and causing mitochondrial dysfunction, hence increasing the apoptotic signalling.

Physicochemical stability of high indomethacin payload ordered mesoporous silica MCM-41 and SBA-15 microparticles

Stability of high indomethacin (IMC) content formulations based on ordered mesoporous silica MCM-41 and SBA-15 materials was studied before and after a 3 month storage in stressed conditions (30°C/56% RH). Overall, the physical stability of the samples was found satisfactory after the storage. However, some issues with the chemical stability were noted, especially with the MCM-41 based samples. The stability issues were evident from the decreased HPLC loading degrees of the drug after stressing as well as from the observed extra peaks in the HPLC chromatograms of the drug in the stressed samples. Drug release from the mesoporous formulations before stressing was rapid at pH 1.2 in comparison to bulk crystalline IMC. The release profiles also remained similar after stressing. Even faster and close to complete IMC release was achieved when the pH was raised from 1.2 to 6.8. To our knowledge, this is the first report of chemical stability issues of drugs in mesoporous silica drug formulations. The present results encourage further study of the factors affecting the chemical stability of drugs in mesoporous silica MCM-41 and SBA-15 formulations in order to realize their potential in oral drug delivery.

Formation of Furfural in Catalytic Transformation of Levoglucosan over Mesoporous Materials

Catalytic transformations of levoglucosan (1‐6‐anhydro‐β‐D‐glucopyranose) and furfural were carried out in a fixed‐bed reactor at 573 K over mesoporous materials. Proton forms of MCM‐41, MCM‐48, SBA‐15, and platinum form of MCM‐48 catalysts were tested in the reaction, whereas H‐Beta and quartz sand were used as reference materials. The yield of the transformation products was substantially influenced by the catalyst structures. Oxygenated species were the main liquid products, consisting mainly of aldehydes and furfural. The formation of furfural was the highest over MCM‐41 catalyst followed by SBA‐15, MCM‐48, and H‐Beta catalyst. All catalysts were to some extent deactivated due to coke formation. However, it was possible to successfully regenerate the spent catalysts without changing the structure.

Cu-H-MCM-41, H-MCM-41 and Na-MCM-41 mesoporous molecular sieve catalysts for isomerization of 1-butene to isobutene

Cu–H-MCM-41, H-MCM-41 and Na-MCM-41 mesoporous molecular sieve catalysts were synthesized, characterized and investigated in the isomerization of 1-butene to isobutene. Introduction of copper in MCM-41 was found to play a positive role in enhancing the conversion of 1-butene and yield of isobutene and Cu–H-MCM-41 exhibited higher conversion of 1-butene and yield to isobutene than H-MCM-41 and Na-MCM-41 catalysts. The pretreatments of Cu–H-MCM-41 catalyst with synthetic air or hydrogen were observed to influence the 1-butene conversion, yield of isobutene and selectivity to isobutene. Pre-treated with the synthetic air the Cu–H-MCM-41-Ox catalyst exhibited higher conversion of 1-butene, yield of isobutene and selectivity to isobutene than hydrogen pre-treated Cu–H-MCM-41-Red. The reason for such a behavior of Cu–H-MCM-41-Red is the reduction of copper species to metallic form. The X-ray powder diffraction pattern of Cu–H-MCM-41-Red exhibited a peak attributed to the reduction of Cu2+ to Cu0. FTIR spectra of adsorbed pyridine showed the presence of Brønsted and Lewis acid sites in the H-MCM-41, Na-MCM-41 and Cu–H-MCM-41 catalysts.

Kinetics of lactose and rhamnose oxidation over supported metal catalysts

Several mono- and bimetallic Pd, Pt, Rh and Ru supported on alumina and active carbon catalysts were characterized by CO chemisorption, nitrogen adsorption, XPS and XRD and acidity titrations were performed for active carbon supported catalysts. These catalysts were tested in oxidation of two sugars, namely lactose and rhamnose, at 60 °C and at 70 °C under slightly alkaline conditions (pH 8) with molecular oxygen. The results revealed that there is an optimum metal particle size in a range of 3-10 nm giving the highest initial TOFs for both oxidations. Furthermore, the catalytic activities and conversions were related to other catalyst properties, such as the type and amount of promoters and the presence of different phases. In situ catalyst potential measurements revealed that there is an inverse correlation between the increase of catalyst potential as a function of sugar conversion and the catalyst activity after prolonged reaction times. This method is a valuable tool for in situ characterization of catalysts correlating well with their activities.