The aim of this in vitro study is to prepare and characterize drug free and pentamidine loaded chitosan glutamate coated niosomes for intranasal drug delivery to reach the brain through intranasal delivery. Mucoadhesive properties and stability testing in various environments were evaluated to examine the potential of these formulations to be effective drug delivery vehicles for intranasal delivery to the brain. Samples were prepared using thin film hydration method. Changes in size and ζ-potential of coated and uncoated niosomes with and without loading of pentamidine in various conditions were assessed by dynamic light scattering (DLS), while size and morphology were also studied by atomic force microscopy (AFM). Bilayer properties and mucoadhesive behavior were investigated by fluorescence studies and DLS analyses, respectively. Changes in vesicle size and ζ-potential values were shown after addition of chitosan glutamate to niosomes, and when in contact with mucin solution. In particular, interactions with mucin were observed in both drug free and pentamidine loaded niosomes regardless of the presence of the coating. The characteristics of the proposed systems, such as pentamidine entrapment and mucin interaction, show promising results to deliver pentamidine or other possible drugs to the brain via nasal administration.
Silver nanoparticles (AgNPs) are widely used as antibacterial agents and anticancer drugs, but often their low stability limits their mass production and broad applications. The use of niosomes as a carrier to protect and envelop AgNPs gives a new perspective to solve these problems. In this study, AgNPs were functionalized with sodium 3-mercapto-1-propanesulfonate (3MPS) to induce hydrophilic behavior, improving loading in Tween 20 and Span 20 niosomes (NioTw20 and NioSp20, respectively). Entrapment efficiency was evaluated by UV analyses and is around 1–4%. Dimensions were investigated by means of dynamic light scattering (DLS) ( = 140 ± 4 nm and = 251 ± 1 nm respectively for NioTw20 + AgNPs and NioSp20 + AgNPs) and were compared with those by atomic force microscopy (AFM) and small angle X ray scattering (SAXS) analyses. Stability was assessed in water up to 90 days, and both in bovine serum and human serum for up to 8 h. In order to characterize the local structure of niosomes, SAXS measurements have been performed on Tween 20 and Span 20 empty niosomes and loaded with AgNPs. The release profiles of hydrophilic probe calcein and lipophilic probe Nile Red were performed in HEPES buffer and in human serum. All these features contribute to conclude that the two systems, NioTw20 + AgNPs and NioSp20 + AgNPs, are suitable and promising in the field of biological applications.
The development of high spatial resolution and element sensitive magnetic characterization techniques to quantitatively measure magnetic parameters of individual nanoparticles (NPs) and deeply understand and tune their magnetic properties is a hot topic in nanomagnetism. Magnetic force microscopy (MFM), thanks to its high lateral resolution, appears as a promising technique for the magnetic characterization of single nano-sized materials although it is still limited by some drawbacks, especially by the presence of electrostatic artifacts. Recently, these limitations have been overcome by the development of a particular MFM based technique called controlled magnetization – MFM (CM-MFM) allowing, in principle, a quantifiable correlation between the measured magnetic signal and the magnetization of the object under investigation. Here we propose an experimental procedure, based on the use of CM-MFM technique, to measure the magnetization curve of single magnetic NPs individuating their saturation magnetization, magnetic field, and coercivity. We measured, for the first time, the magnetization curves of individual Fe3O4 nanoparticles with diameters in the range of 18–32 nm by using a MFM instrument. Results are in very good agreement with the quantitative data obtained by SQUID analysis on a macroscopic sample, showing the high potential of the technique in the field of nanomagnetometry.
The development of magnetic nanoparticles for biomedical applications requires a detailed characterization of their magnetic properties, with relation not only to their chemical structure, but also their morphology and size. Magnetic force microscopy (MFM), thanks to its nanometric lateral resolution and its capability to detect weak magnetic fields, appears as a powerful tool for the characterization of the magnetic properties of single nanoparticles, together with their morphological characteristics. Nevertheless, the application of MFM to the quantitative measurements of magnetic properties at the nanoscale is still an open issue because of a certain incongruence between experimental data and existing theoretical models of the tip-sample magnetic interactions. In this work, MFM data acquired on different magnetic nanoparticles in different experimental conditions (magnetized and not magnetized probes, out-of-field and in-field measurements) are analyzed, with the aim of individuating the possible phenomena affecting MFM measurements. These include topography-induced artifacts resulting from the tip-sample capacitive coupling, which we propose here for the first time. In case of measurements performed in presence of an external magnetic field, much more intense MFM signals were detected as it produces the saturation of the magnetization of the nanoparticles, which is not completely obtained by the sole stray field produced by the tip. Nevertheless, even in in-field measurements, the results evidenced the presence of significant electrostatic effects in MFM images, which, therefore, appear as an important factor to be taken into account for the quantitative interpretation of MFM data.
Despite the potential of small‐scale pillars of black titanium (bTi) for killing the bacteria and directing the fate of stem cells, not much is known about the effects of the pillars’ design parameters on their biological properties. Here, three distinct bTi surfaces are designed and fabricated through dry etching of the titanium, each featuring different pillar designs. The interactions of the surfaces with MC3T3‐E1 preosteoblast cells and Staphylococcus aureus bacteria are then investigated. Pillars with different heights and spatial organizations differently influence the morphological characteristics of the cells, including their spreading area, aspect ratio, nucleus area, and cytoskeletal organization. The preferential formation of focal adhesions (FAs) and their size variations also depend on the type of topography. When the pillars are neither fully separated nor extremely tall, the colocalization of actin fibers and FAs as well as an enhanced matrix mineralization are observed. However, the killing efficiency of these pillars against the bacteria is not as high as that of fully separated and tall pillars. This study provides a new perspective on the dual‐functionality of bTi surfaces and elucidates how the surface design and fabrication parameters can be used to achieve a surface topography with balanced bactericidal and osteogenic properties.
Physical patterns represent potential surface cues for promoting osteogenic differentiation of stem cells and improving osseointegration of orthopedic implants. Understanding the early cell–surface interactions and their effects on late cellular functions is essential for a rational design of such topographies, yet still elusive. In this work, fluidic force microscopy (FluidFM) and atomic force microscopy (AFM) combined with optical and electron microscopy are used to quantitatively investigate the interaction of preosteoblasts with 3D‐printed patterns after 4 and 24 h of culture. The patterns consist of pillars with the same diameter (200 nm) and interspace (700 nm) but distinct heights (500 and 1000 nm) and osteogenic properties. FluidFM reveals a higher cell adhesion strength after 24 h of culture on the taller pillars (32 ± 7 kPa versus 21.5 ± 12.5 kPa). This is associated with attachment of cells partly on the sidewalls of these pillars, thus requiring larger normal forces for detachment. Furthermore, the higher resistance to shear forces observed for these cells indicates an enhanced anchorage and can be related to the persistence and stability of lamellipodia. The study explains the differential cell adhesion behavior induced by different pillar heights, enabling advancements in the rational design of osteogenic patterns.