Monodispersed Bioactive Glass Nanoparticles Enhance the Osteogenic Differentiation of Adipose‐Derived Stem Cells through Activating TGF‐Beta/Smad3 Signaling Pathway

It is important to understand the interaction mechanisms between nanomaterials and adipose‐derived stem cells for biomedical application. Nanoscale bioactive glass has positive effects on guiding osteoblasts differentiation and bone regeneration. However, the effects and molecular mechanism of monodispersed bioactive glass nanoparticles on the osteogenic differentiation of adipose‐derived stem cells are still not clear up to now. In this study, the effects and underlying molecular mechanism of monodispersed bioactive glass nanoparticles on the osteogenic differentiation of adipose‐derived stem cells are investigated in minute detail. The results show that nanoparticles (100–200 nm) can be absorbed by stem cells and is distributed in cytoplasm and nucleus. In both culture conditions (normal and osteoinductive), nanoparticles (80 µg mL⁻¹) can significantly enhance the osteogenic differentiation of stem cells through upregulating the alkaline phosphatase activity, osteogenic genes and protein expressions, as well as calcium deposition. Further study suggests that the activation of transforming growth factor‐beta/Smad3 signaling pathway plays an important role in the osteogenic differentiation of adipose‐derived stem cells enhanced by monodispersed nanoparticles. This study may have important implications for better understanding of stem cells fate induced by monodispersed nanoparticles and provide a promising approach toward stem cells‐based bone regeneration.     

Analysis of 4-dimethylaminopyridine (DMAP)-gold nanoparticles behaviour in solution and of their interaction with calf thymus DNA and living cells

4-(Dimethylamino)pyridine-coated gold nanoparticles (DMAP-Au NPs) were synthesized, characterised and their interaction with DNA and living cells was analysed. Concerning the interaction of the DMAP-Au NPs with DNA, absorbance titrations indicate that a non-covalent interaction between DNA and the external surface of the NPs does take place. The binding constant was evaluated to be (2.8 ± 0.8) × 10⁵ M⁻¹. Exposure of cultured cells to NPs revealed a dose-dependent effect on cell proliferation which was increased or reduced in dependence of DMAP-Au NPs concentrations. Subcellular localisation by transmission electron microscopy showed mitochondrial and nuclear localisations of NPs, thus suggesting their direct involvement in the mitochondrial alterations observed and a possible direct interaction with cell DNA. These findings clearly indicate that DMAP-Au NPs can strongly interact with living cells and confirm the importance of systematic evaluations of NPs properties, also in the perspective of their arising diagnostic and therapeutic applications.     

Electro-acupuncture promotes survival, differentiation of the bone marrow mesenchymal stem cells as well as functional recovery in the spinal cord-transected rats

Background  

Bone marrow mesenchymal stem cells (MSCs) are one of the potential tools for treatment of the spinal cord injury; however, the survival and differentiation of MSCs in an injured spinal cord still need to be improved. In the present study, we investigated whether Governor Vessel electro-acupuncture (EA) could efficiently promote bone marrow mesenchymal stem cells (MSCs) survival and differentiation, axonal regeneration and finally, functional recovery in the transected spinal cord.     

In situ Raman spectroscopic monitoring of hydroxyapatite as human mesenchymal stem cells differentiate into osteoblasts

The differentiation of stem cells into specific cell types is playing an essential role in the development of stem cell therapy, tissue engineering, and regenerative medicine. In this research, Raman microspectroscopy was applied to monitor the development of hydroxyapatite [HA, Ca₅ (PO₄)₃ (OH)], which is associated with the differentiation of the human mesenchymal stem cells (hMSCs) into osteoblasts. Raman spectra exhibited dramatic changes in the HA region, 950–970 cm⁻¹, over the period of 7–21 days after the start of differentiation. This work demonstrates the successful application of Raman spectroscopy for monitoring and quantitatively evaluating hMSC differentiation into osteoblasts. Copyright © 2008 John Wiley & Sons, Ltd.     

Manganese–gold nanoparticles as an MRI positive contrast agent in mesenchymal stem cell labeling

We report a straightforward approach to prepare multifunctional manganese–gold nanoparticles by attaching Mn(II) ions onto the surface of 20 nm citrate-capped gold nanoparticles. In vitro MRI measurements made in agarose gel phantoms exhibited high relaxivity (18.26 ± 1.04 mmol⁻¹ s⁻¹). Controlled incubation of the nanoparticles with mesenchymal stem cells (MSCs) was used to study cellular uptake of these particles and this process appeared to be controlled by the size of the nanoparticle aggregates in the extracellular solution. SEM images of live MSCs showed an increased concentration of particles near the cell membrane and a distribution of the size of particles within the cells. Survivability for MSCs in contact with Mn–Au NPs was greater than 97% over the 3-day period and up to the 1 mM Mn used in this study. The high relaxivity and low cell mortality are suggestive of an enhanced positive contrast agent for in vitro or in vivo applications.     

Development of poly(butylene succinate)/calcium phosphate composites for bone engineering

Bone tissue engineering offers the prospect of alternative therapies for clinically relevant skeletal defects. Poly(butylene succinate) (PBSu) is a biodegradable and biocompatible polyester which possesses some unfavorable biomaterial properties. In order to improve this limitation, we developed PBSu/hydroxyapatite (HA) and PBSu/β-tricalcium phosphate (TCP) composites to support the growth and osteogenic differentiation of human mesenchymal stem cells (hMSCs). The results showed that phase separation morphology of the composites were detected in both PBSu/HA and PBSu/TCP films where calcium phosphate (HA and TCP) dispersed thoroughly into PBSu. The addition of either HA or TCP increased the hydrophilicity of the resulting composites. All the materials appeared to be biocompatible and supported in vitro growth and osteoblast differentiation of hMSCs. In conclusion, the currently developed composite materials possess good biocompatibility and allow the growth and osteogenic differentiation of hMSCs in vitro, suggesting their potential application in stem cell-based bone engineering.     

Effect of gold nanoparticle size on acoustic cavitation using chemical dosimetry method

When a liquid is irradiated with high intensities of ultrasound irradiation, acoustic cavitation occurs. Acoustic cavitation generates free radicals from the breakdown of water and other molecules. Cavitation can be fatal to cells and is utilized to destroy cancer tumors. The existence of particles in liquid provides nucleation sites for cavitation bubbles and leads to decrease the ultrasonic intensity threshold needed for cavitation onset. In the present investigation, the effect of gold nanoparticles with appropriate amount and size on the acoustic cavitation activity has been shown by determining hydroxyl radicals in terephthalic acid solutions containing 15, 20, 28 and 35 nm gold nanoparticles sizes by using 1 MHz low level ultrasound. The effect of sonication intensity in hydroxyl radical production was considered.The recorded fluorescence signal in terephthalic acid solutions containing gold nanoparticles was considerably higher than the terephthalic acid solutions without gold nanoparticles at different intensities of ultrasound irradiation. Also, the results showed that the recorded fluorescence signal intensity in terephthalic acid solution containing finer size of gold nanoparticles was lower than the terephthalic acid solutions containing larger size of gold nanoparticles. Acoustic cavitation in the presence of gold nanoparticles can be used as a way for improving therapeutic effects on the tumors.     

Blood Compatibility of Multilayered Polyelectrolyte Films Containing Immobilized Gold Nanoparticles

Surface material functionalization including layer‐by‐layer (LbL) polyelectrolyte films with incorporated nanoparticles is a growing field with a wide range of biomedical applications: drug reservoirs, medical devices, or tissue engineering. In parallel, gold nanoparticles (AuNPs) can be grafted by drugs and sensitive molecules using simple protocols. This study shows that AuNP behavior is modified when they are entrapped into three partner LbL films in comparison to the colloidal solution. A polycationic (polyallylamine hydrochloride (PAH)) and a polyanionic (polyacrylic acid (PAA)) polymer is used to build films based on three cycles ((PAH/AuNP/PAA)₃). To investigate the interaction with biomolecules and cells, three different films are developed changing the outer layer (either PAH or AuNP or PAA) with the same number of AuNP deposit. The best biocompatibility is observed with a polyacrylic acid outer layer. Due to the high capacity of drug grafting on gold nanoparticles, the results seem promising for the development of nanostructured biomedical devices.     

Differentiation of human bone marrow stem cells into cells with a neural phenotype: diverse effects of two specific treatments

Background  

It has recently been demonstrated that the fate of adult cells is not restricted to their tissues of origin. In particular, it has been shown that bone marrow stem cells can give rise to cells of different tissues, including neural cells, hepatocytes and myocytes, expanding their differentiation potential.     

The influence of thermal annealing on the structural and magnetic properties of gold nanoparticles

In the past few years ferromagnetic-like behavior has been reported in metal gold nanoparticles coated with diverse organic surfactants. In this work we report on the effect of thermal annealing on the ferromagnetic-like behavior of oleic acid and oleylamine coated gold nanoparticles of about 7 nm size. The magnetic moment of the “as prepared” sample is about 3×10⁻² emu/g and the coercive field is 200 Oe at 10 kOe and 5 K, after the annealing the behavior changes from ferromagnetic-like to paramagnetic and the magnetization at 10 kOe decreases at a factor of 10. These results are compared with those obtained for oleylamine coated gold nanoparticles, which are diamagnetic at room temperature.     

Seed-mediated shape evolution of gold nanomaterials: from spherical nanoparticles to polycrystalline nanochains and single-crystalline nanowires

We studied the kinetics of the reduction of a gold precursor (HAuCl₄) and the effect of the molar ratio (R) of sodium citrate, which was introduced from a seed solution, and the gold precursor on the shape evolution of gold nanomaterials in the presence of preformed 13 nm gold nanoparticles as seeds. The reduction of the gold precursor by sodium citrate was accelerated due to the presence of gold seeds. Nearly single-crystalline gold nanowires were formed at a very low R value (R = 0.16) in the presence of the seeds as a result of the oriented attachment of the growing gold nanoparticles. At a higher R value (R = 0.33), gold nanochains were formed due to the non-oriented attachment of gold nanoparticles. At a much higher R value (R = 1.32), only larger spherical gold nanoparticles grown from the seeds were found. In the absence of gold seeds, no single-crystalline nanowires were formed at the same R value. Our results indicate that the formation of the 1D nanostructures (nanochains and nanowires) at low R values is due to the attachment of gold nanoparticles along one direction, which is driven by the surface energy reduction, nanoparticle attraction, and dipole–dipole interaction between adjacent nanoparticles.     

Room-temperature synthesis of gold nanoparticles and nanoplates using Shewanella algae cell extract

Biosynthesis of spherical gold nanoparticles and gold nanoplates was achieved at room temperature and pH 2.8 when cell extract from the metal-reducing bacterium Shewanella algae was used as both a reducing and shape-controlling agent. Cell extract, prepared by sonicating a suspension of S. algae cells, was capable of reducing 1 mol/m³ aqueous AuCl₄ ⁻ ions into elemental gold within 10 min when H₂ gas was provided as an electron donor. The time interval lapsed since the beginning of the bioreductive reaction was found to be an important factor in controlling the morphology of biogenic gold nanoparticles. After 1 h, there was a large population of well-dispersed, spherical gold nanoparticles with a mean size of 9.6 nm. Gold nanoplates with an edge length of 100 nm appeared after 6 h, and 60% of the total nanoparticle population was due to gold nanoplates with an edge length of 100–200 nm after 24 h. The yield of gold nanoplates prepared with S. algae extract was four times higher than that prepared with resting cells of S. algae. The resulting biogenic gold nanoparticle suspensions showed a large variation in color, ranging from pale pink to purple due to changes in nanoparticle morphology.     

Extracellular biosynthesis of monodispersed gold nanoparticles by a SAM capping route

Monodispersed gold nanoparticles capped with a self-assembled monolayer of dodecanethiol were biosynthesized extracellularly by an efficient, simple, and environmental friendly procedure, which involved the use of Bacillus megatherium D01 as the reducing agent and the use of dodecanethiol as the capping ligand at 26 °C. The kinetics of gold nanoparticle formation was followed by transmission electron microscope (TEM) and UV-vis spectroscopy. It was shown that reaction time was an important parameter in controlling the morphology of gold nanoparticles. The effect of thiol on the shape, size, and dispersity of gold nanoparticles was also studied. The results showed that the presence of thiol during the biosynthesis could induce the formation of small size gold nanoparticles (<2.5 nm), hold the shape of spherical nanoparticles, and promote the monodispersity of nanoparticles. Through the modulation of reaction time and the use of thiol, monodispersed spherical gold nanoparticles capped with thiol of 1.9 ± 0.8 nm size were formed by using Bacillus megatherium D01.     

Use of a silver ion selective electrode to assess mechanisms responsible for biological effects of silver nanoparticles

For a detailed analysis of the biological effects of silver nanoparticles, discrimination between effects related to the nano-scale size of the particles and effects of released silver ions is required. Silver ions are either present in the initial particle dispersion or released by the nanoparticles over time. The aim of this study is to monitor the free silver ion activity {Ag⁺} in the presence of silver nanoparticles using a silver ion selective electrode. Therefore, silver in the form of silver nanoparticles, 4.2 ± 1.4 nm and 2–30 nm in size, or silver nitrate was added to cell culture media in the absence or presence of A549 cells as a model for human type II alveolar epithelial cells. The free silver ion activity measured after the addition of silver nanoparticles was determined by the initial ionic silver content. The p {Ag⁺} values indicated that the cell culture media decrease the free silver ion activity due to binding of silver ions by constituents of the media. In the presence of A549 cells, the free silver ion activity was further reduced. The morphology of A549 cells, cultivated in DME medium containing 9.1% (v/v) FBS, was affected by adding AgNO₃ at concentrations of ≥30 μM after 24 h. In comparison, silver nanoparticles up to a concentration of 200 μM Ag did not affect cellular morphology. Our experiments indicate that the effect of silver nanoparticles is mainly mediated by silver ions. An effect of silver on cellular morphology was observed at p {Ag⁺} ≤ 9.2.     

Toxicity of amorphous silica nanoparticles in mouse keratinocytes

The present study was designed to examine the uptake, localization, and the cytotoxic effects of well-dispersed amorphous silica nanoparticles in mouse keratinocytes (HEL-30). Mouse keratinocytes were exposed for 24 h to various concentrations of amorphous silica nanoparticles in homogeneous suspensions of average size distribution (30, 48, 118, and 535 nm SiO₂) and then assessed for uptake and biochemical changes. Results of transmission electron microscopy revealed all sizes of silica were taken up into the cells and localized into the cytoplasm. The lactate dehydrogenase (LDH) assay shows LDH leakage was dose- and size-dependent with exposure to 30 and 48 nm nanoparticles. However, no LDH leakage was observed for either 118 or 535 nm nanoparticles. The mitochondrial viability assay (MTT) showed significant toxicity for 30 and 48 nm at high concentrations (100 μg/mL) compared to the 118 and 535 nm particles. Further studies were carried out to investigate if cellular reduced GSH and mitochondria membrane potential are involved in the mechanism of SiO₂ toxicity. The redox potential of cells (GSH) was reduced significantly at concentrations of 50, 100, and 200 μg/mL at 30 nm nanoparticle exposures. However, silica nanoparticles larger than 30 nm showed no changes in GSH levels. Reactive oxygen species (ROS) formation did not show any significant change between controls and the exposed cells. In summary, amorphous silica nanoparticles below 100 nm induced cytotoxicity suggest size of the particles is critical to produce biological effects.     

Rapid extra-/intracellular biosynthesis of gold nanoparticles by the fungus <Emphasis Type="Italic">Penicillium</Emphasis> sp.

In this work, the fungus Penicillium was used for rapid extra-/intracellular biosynthesis of gold nanoparticles. AuCl₄ ⁻ ions reacted with the cell filtrate of Penicillium sp. resulting in extracellular biosynthesis of gold nanoparticles within 1 min. Intracellular biosynthesis of gold nanoparticles was obtained by incubating AuCl₄ ⁻ solution with fungal biomass for 8 h. The gold nanoparticles were characterized by means of visual observation, UV–Vis absorption spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). The extracellular nanoparticles exhibited maximum absorbance at 545 nm in UV–Vis spectroscopy. The XRD spectrum showed Bragg reflections corresponding to the gold nanocrystals. TEM exhibited the formed spherical gold nanoparticles in the size range from 30 to 50 nm with an average size of 45 nm. SEM and TEM revealed that the intracellular gold nanoparticles were well dispersed on the cell wall and within the cell, and they are mostly spherical in shape with an average diameter of 50 nm. The presence of gold was confirmed by EDX analysis.     

Structuring of Surfaces with Gold Nanoparticles by Using Bessel‐Like Beams

Here, the structuring of surfaces with gold nanoparticles by using Bessel‐like beam array is demonstrated. The experimental results show that the fabricated microring structures containing gold nanoparticles have a surface plasmon resonance in the spectral range of 520–540 nm, which can be tuned by selecting the laser treatment parameters. Fabricated microring structures exhibit a lower light transmittance comparing with the randomly distributed gold nanoparticles for wavelengths 500–570 nm due to the growth in the size of nanoparticles. In the spectral range of 600–700 nm, the light transmittance through microring structures is higher compared with the randomly distributed gold nanoparticles because of the removal of gold nanoparticles as gold has high reflectivity for wavelengths longer than 600 nm. The demonstrated method enables an easy fabrication of microring structures having tunable plasmonic properties.     

Synthetic and biogenic magnetite nanoparticles for tracking of stem cells and dendritic cells

Accurate delivery of cells to target organs is critical for success of cell-based therapies with stem cells or immune cells such as antigen-presenting dendritic cells (DC). Labeling with contrast agents before implantation provides a powerful means for monitoring cellular migration using magnetic resonance imaging (MRI). In this study, we investigated the uptake of fully synthesized or bacterial magnetic nanoparticles (MNPs) into hematopoietic Flt3⁺ stem cells and DC from mouse bone marrow. We show that (i) uptake of both synthetic and biogenic nanoparticles into cells endow magnetic activity and (ii) low numbers of MNP-loaded cells are readily detected by MRI.     

CW laser-induced photothermal conversion and shape transformation of gold nanodogbones in hydrated chitosan films

We investigate the photothermal conversion and transformation of gold nanoparticles with an initial dogbone shape after dispersion in hydrated chitosan films, which is a representative model of biological tissue, and excitation by a CW diode laser for 1 min. Gold nanodogbones are observed to undergo a distinct modification above a sharp threshold of ~11 W cm⁻² and 110 °C. Surprisingly, the very same modification is achieved up to at least 36 W cm⁻² and 250 °C. We use an analytical model derived from Gans theory to associate the change in color of the films with the change in shape statistics of these gold nanoparticles. This model proves both convenient and dependable. We interpret the photothermal transformation as a rearrangement of particles with a dogbone shape and an aspect ratio of 4.1 into rods with an aspect ratio of 2.5, where material from the end lobes of the dogbones may relocate to the waists of the rods. In turn, additional transitions to stable gold nanospheres may exhibit fairly slower kinetics.     

Shape evolution of gold nanoparticles

The tetraoctylammonium bromide-stabilized gold nanoparticles have been successfully fabricated. The shape evolution of these nanoparticles under different annealing temperatures has been investigated using high-resolution transmission electron microscopy. After an annealing at 100 °C for 30 min, the average diameters of the gold nanoparticles change a little. However, the shapes of gold nanoparticles change drastically, and facets appear in most nanoparticles. After an annealing at 200 °C for 30 min, not only the size but also the shape changes a lot. After an annealing at 300 °C for 30 min, two or more gold nanoparticles coalesce into bigger ones. In addition, because of the presence of Cu grid during the annealing, some gold particles become the nucleation sites of Cu₂O nanocubes, which possess a microstructure of gold-particle core/Cu₂O shell. These Au/Cu₂O heterostructure nanocubes can only be formed at a relatively high temperature (≥300 °C). The results can provide some insights on controlling the shapes of gold nanoparticles.