Raman spectroscopy to assess the differentiation of bone marrow mesenchymal stem cells into a glial phenotype.

Background: Mesenchymal stem cells (MSCs) are multipotent precursor cells with the ability to self-renew and differentiate into multiple cell linage, including the Schwann-like fate that promotes regeneration after lesion. Raman spectroscopy provides a precise characterization of the osteogenic, adipogenic, hepatogenic and myogenic differentiation of MSCs. However, the differentiation of bone marrow mesenchymal stem cells (BMSCs) towards a glial phenotype (Schwann-like cells) has not been characterized before using Raman spectroscopy. Method: We evaluated three conditions: 1) cell culture from rat bone marrow undifferentiated (uBMSCs), and two conditions of differentiation; 2) cells exposed to olfactory ensheathing cells-conditioned medium (dBMSCs) and 3) cells obtained from olfactory bulb (OECs). uBMSCs phenotyping was confirmed by morphology, immunocytochemistry and flow cytometry using antibodies of cell surface: CD90 and CD73. Glial phenotype of dBMSCs and OECs were verified by morphology and immunocytochemistry using markers of Schwann-like cells and OECs such as GFAP, p75 NTR and O4. Then, the Principal Component Analysis (PCA) of Raman spectroscopy was performed to discriminate components from the high wavenumber region between undifferentiated and glial-differentiated cells. Raman bands at the fingerprint region also were used to analyze the differentiation between conditions. Results: Differences between Raman spectra from uBMSC and glial phenotype groups were noted at multiple Raman shift values. A significant decrease in the concentration of all major cellular components, including nucleic acids, proteins, and lipids were found in the glial phenotype groups. PCA analysis confirmed that the highest spectral variations between groups came from the high wavenumber region observed in undifferentiated cells and contributed with the discrimination between glial phenotype groups. Conclusion: These findings support the use of Raman spectroscopy for the characterization of uBMSCs and its differentiation in the glial phenotype.

Characterization and Polydispersity of Volcanic Ash Nanoparticles in Synthetic Lung Fluid.

The inhalation of natural nanoparticles (NPs) emitted from volcanic activity may be a risk to human health. However, the literature rarely reports the fate and response of NPs once in contact with lung fluids. In this work, we studied the particle size distribution of ashfall from Popocatépetl volcano, Mexico. The collected ashes (n = 5) were analyzed with scanning electron microscopy (SEM) to obtain the elemental composition and morphology, and to determine the size of the ash particles using ParticleMetric software (PMS). The PMS reported most of the ash to have submicrometric size (<1 µm) and an average equivalent circle of 2.72 µm. Moreover, to our knowledge, this study investigated for the first time the behavior of ash NPs at different times (0 to 24 h) while in contact with in vitro lung fluid, Gamble Solution (GS) and Artificial Lysosomal Fluid (ALF) using dynamic light scattering (DLS). We found a large variability in the hydrodynamic diameter, with values less than 1 nm and greater than 5 µm. Furthermore, aggregation and disaggregation processes were recognized in GS and ALF, respectively. The results of this study increase the knowledge of the interaction between NPs and lung fluids, particularly within the alveolar macrophage region.

Nanoscale Changes on RBC Membrane Induced by Storage and Ionizing Radiation: A Mini-Review.

The storage lesions and the irradiation of blood cellular components for medical procedures in blood banks are events that may induce nanochanges in the membrane of red blood cells (RBCs). Alterations, such as the formation of pores and vesicles, reduce flexibility and compromise the overall erythrocyte integrity. This review discusses the alterations on erythrocytic lipid membrane bilayer through their characterization by confocal scanning microscopy, Raman, scanning electron microscopy, and atomic force microscopy techniques. The interrelated experimental results may address and shed light on the correlation of biomechanical and biochemical transformations induced in the membrane and cytoskeleton of stored and gamma-irradiated RBC. To highlight the main advantages of combining these experimental techniques simultaneously or sequentially, we discuss how those outcomes observed at micro- and nanoscale cell levels are useful as biomarkers of cell aging and storage damage.

Identification of refractory zirconia from catalytic converters in dust: An emerging pollutant in urban environments.

Using catalytic converters is one of the most effective methods to control vehicle emissions. A washcoat of cerium oxide-zirconia (CeO2-ZrO2) has been used to enhance the performance of the catalytic converter device. To date, the prevalence of this material in the environment has not been assessed. In this study, we present evidence of the existence of inhalable zirconia in urban dust. Samples of the washcoat, exhaust pipe, topsoil, and road dust were analyzed by X-ray fluorescence, X-ray diffraction (XRD), scanning electron microscopy (SEM), Raman spectroscopy, photoluminescence (PL) spectroscopy, and thermally stimulated luminescence (TSL). The results showed a CeO2-ZrO2 phase separation after sintering. This causes the emission of ZrO2, CeO2, and CeZrOx particles smaller than 1 µm, which can likely reach the alveolar macrophages in the lungs. The Ce-Zr content in road dust exceeds geogenic levels, and a significant correlation of 0.87 (p < 0.05) reflects a common anthropic source. Chronic exposure to such refractory particles may result in the development of non-occupational respiratory diseases. The inhalable crystalline compounds emitted by vehicles are a significant environmental health hazard, revealing the need for further investigation and assessment of zirconia levels generated by automobiles in urban areas worldwide.

Metal bioaccessibility, particle size distribution and polydispersity of playground dust in synthetic lysosomal fluids.

Inhalation of playground dust-derived fine particles in schoolyards poses a risk from exposure to metal(oids) and minerals. In this work, we obtained the total concentration and bioaccessibility of metal(oids) with Gamble Solution (GS) and Artificial Lysosomal Fluid (ALF) synthetic solutions, simulating the extracellular neutral pH environment of the lung and the intracellular conditions of the macrophage, respectively. Scanning Electron Microscope (SEM), and Dynamic Light Scattering analysis (DLS) techniques were used to characterize particles with a size smaller than 2.5 µm, which can be assimilated by macrophages in the deep part of the lung. Arsenic (As), lead (Pb), copper (Cu), manganese (Mn), zinc (Zn), and iron (Fe) showed concentrations of 39.9, 147.9, 286, 1369, 2313, 112,457 mg·kg-1, respectively. The results indicated that all studied elements were enriched when compared to (i) local geochemical background and (ii) findings reported in other cities around the world. Bioaccessibility of metal(oids) in GS was low-moderate for most studied elements. However, in ALF assays, bioaccessibility was high among the samples: for lead (Pb = 34-100%), arsenic (As = 14.7-100%), copper (Cu = 17.9-100%), and zinc (Zn = 35-52%) possibly related to hydrophobic minerals in dust. SEM and DLS image analysis showed that playground dust particles smaller than 2.5 µm are dominant, particularly particles with a size range of 500-600 nm. The polydispersity detected in these particle sizes showed that most of them might be crystalline compounds (elongated shapes) forming agglomerates instead of combustion particles (spheres). Moreover, the circularity detected varies from 0.57 to 0.79 (low roundness), which corroborates this finding. The presence of agglomerates of ultrafine/nanoparticles containing highly bioaccessible metals in playground sites may have severe implications in children's health. Therefore, further studies are required to characterize the size distribution, structure, shape and composition of such minerals which are essential factors related to the toxicology of inhaled dust particles.

Identification of inhalable rutile and polycyclic aromatic hydrocarbons (PAHs) nanoparticles in the atmospheric dust.

Addressing the presence of rutile nanoparticles (NPs) in the air is a work in progress, and the development of methodologies for the identification of NPs in atmospheric dust is essential for the assessment of its toxicological effects. To address this issue, we selected the fast growing desertic city of Hermosillo in northern Mexico. Road dust (n = 266) and soils (n = 10) were sampled and bulk Ti-contents were tested by portable X-ray fluorescence. NPs were extracted from atmospheric dust by PM1.0-PTFE filters and further characterized by Confocal Raman Microscopy, Energy-dispersive X-ray spectroscopy (EDS) coupled to Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). Results showed (i) the average concentration of Ti in road dust (3447 mg kg-1) was similar to natural values and worldwide urban dusts; (ii) the bulk geochemistry was not satisfactory for Ti-NPs identification; (iii) 76% of the total extracted PM1.0 sample corresponded to NPs; (iv) mono-microaggregates of rutile NPs were identified; (v) ubiquitous polycyclic aromatic hydrocarbons (PAHs) were linked to NPs. The genotoxicity of rutile and PAHs, in connection with NPs content, make us aware of a crucial emerging environmental issue of significant health concern, justifying further research in this field.

Subtissue thermal sensing based on neodymium-doped LaF3 nanoparticles.

In this work, we report the multifunctional character of neodymium-doped LaF3 core/shell nanoparticles. Because of the spectral overlap of the neodymium emission bands with the transparency windows of human tissues, these nanoparticles emerge as relevant subtissue optical probes. For neodymium contents optimizing the luminescence brightness of Nd³âº:LaF3 nanoparticles, subtissue penetration depths of several millimeters have been demonstrated. At the same time, it has been found that the infrared emission bands of Nd³âº:LaF3 nanoparticles show a remarkable thermal sensitivity, so that they can be advantageously used as luminescent nanothermometers for subtissue thermal sensing. This possibility has been demonstrated in this work: Nd³âº:LaF3 nanoparticles have been used to provide optical control over subtissue temperature in a single-beam plasmonic-mediated heating experiment. In this experiment, gold nanorods are used as nanoheaters while thermal reading is performed by the Nd³âº:LaF3 nanoparticles. The possibility of a real single-beam-controlled subtissue hyperthermia process is, therefore, pointed out.