Evaluation of biophysical alterations in the epithelial and endothelial layer of patients with Bullous Keratopathy.

The cornea is a fundamental ocular tissue for the sense of sight. Thanks to it, the refraction of two-thirds of light manages to participate in the visual process and protect against mechanical damage. Because it is transparent, avascular, and innervated, the cornea comprises five main layers: Epithelium, Bowman's layer, stroma, Descemet's membrane, and endothelium. Each layer plays a key role in the functionality and maintenance of ocular tissue, providing unique ultrastructural and biomechanical properties. Bullous Keratopathy (BK) is an endothelial dysfunction that leads to corneal edema, loss of visual acuity, epithelial blisters, and severe pain, among other symptoms. The corneal layers are subject to changes in their biophysical properties promoted by Keratopathy. In this context, the Atomic Force Microscopy (AFM) technique in air was used to investigate the anterior epithelial surface and the posterior endothelial surface, healthy and with BK, using a triangular silicone tip with a nominal spring constant of 0.4 N/m. Six human corneas (n = 6) samples were used for each analyzed group. Roughness data, calculated by third-order polynomial adjustment, adhesion, and Young's modulus, were obtained to serve as a comparison and identification of morphological and biomechanical changes possibly associated with the pathology, such as craters and in the epithelial layer and exposure of a fibrotic layer due to loss of the endothelial cell wall. Endothelial cell membrane area and volume data were calculated, obtaining a relevant comparison between the control and patient. Such results may provide new data on the physical properties of the ocular tissue to understand the physiology of the cornea when it has pathology.

Physical, morphological and bioactive properties of Co-Cr-W-Ta alloys: Influence of insertion of tantalum and surface thermochemical treatment on bioactivity.

The development of bioactivity in bioinert metallic alloys is a field of interest aiming to improve some aspects of these materials for implant applications. New Co63 Cr28 W9-x Tax alloys with different Ta concentrations (x = 0, 2, 4, 6, and 9% w/w) were synthesized in the work reported here. The alloys were characterized by x-ray diffraction, volumetric density, Vickers microhardness, atomic force microscopy, scanning electron microscopy (SEM), and energy-dispersion x-ray spectroscopy (EDS). Bioactivity properties were evaluated by in vitro tests with simulated body fluid (SBF). In vivo assays were performed to assess biocompatibility. The influence of surface thermochemical treatment and Ta insertion on the bioactive properties of the alloys was investigated. The results showed that the alloy structure comprises εCo and αCo phases, with cobalt as a matrix with Cr, W, and Ta as a solid solution. TaCo2 phase is observed in the alloys with 4, 6, and 9% w/w of Ta, and its amount increase as Ta concentration increases. Volumetric density is reduced (from 8.78 ± 0.06 to 8.56 ± 0.09 g/cm3 ) as Ta concentration increases (from 0% to 9% w/w) mainly due to the lower density of the tantalum compared to the tungsten metal. On the other hand, the TaCo2 phase contributes to the increase of Vickers's hardness by ~17.6% for the alloy with 9% Ta (394.7 ± 8.1 HV) compared with Co63 Cr28 W9 (336 ± 5 HV). The topographic analysis showed increased roughness and adhesion due to the nucleation of Ta1.1 O1.05 and Ca2 Ta2 O7 crystals after surface thermochemical treatment. The roughness and adhesion increase from 16.9 ± 0.6 nm and 8.3 ± 1.8 nN (untreated surface) to 255.7 ± 17.7 nm and 24.1 ± 12.6 nN (treated surface), respectively, for the Co63 Cr28 Ta9 alloy. These results suggest that thermochemical treatment provides surface conditions favorable to hydroxyapatite (HA) nucleation. The SEM and EDS data showed the nucleation of spongy structures, consistent with HA, composed mainly of Ca and P, indicating that oxides tantalum promoted a bioactive response on the sample's surface. The biological assay corroborated the alloy's safety and applicability, highlighting its potential in biomedical application since no harmful effects were observed.

New Insights into Hemolytic Anemias: Ultrastructural and Nanomechanical Investigation of Red Blood Cells Showed Early Morphological Changes.

Several diseases are characterized by changes in the mechanical properties of erythrocytes. Hemolytic anemias are an example of these diseases. Among the hemolytic anemias, Sickle Cell Disease and Thalassemia are the most common, characterized by alterations in the structure of their hemoglobin. Sickle cell disease has a pathological origin in synthesizing abnormal hemoglobin, HbS. In contrast, thalassemia results in extinction or decreased synthesis of α and ß hemoglobin chains. This work presents a detailed study of biophysical and ultrastructural early erythrocytes membrane alterations at the nanoscale using Atomic Force Microscopy (AFM). Cells from individuals with sickle cell anemia and thalassemia mutations were studied. The analysis methodology in the AFM was given by blood smear and exposure of the inner membrane for ghost analysis. A robust statistic was used with 65,536 force curves for each map, ten cells of each type, with three individuals for each sample group. The results showed significant differences in cell rigidity, adhesion, volume, and roughness at early morphological alterations, bringing new perspectives for understanding pathogenesis. The sickle cell trait (HbAS) results stand out. Significant alterations were observed in the membrane properties, bringing new perspectives for the knowledge of this mutation. This work presents ultrastructural and biomechanical signatures of sickle cell anemia and thalassemia genotypes, which may help determine a more accurate biophysical description and clinical prognosis for these diseases.

SARS-CoV-2 Unrevealed: Ultrastructural and Nanomechanical Analysis.

The ongoing outbreak of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) started in late 2019 and spread across the world, infecting millions of people, with over 3.3 million deaths worldwide. To fight back the virus, it is necessary to understand how the main structures work, especially those responsible for the virus infectivity pathogenicity. Here, using the most advanced atomic force microscopy techniques, SARS-CoV-2 viral particles were analyzed, with a special focus on their ultrastructure, adsorption conformation, and nanomechanical behavior. The results uncovered the aspects of the organization and the spatial distribution of the proteins on the surface of the viral particles. It also showed the compliant behavior of the membrane and ability to recover from mechanical injuries. At least three layers composing the membrane and their thickness were measured, protecting the virus from external stress. This study provides new insight into the ultrastructure of SARS-CoV-2 particles at the nanoscale, offering new prospects that could be employed for mapping viral surfaces. The understanding of the viruses' capacity to survive mechanical disruptions at any level and their ability to recover from such injuries can shed a light on the structure-function relationship and help us to find targets for drug action, especially for this virus that, to this day, has no course of treatment approved.