The invention of a scanning electron microscopy (SEM) pushed the imaging methods and allowed for the observation of cell details with a high resolution. Currently, SEM appears as an extremely useful tool to analyse the morphology of biological samples. The aim of this paper is to provide a set of guidelines for using SEM to analyse morphology of prokaryotic and eukaryotic cells, taking as model cases Escherichia coli bacteria and B-35 rat neuroblastoma cells. Herein, we discuss the necessity of a careful sample preparation and provide an optimised protocol that allows to observe the details of cell ultrastructure (≥ 50 nm) with a minimum processing effort. Highlighting the versatility of morphometric descriptors, we present the most informative parameters and couple them with molecular processes. In this way, we indicate the wide range of information that can be collected through SEM imaging of biological materials that makes SEM a convenient screening method to detect cell pathology.
The aspiration to interact living cells with electronics challenges researchers to develop materials working at the interface of these two distinct environments. A successful interfacing coating should exhibit both biocompatibility and desired functionality of a bio-integrated device. Taking into account biodiversity, the tissue interface should be fine-tuned to the specific requirements of the bioelectronic systems. In this study, we pointed to electrochemical doping of conducting polymers as a strategy enabling the efficient manufacturing of interfacing platforms, in which features could be easily adjusted. Consequently, we fabricated conducting films based on a poly(3,4-ethylenedioxythiophene) (PEDOT) matrix, with properties modulated through doping with selected ions: PSS− (poly(styrene sulfonate)), ClO4− (perchlorate), and PF6− (hexafluorophosphate). Striving to extend the knowledge on the relationships governing the dopant effect on PEDOT films, the samples were characterized in terms of their chemical, morphological, and electrochemical properties. To investigate the impact of the materials on attachment and growth of cells, rat neuroblastoma B35 cells were cultured on their surface and analyzed using scanning electron microscopy and biological assays. Eventually, it was shown that through the choice of a dopant and doping conditions, PEDOT-based materials can be efficiently tuned with diversified physicochemical properties. Therefore, our results proved electrochemical doping of PEDOT as a valuable strategy facilitating the development of promising tissue interfacing materials with characteristics tailored as required.
Two highly efficient commercial organic photosensitizers—azure A (AA) and 5-(4-aminophenyl)-10,15,20-(triphenyl)porphyrin (APTPP)—were covalently attached to the glass surface to form a photoactive monolayer. The proposed straightforward strategy consists of three steps, i.e., the initial chemical grafting of 3-aminopropyltriethoxysilane (APTES) followed by two chemical postmodification steps. The chemical structure of the resulting mixed monolayer (MIX_TC_APTES@glass) was widely characterized by X-ray photoelectron (XPS) and Raman spectroscopies, while its photoactive properties were investigated in situ by UV–Vis spectroscopy with α-terpinene as a chemical trap. It was shown that both photosensitizers retain their activity toward light-activated generation of reactive oxygen species (ROS) after immobilization on the glassy surface and that the resulting nanolayer shows high stability. Thanks to the complementarity of the spectral properties of AA and APTPP, the effectiveness of the ROS photogeneration under broadband illumination can be optimized. The reported light-activated nanocoating demonstrated promising antimicrobial activity toward Escherichia coli (E. coli), by reducing the number of adhered bacteria compared to the unmodified glass surface.
Due to its biocompatibility and advantageous electrochemical properties, platinum is commonly used in the design of biomedical devices, e.g., surgical instruments, as well as electro-medical or orthopedic implants. This article verifies the hypothesis that a thin layer of sputter-coated platinum may possess antibacterial effects. The purpose of this research was to investigate the adhesion and growth ability of a model strain of Gram-negative bacteria, Escherichia coli, on a surface of a platinum-coated glass slide. Although some previous literature reports suggests that a thin layer of platinum would inhibit the formation of bacterial biofilm, the results of this study suggest otherwise. The decrease in the number of bacterial cells attached to the platinum-coated glass, which was observed within first three hours of culturing, was found to be a short-time effect, vanishing after 24 h. Consequently, it was shown that a thin layer of sputter-coated platinum did not exhibit any antibacterial effect. For this reason, this study indicates an urgent need for the development of new methods of surface modification that could reduce bacterial surface colonization of platinum-based biomedical devices.
Vascular regeneration is a complex process, additionally limited by the low regeneration potential of blood vessels. Hence, current research is focused on the design of artificial materials that combine biocompatibility with a certain rate of biodegradability and mechanical robustness. In this paper, we have introduced a scaffold material made of poly(L-lactide-co-glycolide)/poly(isosorbide sebacate) (PLGA/PISEB) fibers fabricated in the course of an electrospinning process, and confirmed its biocompatibility towards human umbilical vein endothelial cells (HUVEC). The resulting material was characterized by a bimodal distribution of fiber diameters, with the median of 1.25 µm and 4.75 µm. Genotyping of HUVEC cells collected after 48 h of incubations on the surface of PLGA/PISEB scaffolds showed a potentially pro-angiogenic expression profile, as well as anti-inflammatory effects of this material. Over the course of a 12-week-long hydrolytic degradation process, PLGA/PISEB fibers were found to swell and disintegrate, resulting in the formation of highly developed structures resembling seaweeds. It is expected that the change in the scaffold structure should have a positive effect on blood vessel regeneration, by allowing cells to penetrate the scaffold and grow within a 3D structure of PLGA/PISEB, as well as stabilizing newly-formed endothelium during hydrolytic expansion.