Centre for Nanomaterials and Biotechnology

Extracellular vesicles (EVs) are emerging as critical mediators of intercellular communication in the tumor microenvironment (TME), profoundly influencing cancer progression. These nano-sized vesicles, released by both tumor and stromal cells, carry a diverse cargo of proteins, nucleic acids, and lipids, reflecting the dynamic cellular landscape and mediating intricate interactions between cells. This review provides a comprehensive overview of the biogenesis, composition, and functional roles of EVs in cancer, highlighting their significance in both basic research and clinical applications. We discuss how cancer cells manipulate EV biogenesis pathways to produce vesicles enriched with pro-tumorigenic molecules, explore the specific contributions of EVs to key hallmarks of cancer, such as angiogenesis, metastasis, and immune evasion, emphasizing their role in shaping TME and driving therapeutic resistance. Concurrently, we submit recent knowledge on how the cargo of EVs can serve as a valuable source of biomarkers for minimally invasive liquid biopsies, and its therapeutic potential, particularly as targeted drug delivery vehicles and immunomodulatory agents, showcasing their promise for enhancing the efficacy and safety of cancer treatments. By deciphering the intricate messages carried by EVs, we can gain a deeper understanding of cancer biology and develop more effective strategies for early detection, targeted therapy, and immunotherapy, paving the way for a new era of personalized and precise cancer medicine with the potential to significantly improve patient outcomes.

Plant extracellular vesicles (pEVs) derived from numerous edible sources gain a lot of attention in recent years, mainly due to the potential to efficiently carry bioactive molecules into mammalian cells. In the present study, we focus on isolation of PDNVs (plant-derived nanovesicles) and pEVs from callus culture and from BY-2 culture of Nicotiana tabacum (tobacco). Tobacco was selected as a source of plant vesicles, as it is commonly used by human, moreover it is a model organism with established techniques for cultivation of explant cultures in vitro. Explant cultures are suitable for the isolation of pEVs in large quantities, due to their fast growth in sterile conditions. As the efficiency of isolation methods varies, we were comparing two methods of isolation. We evaluated biophysical and biochemical properties of plant vesicles, as well as differences between isolates. We encountered difficulties in the form of vesicles aggregation, which is often described in publications focused on mammalian nanovesicles. In an effort to prevent vesicle aggregation, we used trehalose in different stages of isolation. We show tobacco-derived vesicles successfully enter tobacco and mesenchymal cell lines. We observed that tobacco-nanovesicles isolated by different methods incorporated fluorescent dye with different efficiency. The results of our study show tobacco-derived vesicles isolated by various isolation methods are able to enter plant, as well as mammalian cells.

The urgent need for sustainable and innovative approaches to mitigate the increasing levels of atmospheric CO2 necessitates the development of efficient methods for its removal. In this study, we focus on the new, innovative approach for synthesis and functionalization of metal-organic framework (MOF) ZIF-8 in one step at room temperature to enhance its capacity for CO2 capture. Specifically, we investigated the impact of four amino-compounds, namely tetraethylenepentamine (TEPA), hexadecylamine (HDA), ethanolamine (ELA), and cyclopropylamine (CPA), on the chemical structure, size, surface area and porosity, and CO2 capturing of ZIF-8 powder. By varying concentrations of the amino-compounds, we examined their influence on the ZIF-8 properties. Our findings demonstrate that each amino-compound and its respective concentration exhibit distinct effects on the characteristics of ZIF-8. Notably, the ZIF-8 sample functionalized with the highest presented concentration of TEPA exhibited significant improvement in CO2 trapping efficiency, with a 33.3% enhancement. Moreover, least concentrated samples with added HDA or CPA demonstrated notable improvements with enhancements of 46.6% and 18.6%, respectively. These results highlight the potential of simple synthesis and functionalization techniques for MOFs in enhancing their CO2 capture capabilities. The findings from this study offer new opportunities for the development of strategies to mitigate CO2 emissions using MOFs.

Cortical projection neurons polarize and form an axon while migrating radially. Even though these dynamic processes are closely interwoven, they are regulated separately-the neurons terminate their migration when reaching their destination, the cortical plate, but continue to grow their axons. Here, we show that in rodents, the centrosome distinguishes these processes. Newly developed molecular tools modulating centrosomal microtubule nucleation combined with in vivo imaging uncovered that dysregulation of centrosomal microtubule nucleation abrogated radial migration without affecting axon formation. Tightly regulated centrosomal microtubule nucleation was required for periodic formation of the cytoplasmic dilation at the leading process, which is essential for radial migration. The microtubule nucleating factor γ-tubulin decreased at neuronal centrosomes during the migratory phase. As distinct microtubule networks drive neuronal polarization and radial migration, this provides insight into how neuronal migratory defects occur without largely affecting axonal tracts in human developmental cortical dysgeneses, caused by mutations in γ-tubulin.

Stability and cytotoxicity of PEGylated Au NPs is crucial for biomedical application. In this study, we have focused on thermal stability of PEGylated Au NPs at 4 and 37 °C and after sterilization in autoclave. Gold nanoparticles were prepared by direct sputtering of gold into PEG and PEG-NH 2 . Transmission electron microscopy revealed that NPs exhibit a spherical shape with average dimensions 3.8 nm for both AuNP_PEG and AuNP_PEG-NH 2 . The single LSPR band at wavelength of 509 nm also confirmed presence of spherical Au NPs in both cases. Moreover, according to UV–Vis spectra, the Au NPs were overall stable during aging or thermal stressing and even after sterilization in autoclave. Based on gel electrophoresis results, the higher density of functionalizing ligands and the higher stability is assumed on AuNP_PEG-NH 2 . Changes in concentration of gold did not occur after thermal stress or with aging. pH values have to be adjusted to be suitable for bioapplications – original pH values are either too alkaline (AuNP_PEG-NH 2 , pH 10) or too acidic (AuNP_PEG, pH 5). Cytotoxicity was tested on human osteoblasts and fibroblasts. Overall, both Au NPs have shown good cytocompatibility either freshly prepared or even after Au NPs′ sterilization in the autoclave. Prepared Au NP dispersions were also examined for their antiviral activity, however no significant effect was observed. We have synthesized highly stable, non-cytotoxic PEGylated Au NPs, which are ready for preclinical testing.

The solvent casting method was used for five types of polyvinylidene difluoride (PVDF) nanocomposite film preparation. The effect of nanofillers in PVDF nanocomposite films on the structural, phase, and friction and mechanical properties was examined and compared with that of the natural PVDF film. The surface topography of PVDF nanocomposite films was investigated using a scanning electron microscope (SEM) and correlative imaging (CPEM, combinate AFM and SEM). A selection of 2D CPEM images was used for a detailed study of the spherulitic morphologies (grains size around 6–10 μm) and surface roughness (value of 50–68 nm). The chemical interactions were evaluated by Fourier transform infrared spectroscopy (FTIR). Dominant polar γ-phase in the original PVDF, PVDF_ZnO and PVDF_ZnO/V, the most stable non-polar α-phase in the PVDF_V_CH nanocomposite film and mixture of γ and α phases in the PVDF_V and PVDF_ZnO/V_CH nanocomposite films were confirmed. Moderately hydrophilic PVDF nanocomposite films with water contact angle values (WCA) in the range of 58°–69° showed surface stability with respect to the Zeta potential values. The effect of positive or negative Zeta-potential values of nanofillers (ζn) on the resulting negative Zeta-potential values (ζ) of PVDF nanocomposite films was demonstrated. Interaction of PVDF chains with hydroxy groups of vermiculite and amino and imino groups of CH caused transformation of γ-phase to α. The friction properties were evaluated based on the wear testing and mechanical properties were evaluated from the tensile tests based on Young’s modulus (E) and tensile strength (Rm) values. Used nanofillers caused decreasing of friction and mechanical properties of PVDF nanocomposite material films.

Abstract In the evolving landscape of cancer research, 3D cell cultures, particularly tumor cell spheroids, are increasingly preferred in drug screening due to their enhanced mimicry of in vivo tumor environments, especially in drug resistance aspects. However, the consistent formation of uniform spheroids and their precise manipulation remain complex challenges. Among various methodologies, droplet microfluidics emerges as a highly effective approach for tumor spheroid formation. This paper introduces a novel, multifaceted microfluidic system that streamlines the entire spheroid cultivation process: (i) generating tumor spheroids from cell suspensions within individual droplets, (ii) merging these droplets into a continuous aqueous phase once spheroid formation is complete, and (iii) transferring the spheroids to a specialized cultivation area within the chip, equipped with trapping elements for extended cultivation in perfusion mode. Remarkably, this process requires no hydrogel encapsulation or external handling, as all operations are conducted within the microfluidic chip. Fabricated from the innovative OSTE+ (off-stoichiometry thiol-ene epoxy) polymer, the chip is designed for repeated use. To show its efficacy, we successfully formed spheroids from MCF-7, GAMG, and U87 cell lines in our system and compared them with spheroids prepared by a traditional agarose microwell method. Additionally, our methodology has successfully enabled the in-chip release of spheroids from droplets, followed by their effective trapping for subsequent cultivation, a process we have exemplified with MCF-7 spheroids. To our knowledge, this research represents the first instance of a fully integrated droplet microfluidic platform achieving scaffoldless tumor spheroid formation and handling. Our method holds promise for improving high-throughput, automated procedures in the formation, transfer, and cultivation of tumor cell spheroids. Graphical abstract

High Resolution Image Download MS PowerPoint Slide Transition metal (TM) sulfides belong to the class of 2D materials with a wide application range. Various methods, including solvothermal, hydrothermal, chemical vapor deposition, and quartz ampoule-based approaches, have been employed for the synthesis of TM sulfides. Some of them face limitations due to the low stability of TM sulfides and their susceptibility to oxidation, and others require more sophisticated equipment or complex and rare precursors or are not scalable. In this work, we propose an alternative approach for the synthesis of 2D TM sulfides by sulfurization of corresponding metal oxides in the vapor of CS 2 at elevated temperature. Subsequent treatment in liquid nitrogen allows exfoliation of created sulfides to a 2D structure. A proposed approach was successfully applied to nine transition metals: Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, and W. The resulting materials were extensively characterized using various analytical techniques with a focus on their crystalline structure and 2D nature. Our approach offers several advantages including the use of simple precursors (CS 2 and metal oxides), universality (in all cases, the sulfides were obtained), equipment simplicity (tube furnace and quartz reactor), short preparation time (3 h), and the ability of morphology and phase tuning (in particular cases) of the created materials by adjusting the temperature. In addition, gram-scale bulk materials can be obtained in the entry-level laboratories using the proposed approach.

High Resolution Image Download MS PowerPoint Slide The properties of MXene flakes, a new class of two-dimensional materials, are strictly determined by their surface termination. The most common termination groups are oxygen-containing (═O or –OH) and fluorine (−F), and their relative ratio is closely related to flake stability and catalytic activity. The surface termination can vary significantly among MXene flakes depending on the preparation route and is commonly determined after flake preparation by using X-ray photoelectron spectroscopy (XPS). In this paper, as an alternative approach, we propose the combination of surface-enhanced Raman spectroscopy (SERS) and artificial neural networks (ANN) for the precise and reliable determination of MXene flakes’ (Ti 3 C 2 T x ) surface chemistry. Ti 3 C 2 T x flakes were independently prepared by three scientific groups and subsequently measured using three different Raman spectrometers, employing resonant excitation wavelengths. Manual analysis of the SERS spectra did not enable accurate determination of the flake surface termination. However, the combined SERS-ANN approach allowed us to determine the surface termination with a high accuracy. The reliability of the method was verified by using a series of independently prepared samples. We also paid special attention to how the results of the SERS-ANN method are affected by the flake stability and differences in the conditions of flake preparation and Raman measurements. This way, we have developed a universal technique that is independent of the above-mentioned parameters, providing the results with accuracy similar to XPS, but enhanced in terms of analysis time and simplicity.

The Off-Stoichiometry Thiol-ene and Epoxy (OSTE+) polymer technology has been increasingly utilised in the field of microfluidics and lab-on-a-chip applications. However, the impact of OSTEMER polymers, specifically the OSTEMER 322 formulation, on cell viability has remained limited. In this work, we thoroughly explored the biocompatibility of this commercial OSTEMER formulation, along with various surface modifications, through a broad range of cell types, from fibroblasts to epithelial cells. We employed cell viability and confluence assays to evaluate the performance of the material and its modified variants in cell culturing. The properties of the pristine and modified OSTEMER were also investigated using surface characterization methods including contact angle, zeta potential, and X-ray photoelectron spectroscopy. Mass spectrometry analysis confirmed the absence of leaching constituents from OSTEMER, indicating its safety for cell-based applications. Our findings demonstrated that cell viability on OSTEMER surfaces is sufficient for typical cell culture experiments, suggesting OSTEMER 322 is a suitable material for a variety of cell-based assays in microfluidic devices.

Neurons rely on the microtubule cytoskeleton to create and maintain their sophisticated cellular architectures. Advances in cryogenic electron microscopy, expansion microscopy, live imaging, and gene editing have enabled novel insights into mechanisms of centrosomal and acentrosomal microtubule nucleation, the key process generating new microtubules. This has paved the way for the functional dissection of distinct microtubule networks that regulate various processes during neuronal development, including neuronal delamination, polarization, migration, maturation, and synapse function. We review recent progress in understanding the molecular concepts of microtubule nucleation, how these concepts underlie neurodevelopmental processes, and pinpoint the open questions. Since microtubules play a pivotal role in axon regeneration within the adult central nervous system, understanding the processes of microtubule nucleation could inform strategies to enhance the regenerative capabilities of neurons in the future.

in the absence of plasmon triggering). A set of control experiments and quantum chemical modeling revealed that the plasmon assistance proceeds through hot electron excitation followed by an interaction with an enamine with the formation of anion radical species. We also demonstrated the high stability of the proposed system in multiple catalytic cycles without leaching metal ions, which makes our approach especially promising for heterogeneous asymmetric photocatalysis.

Extracellular vesicles (EVs) are lipid-enclosed structures that facilitate intercellular communication by transferring cargo between cells. Although predominantly studied in mammals, extracellular vesicles are ubiquitous across metazoans, and thus research in non-mammalian models is critical for fully elucidating extracellular vesicles biology. Recent advances demonstrate that extracellular vesicles mediate diverse physiological processes in non-mammalian vertebrates, including fish, amphibians, and reptiles. Piscine extracellular vesicles promote fin regeneration in zebrafish and carry heat shock proteins regulated by stress. Frog extracellular vesicles containing microRNAs modulate angiogenesis, while turtle extracellular vesicles coordinate reproductive functions. Venom from snakes contains extracellular vesicles that mirror the whole venom composition and interact with mammalian cells. Invertebrates also possess extracellular vesicles involved in immunity, development, and pathogenesis. Molluscan extracellular vesicles participate in shell formation and host interactions. Arthropod models, including Drosophila, genetically dissect conserved pathways controlling extracellular vesicles biogenesis and signalling. Nematode extracellular vesicles regulate larval development, animal communication, and ageing via conserved extracellular vesicles proteins. Ancient metazoan lineages utilise extracellular vesicles as well, with cnidarian extracellular vesicles regulating immunity and regeneration. Ultimately, expanding extracellular vesicles research beyond typical biomedical models to encompass phylogenetic diversity provides an unparalleled perspective on the conserved versus specialised aspects of metazoan extracellular vesicles roles over ∼500 million years. With a primary focus on the literature from the past 5 years, this review aims to reveal fundamental insights into EV-mediated intercellular communication mechanisms shaping animal physiology.

Amorphous and crystalline nickel‐based metal‐organic frameworks (Ni‐MOFs) were prepared via a one‐pot synthesis at room temperature in methanol using 2‐methylimidazole as a ligand. The crystallinity was adjusted by varying the solvent volume, yielding an amorphous phase with higher surface area (≈242 m 2 g −1 ) and a crystalline form with reduced porosity (≈22 m 2 g −1 ). Comprehensive structural, morphological, and spectroscopic analyses confirmed distinct coordination environments, particle sizes and colloidal behaviors. Gas sorption measurements revealed enhanced CO 2 uptake in the amorphous Ni‐MOF (≈9.5 cm 3 g −1 ) compared to the crystalline sample (≈3.4 cm 3 g −1 ), consistent with its greater pore volume and surface area. Photocatalytic degradation of methyl orange under 365 nm UV irradiation demonstrated faster activity for the amorphous material, with a pseudo‐first‐order rate constant of 0.0157 min −1 versus 0.0035 min −1 for the crystalline sample. These findings suggest that structural features such as higher surface area, pore volume, and possible disorder contribute to the improved gas sorption and photocatalytic response. The use of mild reaction conditions and a single solvent system offers a straightforward and energy‐efficient approach for preparing functional MOFs with tunable crystallinity, applicable in environmental remediation contexts.

Abstract Microfluidic devices play a crucial role in advanced cell biology applications, including cell separations, cultivations, migration and interaction studies, diagnostic devices, and organ‐on‐chips. One of the frequent purposes of such devices is the ability to selectively address the attachment of cells at defined locations on the surface. This study explores the application of middle‐energy carbon, oxygen, and nitrogen ions to locally modify the surface of cyclic olefin copolymer (COC) thermoplastic material, allowing selective cell growth on patterned polymer surfaces. The investigation considers ion element type, ion beam energy, and ion irradiation fluence, analyzing their influence on the modification effect. Characterization of the modified surfaces involves various surface‐analytical methods such as contact angle, energy dispersive spectroscopy (SEM‐EDX), atomic force microscopy (AFM), x‐ray photoelectron spectroscopy (XPS), rutherford backscattering spectrometry (RBS), and elastic recoil detection analysis (ERDA). The study extends to practical aspects, with a representative cancer cell line, MCF‐7, grown on the patterned surface to evaluate the degree of selective attachment. Additionally, the stability of the irradiated patterns is tested under elevated temperatures beyond the glass transition temperature ( T g ), demonstrating the compatibility of the approach with hot embossing technology. The findings underscore the potential of ion beam treatment for COC in cell‐biology‐related applications, offering insights into surface modification techniques for enhanced functionality in microfluidic devices.

The consideration of human and environmental exposure to dendrimers, including cytotoxicity, acute toxicity, and cell and tissue accumulation, is essential due to their significant potential for various biomedical applications. This study aimed to evaluate the biodistribution and toxicity of a novel methoxyphenyl phosphonium carbosilane dendrimer, a potential mitochondria-targeting vector for cancer therapeutics, in 2D and 3D cancer cell cultures and zebrafish embryos. We assessed its cytotoxicity (via MTT, ATP, and Spheroid growth inhibition assays) and cellular biodistribution. The dendrimer cytotoxicity was higher in cancer cells, likely due to its specific targeting to the mitochondrial compartment. In vivo studies using zebrafish demonstrated dendrimer distribution within the vascular and gastrointestinal systems, indicating a biodistribution profile that may be beneficial for systemic therapeutic delivery strategies. The methoxyphenyl phosphonium carbosilane dendrimer shows promise for applications in cancer cell delivery, but additional studies are required to confirm these findings using alternative labelling methods and more physiologically relevant models. Our results contribute to the growing body of evidence supporting the potential of carbosilane dendrimers as vectors for cancer therapeutics.

Manufacturing of membranes for carbon dioxide (CO 2 ) capture is a significant research topic. Achieving maximum CO 2 sorption capacity while maintaining air permeability with a minimum number of technological steps was the main motivation of this work. The greatest advantages of this approach are its simplicity, low cost and easy transition to industrial scale. Electrospun nanofibrous membranes polyacrylonitrile (PAN)/triethylenetetramine (TETA) and polyacrylonitrile (PAN)/tetraethylenepentamine (TEPA) were prepared by one-step technology (modifying amines TETA, TEPA in different weight concentrations dissolved directly in spinning solution) using two different spinning conditions: needle spinning (electric field attached to a hollow needle through which a polymer solution is extruded under pressure) and wire spinning (electric field connected to a thin wire that is coated with a layer of polymer solution, and the spinning thus takes place from the free surface). Wire electrospinning turns out to be more suitable for a one-step technology with a modifying substance in the spinning solution. The best result as to the CO 2 sorption capacity has been obtained for wire spinning PAN_TEPA_2% 11.7 ± 1.3 cm 3 /g with air permeability 53 ± 5 L/m 2 /s, which gives a good chance for the design of a sandwich functional unit for practical use. In addition to studies of CO 2 adsorption, the article also deals with the comparison of both spinning methods for the PAN polymer, which have not yet been compared for this polymer in the literature, not only from the point of view of the possibility of preparing PAN nanofibers, but also their functional use precisely for CO 2 capture.

Abstract We report the approach of successful immobilization of iron oxide particles (IOPs) on UV‐activated surface of polymers. PA6 (polyamide 6), PTFE (polytetrafluoroethylene), UHMWPE (ultra‐high‐molecular‐weight polyethylene), and PP (polypropylene) foils were studied. The surface of polymers was firstly activated by UV radiation and subsequently grafted with IOPs. We have also investigated the effect of surface pre‐grafting with chitosan and triethanolamine on subsequent IOPs grafting. IOPs have been synthesized by co‐precipitation method of ferric and ferrous ions. The changes in surface properties of modified polymer foils before/after individual steps of modification were characterized by contact angle determination, electrokinetic analysis, X‐ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM). The contact angle decreased after UV activation due to the formation of oxygen polar groups. The presence of iron on the polymer surfaces, revealed by XPS, showed that IOPs were successfully bound to the polymer surfaces. XRD analysis confirmed that the prepared IOPs were in the form of magnetite (Fe3O4). The best results were obtained for the procedure containing UV surface activation, chitosan grafting, and subsequent IOPs immobilization.

This study explores novel membranes based on deep eutectic solvents (DES) combined with commercial polyvinyl chloride (PVC) for bioenergy generation in microbial fuel cells (MFCs) utilizing wastewater. The incorporation of DES into these systems provides a cost-effective and environmentally sustainable approach for fabricating exchange membranes, thanks to their unique properties, including ionic conductivity, solvation capabilities, ease of fabrication, and biodegradability. Polymer membranes were synthesized using a solution casting technique, incorporating a DES composed of Aliquat-336 (trioctylmethylammonium chloride) and menthol in a 1:1 M ratio. The final membrane compositions included the eutectic phase at percentages of 30 %, 50 %, and 70 %, which were compared to pristine PVC membranes. Characterization techniques such as scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), ionic exchange capacity (IEC), ion transport number, water uptake and swelling ratio were employed to assess membrane properties. This study focuses on MFCs used for wastewater treatment and energy recovery, highlighting the potential of DES-based membranes as a more sustainable alternative to traditional materials and establishing a new avenue for the use of DES in bioelectrochemical applications. Specifically, the membranes containing 50 % DES exhibited promising performance, demonstrating the potential of DES as an active phase in membrane technology, with a power density of 292.2 mW m −3 and a COD removal rate of 78.3 %.

Modern tissue engineering requires not only degradable materials promoting cell growth and differentiation, but also vascularization of the engineered tissue. Porous polylactide/polycaprolactone (PLA/PCL, ratio 3/5) foam scaffolds were prepared by a combined porogen leaching and freeze-drying technique using NaCl (crystal size 250–500 µm) and a water-soluble cellulose derivative (KlucelTM E; 10–100% w/w relative to the total PLA/PCL concentration) as porogens. Scanning electron microscopy, micro-CT, and Brunauer–Emmett–Teller analysis showed that all scaffolds contained a trimodal range of pore sizes, i.e., macropores (average diameter 298–539 μm), micropores (100 nm to 10 μm), and nanopores (mostly around 3.0 nm). All scaffolds had an open porosity of about 90%, and the pores were interconnected. The size of the macropores and the nanoporosity were higher in the scaffolds prepared with Klucel. Nanoporosity increased water uptake by the scaffolds, while macroporosity promoted cell ingrowth, which was most evident in scaffolds prepared with 25% Klucel. Human adipose-derived stem cells co-cultured with endothelial cells formed pre-vascular structures in the scaffolds, which was further enhanced in a dynamic cell culture system. The scaffolds are promising for the engineering of pre-vascularized soft tissues (relatively pliable 10% Klucel scaffolds) and hard tissues (mechanically stronger 25% and 50% Klucel scaffolds).