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33 result(s) for "Minaev, Nikita"
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Cell spheroid fusion: beyond liquid drops model
Biological self-assembly is crucial in the processes of development, tissue regeneration, and maturation of bioprinted tissue-engineered constructions. The cell aggregates—spheroids—have become widely used model objects in the study of this phenomenon. Existing approaches describe the fusion of cell aggregates by analogy with the coalescence of liquid droplets and ignore the complex structural properties of spheroids. Here, we analyzed the fusion process in connection with structure and mechanical properties of the spheroids from human somatic cells of different phenotypes: mesenchymal stem cells from the limbal eye stroma and epithelial cells from retinal pigment epithelium. A nanoindentation protocol was applied for the mechanical measurements. We found a discrepancy with the liquid drop fusion model: the fusion was faster for spheroids from epithelial cells with lower apparent surface tension than for mesenchymal spheroids with higher surface tension. This discrepancy might be caused by biophysical processes such as extracellular matrix remodeling in the case of mesenchymal spheroids and different modes of cell migration. The obtained results will contribute to the development of more realistic models for spheroid fusion that would further provide a helpful tool for constructing cell aggregates with required properties both for fundamental studies and tissue reparation.
Theoretical and Experimental Assay of Shock Experienced by Yeast Cells during Laser Bioprinting
Laser-induced forward transfer (LIFT) is a useful technique for bioprinting using gel-embedded cells. However, little is known about the stresses experienced by cells during LIFT. This paper theoretically and experimentally explores the levels of laser pulse irradiation and pulsed heating experienced by yeast cells during LIFT. It has been found that only 5% of the cells in the gel layer adjacent to the absorbing Ti film should be significantly heated for fractions of microseconds, which was confirmed by the fact that a corresponding population of cells died during LIFT. This was accompanied by the near-complete dimming of intracellular green fluorescent protein, also observed in response to heat shock. It is shown that microorganisms in the gel layer experience laser irradiation with an energy density of ~0.1–6 J/cm2. This level of irradiation had no effect on yeast on its own. We conclude that in a wide range of laser fluences, bioprinting kills only a minority of the cell population. Importantly, we detected a previously unobserved change in membrane permeability in viable cells. Our data provide a wider perspective on the effects of LIFT-based bioprinting on living organisms and might provide new uses for the procedure based on its effects on cell permeability.
THz generation from laser-induced breakdown in pressurized molecular gases: on the way to terahertz remote sensing of the atmospheres of Mars and Venus
The present paper studies the generation of terahertz (THz) radiation in CO2 in comparison with atmospheric air at a wide range of pressures. We established experimentally and explained theoretically that for these gases there are optimal pressures at about 1 bar for air and 0.5 bar for CO2 under which the efficiency of conversion from near-infrared to THz frequencies is the highest. We consider the possibility of applying femtosecond laser-induced THz generation for the study of the atmosphere of Mars and found that the overall THz yield near the surface of Mars is just a factor of 6 lower than on Earth. Comparable THz energy on the two planets is associated with underdense plasma on Earth (∼10% of neutrals) and full double ionization of carbon dioxide on Mars (∼200% of neutrals), the latter opening great perspective for THz remote sensing of trace gases in the Martian atmosphere.
Polysaccharide-based aerogels fabricated via supercritical fluid drying: a systematic review
Due to their biocompatibility and biodegradability, polysaccharide-based aerogels produced through supercritical fluid drying attract a lot of attention in the food industry, pharmaceuticals, tissue regeneration, etc. The aim of this study is to systematically review the types of polysaccharides used for aerogel fabrication, the approaches to their gelation before supercritical fluid drying, and the conditions of the drying. The literature screening was conducted according to the PRISMA protocol to give a full picture. Aerogels can be made using a wide range of polysaccharides (alginates, chitosan, agar, cellulose, carrageenans, starch, pectin, chitin, b -glucans, xanthan, guar gum, gellan gum, hyaluronic acid, tragacanth gum, locust bean gum), as well as polysaccharide-based biomaterials and artificial composites containing inorganic substances. The natural origin of the polysaccharides and a diversity of their chemical structure allow to explore a wide range of processing conditions for fabrication of aerogels for various applications. Different strategies to dissolve and gel the polysaccharides, aiming to reduce a number of required steps to aerogel formation and control its morphology, were reported. An analysis of the protocols used for supercritical drying of the formed gels showed trends toward process simplification and the implementation of one-pot strategies, as well as the better control over aerogel morphology, which open up opportunities for a wide application of polysaccharide-based aerogels in practice.
Endovenous laser coagulation: asymmetrical heat transfer and coagulation (modeling in blood plasma)
The objective of this study was to describe the dynamics of blood plasma heating and coagulation processes carried out by continuous laser radiation with wavelengths 1.55 and 1.94 μm through bare-tip fibers and fibers with radial output (radial fibers) used for endovenous laser coagulation (EVLC). The study was performed in previously thawed frozen donor blood plasma using high-speed shooting of the heating process through the shadow optical method. It has been shown that in the case of highly water-absorbed laser radiations, convection, explosive, and small-bubble boiling play a major role in the process of heat transfer and coagulation. It has been shown that in the case of radiation with wavelength λ = 1.94 μm, effective heat transfer begins at significantly lower levels of power compared to radiations with λ = 1.55 μm. It has been established that heat transfer is sharply asymmetrical and is directed mainly upwards and forwards (bare-tip fiber) or upwards (radial fibers). For a wavelength of 1.94 μm, the effect of self-cleaning of the fiber surface from coagulated plasma fragments was found. Except for short-term acts of explosive boiling, the heat transfer is asymmetrical and directed mainly upwards. This effect should lead to uneven heating and thermal damage to the vein wall with the maximum at its upper part. For EVLC, the use of radiation with a wavelength of 1.94 μm is more efficient and safer.
Endovenous laser coagulation: asymmetrical heat transfer (modeling in water)
The objective of this study was to describe the dynamics of water heating carried out by continuous laser radiation with wavelengths 1.47, 1.55, and 1.94 μm with different types of fibers used for endovenous laser coagulation. The study was conducted in water using high-speed surveying of the heating process through the shadow optical method. It has been shown that in the case of highly water-absorbed laser radiations, convection and boiling play a major role in the process of heat transfer. It has been shown that in the case of radiation with λ = 1.94 μm that is heavily absorbed by water, effective heat transfer begins at significantly lower levels of power compared to the weaker-absorbed radiations with λ = 1.47 and 1.55 μm. Mathematical models based only on thermal conductivity inadequately describe the process of real heat transfer during endovenous laser coagulation. It has been established that heat transfer is sharply asymmetrical and is directed mainly up-and-forward (bare-tip fiber) or upward (“radial” and “two-ring” fibers). Heat transfer for laser light with wavelength 1.94 μm is most effective than for 1.47 and 1.55 μm.
Optical Diagnostics of Supercritical CO2 and CO2-Ethanol Mixture in the Widom Delta
The supercritical CO2 (scCO2) is widely used as solvent and transport media in different technologies. The technological aspects of scCO2 fluid applications strongly depend on spatial–temporal fluctuations of its thermodynamic parameters. The region of these parameters’ maximal fluctuations on the p-T (pressure-temperature) diagram is called Widom delta. It has significant practical and fundamental interest. We offer an approach that combines optical measurements and molecular dynamics simulation in a wide range of pressures and temperatures. We studied the microstructure of supercritical CO2 fluid and its binary mixture with ethanol in a wide range of temperatures and pressures using molecular dynamics (MD) simulation. MD is used to retrieve a set of optical characteristics such as Raman spectra, refractive indexes and molecular refraction and was verified by appropriate experimental measurements. We demonstrated that in the Widom delta the monotonic dependence of the optical properties on the CO2 density is violated. It is caused by the rapid increase of density fluctuations and medium-sized (20–30 molecules) cluster formation. We identified the correlation between cluster parameters and optical properties of the media; in particular, it is established that the clusters in the Widom delta acts as a seed for clustering in molecular jets. MD demonstrates that the cluster formation is stronger in the supercritical CO2-ethanol mixture, where the extended binary clusters are formed; that is, the nonlinear refractive index significantly increased. The influence of the supercritical state in the cell on the formation of supersonic cluster jets is studied using the Mie scattering technique.
Laser Bioprinting with Cell Spheroids: Accurate and Gentle
Laser printing with cell spheroids can become a promising approach in tissue engineering and regenerative medicine. However, the use of standard laser bioprinters for this purpose is not optimal as they are optimized for transferring smaller objects, such as cells and microorganisms. The use of standard laser systems and protocols for the transfer of cell spheroids leads either to their destruction or to a significant deterioration in the quality of bioprinting. The possibilities of cell spheroids printing by laser-induced forward transfer in a gentle mode, which ensures good cell survival ~80% without damage and burns, were demonstrated. The proposed method showed a high spatial resolution of laser printing of cell spheroid geometric structures at the level of 62 ± 33 µm, which is significantly less than the size of the cell spheroid itself. The experiments were performed on a laboratory laser bioprinter with a sterile zone, which was supplemented with a new optical part based on the Pi-Shaper element, which allows for forming laser spots with different non-Gaussian intensity distributions. It is shown that laser spots with an intensity distribution profile of the “Two rings” type (close to Π-shaped) and a size comparable to a spheroid are optimal. To select the operating parameters of laser exposure, spheroid phantoms made of a photocurable resin and spheroids made from human umbilical cord mesenchymal stromal cells were used.
Controlled Structure of Polyester/Hydroxyapatite Microparticles Fabricated via Pickering Emulsion Approach
Biodegradable polyester/hydroxyapatite microparticles are widely proposed as microcarriers for drug/cell delivery or scaffolds for bone tissue regeneration. The current research implements the surfactant-free approach for the fabrication of polyester-based microparticles filled with hydroxyapatite nanoparticles (nHA) via the oil/water Pickering emulsion solvent evaporation technique for the first time, to the best of our knowledge. The process of polyester microparticle fabrication using nHA for the oil/water interface stabilization was studied as a function of phase used for nHA addition, which allows the preparation of a range of microparticles either filled with nHA or having it as a shell over the polymeric core. The effect of processing conditions (polymer nature, polymer/nHA ratio, ultrasound treatment) on particles’ total yield, size distribution, surface and volume morphology, and chemical structure was analyzed using SEM, EDX, Raman spectroscopy, and mapping. Addition of nHA either within the aqueous or oil phase allowed the effective stabilization of the oil/water interface without additional molecular surfactants, giving rise to hybrid microparticles in which total yield, size distribution, and surface morphology depended on all studied processing conditions. Preliminary ultrasound treatment of any phase before the emulsification process led to a complex effect but did not affect the homogeneity of nHA distribution within the polymeric core of the hybrid microparticles.
Evolution of Shock-Induced Pressure in Laser Bioprinting
Laser bioprinting with gel microdroplets that contain living cells is a promising method for use in microbiology, biotechnology, and medicine. Laser engineering of microbial systems (LEMS) technology by laser-induced forward transfer (LIFT) is highly effective in isolating difficult-to-cultivate and uncultured microorganisms, which are essential for modern bioscience. In LEMS the transfer of a microdroplet of a gel substrate containing living cell occurs due to the rapid heating under the tight focusing of a nanosecond infrared laser pulse onto thin metal film with the substrate layer. During laser transfer, living organisms are affected by temperature and pressure jumps, high dynamic loads, and several others. The study of these factors’ role is important both for improving laser printing technology itself and from a purely theoretical point of view in relation to understanding the mechanisms of LEMS action. This article presents the results of an experimental study of bubbles, gel jets, and shock waves arising in liquid media during nanosecond laser heating of a Ti film obtained using time-resolving shadow microscopy. Estimates of the pressure jumps experienced by microorganisms in the process of laser transfer are performed: in the operating range of laser energies for bioprinting LEMS technology, pressure jumps near the absorbing film of the donor plate is about 30 MPa. The efficiency of laser pulse energy conversion to mechanical post-effects is about 10%. The estimates obtained are of great importance for microbiology, biotechnology, and medicine, particularly for improving the technologies related to laser bioprinting and the laser engineering of microbial systems.