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Osteoarthritis (OA) is a degenerative joint disease with limited therapeutic options, where inflammation plays a critical role in disease progression. Extracellular vesicles (EV) derived from mesenchymal stromal cells (MSC) have shown potential as a therapeutic approach for OA by modulating inflammation and alleviating degenerative processes in the joint. This study evaluated the therapeutic effects for the treatment of OA of two types of EV-exosomes and matrix-bound nanovesicles (MBV)-both derived from the human umbilical cord MSC (UC-MSC) via differential ultracentrifugation. Different phenotypes of human monocyte-derived macrophages (MDM) were used to study the anti-inflammatory properties of EV in vitro, and the medial meniscectomy-induced rat model of knee osteoarthritis (MMx) was used in vivo. The study found that both EV reduced pro-inflammatory cytokines IL-6 and TNF-α in MDM. However, exosomes showed superior results, preserving the extracellular matrix (ECM) of hyaline cartilage, and reducing synovitis more effectively than MBVs. Additionally, exosomes downregulated inflammatory markers (TNF-α, iNOS) and increased Arg-1 expression in macrophages and synovial fibroblasts, indicating a stronger anti-inflammatory effect. These results suggest UC-MSC exosomes as a promising therapeutic option for OA, with the potential for modulating inflammation and promoting joint tissue regeneration.

Recent years have witnessed significant advancements in the cryopreservation of various tissues and cells, yet several challenges persist. This review evaluates the current state of cryopreservation, focusing on contemporary methods, notable achievements, and ongoing difficulties. Techniques such as slow freezing and vitrification have enabled the successful preservation of diverse biological materials, including embryos and ovarian tissue, marking substantial progress in reproductive medicine and regenerative therapies. These achievements highlight improved post-thaw survival and functionality of cryopreserved samples. However, there are remaining challenges such as ice crystal formation, which can lead to cell damage, and the cryopreservation of larger, more complex tissues and organs. This review also explores the role of cryoprotectants and the importance of optimizing both cooling and warming rates to enhance preservation outcomes. Future research priorities include developing new cryoprotective agents, elucidating the mechanisms of cryoinjury, and refining protocols for preserving complex tissues and organs. This comprehensive overview underscores the transformative potential of cryopreservation in biomedicine, while emphasizing the necessity for ongoing innovation to address existing challenges., Competing Interests: The authors declare no conflicts of interest.

Despite a number of advantages of ion-selective optical sensors (optodes), their practical application is limited by their response mechanism, which leads to the dependence of the signal on the activity of two ions (analyte ion and the so-called reference ion) in the solution at the same time. Here, we show that the introduction of a lipophilic electrolyte into the polymeric optode membrane allows assessing the ionic activity of H + cations regardless of the concentration of the background electrolyte containing a hydrophilic cation, with NaCl as an example of such an electrolyte. For the first time, the applicability of this approach is proven theoretically utilizing the numerical simulation of optode response. A correlation between the interfacial potential stability and the single-ion optical response is established. The predicted optical response is independent of background cation concentration to a significant extent. Theoretical conclusions are supported by experimental data obtained with chromoionophore-based optodes doped with various lipophilic electrolytes, including ionic liquids, by thin-film spectrophotometry and macrophotography coupled with digital color analysis. Most of the experimental sensor characteristics, such as the response range and its median, as well as its independence from the background electrolyte concentration are in quantitative agreement with the proposed theoretical description.

To achieve a transition from conventional liquid-junction reference electrodes (LJF REs) to their all-solid-state alternatives, organic electrolytes are often introduced into the polymeric electrode membranes. In this article, we implement a theoretical approach to the explanation and quantification of the boundary potential stabilization phenomenon for the electrodes modified with organic electrolytes (Q + B - ). For the first time, stabilization of the phase boundary potential due to the partition of lipophilic ions of the Q + B - electrolyte between the polymeric and aqueous phases is numerically simulated to predict the LJF electrodes behavior. The impact of the hydrophilic electrolyte on the potential stabilization is demonstrated both numerically and experimentally. The developed model predicted that the small additions of a traditional ion-exchanger enhance performance of the Q + B - -based reference electrodes. For some specific cases, the optimal concentrations of Q + B - and ion-exchanger in the polymeric phase are suggested to provide stable electrode potential in a broad range of aqueous electrolyte concentrations. In addition, the efficiency of the stabilization was shown to be dependent on the overall Q + B - load in the polymeric membrane rather than on the closeness of the partition coefficients of the Q + and B - ions; and on the volume of the aqueous phase. The model results are verified experimentally with poly(vinyl chloride) membranes containing ion-exchanger or hydrophilic electrolyte and Q + B - in various proportions. A good agreement between the measured electrode response and the theoretical results is observed in a broad range of solution concentrations. In particular, the cationic function of membranes containing KTpClPB is suppressed, and the electrodes begin to behave as REs starting from 50-60 mol. % of ETH500 electrolyte added to the membrane, relative to the total amount of salt.

Biodegradable polymeric fibrous non-woven materials are widely used type of scaffolds for tissue engineering. Their morphology and properties could be controlled by composition and fabrication technology. This work is aimed at development of fibrous scaffolds from a multicomponent polymeric system containing biodegradable synthetic (polylactide, polycaprolactone) and natural (gelatin, chitosan) components using different methods of non-woven mats fabrication: electrospinning and electro-assisted solution blow spinning. The effect of the fabrication technique of the fibrous materials onto their morphology and properties, including the ability to support adhesion and growth of cells, was evaluated. The mats fabricated using electrospinning technology consist of randomly oriented monofilament fibers, while application of solution blow spinning gave a rise to chaotically arranged multifilament fibers. Cytocompatibility of all fabricated fibrous mats was confirmed using in vitro analysis of metabolic activity, proliferative capacity and morphology of NIH 3T3 cell line. Live/Dead assay revealed the formation of the highest number of cell-cell contacts in the case of multifilament sample formed by electro-assisted solution blow spinning technology.

Here, we propose a novel approach to the classification of blue ballpoint pen inks based on a combination of selective extraction of coloring components from a paper carrier, digital color analysis (DCA) of the remaining traces, and hierarchical cluster analysis of DCA results. Since most documents of high importance are still produced in hard copies, the proposed method, being highly time- and cost-efficient, could be a significant contribution to forensic science in the field of authenticating handwritten documents. Several commonly used solvents were applied in parallel as extractants to the replicate strokes produced by each pen. It turned out to be possible to limit the number of extractants required for an unambiguous classification to three. We have shown that the optimal descriptor for agglomerative clustering is the colorimetric distance between the original and extracted ink traces in the RGB color space. Five separate clusters of inks that are independent of sample storage temperature were obtained from a set of 16 different pens. This conclusion was further confirmed by the analysis of principal components. The developed DCA-based data processing pipeline outperformed the clustering based on the data of high-performance liquid chromatography in terms of versatility providing a more informative analysis with respect to the inks based on the phthalocyanine dyes.

Ionic liquids (ILs) have a wide variety of applications in modern electrochemistry due to their unique electrolytic properties. In particular, they are promising candidates as dopants for polymeric membranes in potentiometric sensors and liquid-junction free reference electrodes. However, the effective use of ILs requires a comprehensive understanding of their electrolytic behavior in the polymeric phase. We report here the exploration of the electrolytic and diffusion properties of IL 1-hexyl-3-methyl-1H-imidazol-3-ium bis[(trifluoromethyl)sulfonyl]amide ([C 6 Meim][NTf 2 ]) in a poly(vinyl chloride) matrix. Chronopotentiometry is utilized to determine the concentration of charge carriers, ionic diffusion coefficients and apparent dissociation constant of [C 6 Meim][NTf 2 ] in PVC membranes plasticized with a mixture of [C 6 Meim][NTf 2 ] and bis(2-ethylhexyl) sebacate (DOS) over a wide range of IL concentrations. The diffusion properties of [C 6 Meim][NTf 2 ] are confirmed by NMR-diffusometry. The non-monotonic electrolytic behavior of the IL in PVC-DOS matrix is described for the first time. A maximum ionization degree and diffusion coefficient is observed at 30 wt.% of IL in the plasticizing mixture. Thus, it is shown that by varying the flexible parameter of the IL to plasticizer ratio in the polymeric phase one can tune the electrolytic and transport properties of sensing PVC membranes.

We report here a simple and cost-effective technique for classification of the samples of writing inks based on their digital color analysis (DCA). The dynamics of artificial aging of writing inks under UV irradiation was studied by means of DCA for the first time. The color of ballpoint pen marks was recorded over time using an ordinary consumer DSLR camera. The inks were classified according to the parameters of their color degradation curves with precision superior to conventional Raman scattering method, which serves as a proof-of-concept of the proposed approach. The reported approach has broad prospects for implementation by criminalists for document investigation when document forgery is suspected. The proposed technique can be of interest not only in the field of forensic science but also for those who deal with dyes and dye-containing materials and their degradation over time as well as for the study of any processes, the evolution of which is reflected in a color change.

Here, we report on systematic investigation of the impact of coextraction of the aqueous electrolyte and anion interference on the response of cation-selective bulk optodes. It is evident that to deliberately manage the properties of chemical sensors and to apply them in routine analysis, one should have exhaustive insight into their operation mechanism. Despite the extensive research in the field of ionophore-based optodes and numerous attempts for their practical application, the understanding of how coextraction of an aqueous electrolyte influences its response characteristics has not been developed thus far. Meanwhile, the electrolyte coextraction determines the detection limits of analogous ion-selective electrodes. A theoretical model based on phase distribution equilibrium is proposed to quantitatively describe the effect of Donnan exclusion failure on the response of polymeric plasticized optodes. The theoretical conclusions are confirmed by the results obtained with Na + /pH-selective optodes based on a neutral chromoionophore as a model system in solutions containing anions of various lipophilicities (Cl - , NO 3 - , I - , SCN - ). For the first time, it is shown that coextraction leads to a significant shift of the response range of the optodes as well as to nonmonotonic response curves due to the transition from cationic to anionic response. An approach to estimate the coextraction constants of electrolytes from the optode response curves is proposed. The limitations in the applicability of optodes due to co-ion interference are explored. It is found that neglecting anion interference can cause dramatic errors in the results of analyses with optical sensors.4 - ). For the first time, it is shown that coextraction leads to a significant shift of the response range of the optodes as well as to nonmonotonic response curves due to the transition from cationic to anionic response. An approach to estimate the coextraction constants of electrolytes from the optode response curves is proposed. The limitations in the applicability of optodes due to co-ion interference are explored. It is found that neglecting anion interference can cause dramatic errors in the results of analyses with optical sensors.

The indicator-based polymeric color standards for color referencing in digital color analysis (DCA) of optical chemical sensors (optodes) are proposed. In the novel standards, the colors referring to the actual absorption bands of the protonated and deprotonated forms of the indicator are mixed in constant proportions. The standards are based on the lipophilic pH-indicators: ETH5350 and ETH2439, commonly used in optodes, and the lipophilic electrolyte TBATBB. The dependence of the standard color on the TBATBB concentration in the optode is established and found to be linear. The standard colors remain unchanged upon varying the solution pH and the nature and the concentration of the electrolyte. Calibration curves of the indicator pH-optodes obtained in horse serum and referenced to the developed standards demonstrate lower error to span ratio, broader span and higher sensitivity as compared to the same data processed with the conventional gray standard. The colorimetric signal of the pH-optodes array measured in serum sample and referenced to the developed standards allowed accurate determination of the sample pH thus demonstrating practical prospects of the proposed color standards., (Copyright © 2019. Published by Elsevier B.V.)

The concept of reducing the analysis time with the bulk sensors operating in nonequilibrium mode is formulated theoretically and verified experimentally with ion-selective polymeric optodes. The influence of the measurement time on the optical response range, span, median, and sensitivity is calculated and demonstrated experimentally. Model aqueous samples are successfully analyzed using a nonequilibrium measurement protocol with conventional sodium-selective optodes and with pH optodes containing lipophilic organic electrolyte. The analysis time is significantly reduced from 300 to ca. 100 s, with sufficient recovery, reproducibility, and precision of the sample determination.

Linear Nernstian response is obtained for a neutral ionophore-based Ca(2+)-selective electrode down to 10(-10) M CaCl2 by means of galvanostatic polarization. The densities of the applied cathodic current were tuned for particular concentrations of Ca2+. The procedure included recording the potential at zero current, followed by measurements when current is passed through the electrode, and then again at zero current. The respective chronopotentiometric curves included negative ohmic drop immediately after turning the current on, the polarization domain, and positive ohmic drop when the current was turned off, followed with the relaxation domain. The potentials immediately after the positive ohmic drops were used as analytical signals. These potentials make a straight line with Nernstian slope when currents are tuned (optimized) for each particular concentration. An iteration procedure is proposed which allows for simultaneous optimization of the current density and accessing analyte concentration in the sample.