Search Results (16)

Exosomes are extracellular nanovesicles secreted by cells that efficiently deliver therapeutic cargo for cancer treatment. However, because exosomes are present in low quantities and have limited target specificity, internal and external stress stimulation has been studied to increase exosome efficiency. Inspired by these studies, the uptake efficiency of cobalt chloride-induced hypoxic cancer cell-secreted exosomes was evaluated. Western blotting and RT-PCR data revealed increased exosome secretion and different protein compositions exhibited by hypoxic exosomes (H-Exos) compared to natural normoxic exosomes (N-Exos). Furthermore, these H-Exos were continuously stimulated using low-intensity ultrasound (LICUS) at an intensity of 360 mW/cm² and a frequency of 3 MHz in vitro and 1 MHz in vivo. Hyperthermic and mechanical stress caused by ultrasound successfully improved exosome uptake via clathrin-mediated pathways, and confocal laser microscopy showed strong internal localization near the target cell nuclei. Finally, LICUS-equipped H-Exos were loaded with hydrophobic curcumin (H-Exo-Cur) and used to treat parent HepG2 liver cancer cells. The UV–Vis spectrophotometer displayed enhanced stability, solubility, and concentration of the encapsulated drug molecules. In MTT and FACS studies, approximately 40 times higher cell death was induced, and in animal studies, approximately 10 times higher tumor sizes were suppressed by LICUS-assisted H-Exo-Cur compared to the control. In this study, the delivery platform constructed demonstrated enormous potential for liver cancer therapy. Full article

Nonmelanocytic skin cancers (NMSCs) are currently the most common group of human cancers and include all tumors that are not melanomas. Increased exposure to sunlight over the past few years, the lack of regular and proper use of sunscreen, the aging of the population, and better screening techniques are the reasons for the escalation in their diagnosis. Squamous cell carcinoma (SCC) comprises nearly 37% of the tumors in this group and can originate from actinic keratosis (AK), which usually presents as pink, often scaly plaques, usually located on the face or scalp. Advances in dermatoscopy, as well as the development of other non-invasive skin imaging modalities such as high-frequency ultrasound (HFUS), reflectance confocal microscopy (RCM), and optical coherence tomography (OCT), have allowed for greatly increased sensitivity in diagnosing these lesions and monitoring their treatment. Since AK therapy is usually local, and SCCs must be removed surgically, non-invasive imaging methods enable to correctly qualify difficult lesions. This is especially important given that they are very often located on the face, and achieving an appropriate cosmetic result after treatments in this area is very important for the patients. In this review, the authors describe the use of non-invasive skin imaging methods in the diagnosis of actinic keratosis. Full article

(1) Background: the aim of the study was to demonstrate its usefulness in the field of imaging evaluation of plaque morphology in psoriasis vulgaris, with an emphasis on the use of confocal microscopy and other advanced skin-imaging techniques. (2) Methods: we conducted a prospective study over two years (July 2022–April 2024), on patients diagnosed with moderate or severe psoriasis vulgaris, treated in the dermatology department of our institution. We selected 30 patients, of whom 15 became eligible according to the inclusion and the exclusion criteria. A total of 60 psoriasis plaques were analyzed by dermatoscopy using a Delta 30 dermatoscope and Vidix 4.0 videodermoscope (VD), by cutaneous ultrasound (US) using a high-resolution 20 MHz linear probe, and by confocal microscopy, along with histopathological analysis. (3) Results: the study included fifteen patients with vulgar psoriasis, diagnosed histopathologically, of whom six were women and nine were men, with an average age of 55. Between two and six plaques per patient were selected and a total of sixty psoriasis plaques were analyzed by non-invasive imaging techniques. Twelve lesions were analyzed with ex vivo fluorescence confocal microscopy (FCM), compared to histology. US showed that the hyperechoic band and the lack of damage to the subcutaneous tissue were the most common criteria. The epidermis and dermis were found to be thicker in the area of psoriasis plaques compared to healthy skin. Dermatoscopy showed that the specific aspect of psoriasis plaques localized on the limbs and trunk was a lesion with an erythematous background, with dotted vessels with regular distribution on the surface and covered by white scales with diffuse distribution. The presence of bushy vessels with medium condensation was the most frequently identified pattern on VD. Good correlations were identified between the histological criteria and those obtained through confocal microscopy. (4) Conclusions: the assessment and monitoring of patients with psoriasis vulgaris can be conducted in a more complete and all-encompassing manner by incorporating dermatoscopy, ultrasonography, and confocal microscopy in clinical practice. Full article

Individuals with diabetes at risk of developing diabetic kidney disease (DKD) are challenging to identify using currently available clinical methods. Prognostic accuracy and initiation of treatment could be improved by a quantification of the renal microvascular rarefaction and the increased vascular tortuosity during the development of DKD. Super-resolution ultrasound (SRUS) imaging is an in vivo technique capable of visualizing blood vessels at sizes below 75 µm. This preclinical study aimed to investigate the alterations in renal blood vessels’ density and tortuosity in a type 2 diabetes rat model, Zucker diabetic fatty (ZDF) rats, as a prediction of DKD. Lean age-matched Zucker rats were used as controls. A total of 36 rats were studied, subdivided into ages of 12, 22, and 40 weeks. Measured albuminuria indicated the early stage of DKD, and the SRUS was compared with the ex vivo micro-computed tomography (µCT) of the same kidneys. Assessed using the SRUS imaging, a significantly decreased cortical vascular density was detected in the ZDF rats from 22 weeks of age compared to the healthy controls, concomitant with a significantly increased albuminuria. Already by week 12, a trend towards a decreased cortical vascular density was found prior to the increased albuminuria. The quantified vascular density in µCT corresponded with the in vivo SRUS imaging, presenting a consistently lower vascular density in the ZDF rats. Regarding vessel tortuosity, an overall trend towards an increased tortuosity was present in the ZDF rats. SRUS shows promise for becoming an additional tool for monitoring and prognosing DKD. In the future, large-scale animal studies and human trials are needed for confirmation. Full article

The synthesis of new composite nanomaterials based on graphene oxide (GO)modified with cellulose and its derivatives, as well as nanocellulose, is currently an important direction and contributes toward solving many problems in various fields such as nanotechnology, information technology, medicine, high-dielectric materials, and nanoelectronics. In this work, for the first time, for the production of GO and its membrane with carboxymethylcellulose (CMC), local Kazakhstan “Ognevsky” graphite was used as the initial raw material. In this regard, the preparation of nanocomposites of GO modified with cellulose derivatives, including CMC, attracts great interest from scientists and expands its field of practical application due to the significant changes in its physicochemical properties. In this work, the GO obtained using the Hummers method was modified by CMC, and its physicochemical, structural, and electrical characteristics were studied. The GO/CMC membrane was synthesized by mixing 1% GO with crushed solid mass of CMC (0.03 g; 0.06 g; 0.15 g) and then processing using ultrasound. The surface morphology of the GO/CMC membrane was studied using scanning electron microscopy (SEM). It has been established that by increasing the mass of CMC (0.03 g; 0.06 g; 0.15 g), the polymerization of CMC occurs on the surface of GO nanosheets. Cross-sectional micrographs of GO/CMC show the formation of sandwich-like layered structures. The synthesis efficiency (yield) of GO from synthetic graphite is 10.8%, and GO from Ognevsky graphite is 11.9%, almost 1.1% more than GO from synthetic graphite. The mechanical tensile strength increases from 2.3 MPa to 14.3 MPa and the Young’s modulus from 2.3 MPa to 143 MPa. The electrical parameters of the humidity sensor based on GO and GO/CMC membranes (0.03 g; 0.06 g; 0.15 g) were studied as a function of humidity to determine the performance of the device. Full article

Imaging of the ocular vasculature can provide new insights into the pathophysiology of ocular diseases. This study proposes a novel high-frequency super-resolution ultrasound localization microscopy (SRULM) technique and evaluates its ability to measure in vivo perfusion changes in the rat eye at elevated intraocular pressure (IOP). A 38.4 MHz center frequency linear array transducer on a VisualSonics Vevo F2 imaging platform was used to collect high frame rate (1 kHz) radiofrequency data of the posterior rat eye following systemic microbubble contrast injection. Following clutter and spatiotemporal non-local means filtering, individual microbubbles were localized and tracked. The microbubble tracks were accumulated over 10,000 frames to generate vascular images quantifying perfusion velocity and direction. Experiments were performed using physiologic relevant controlled flow states for algorithm validation and subsequently performed in vivo on the rat eye at 10 mm Hg IOP increments from 10 to 60 mm Hg. The posterior vasculature of the rat eye, including the ophthalmic artery, long posterior ciliary arteries and their branches, central retinal artery and retinal arterioles and venules were successfully visualized, and velocities quantified at each IOP level. Significant reductions in arterial flow were measured as IOP was elevated. High-frequency SRULM can be used to visualize and quantify the perfusion velocity of the rat eye in both the retrobulbar and intraocular vasculature simultaneously. The ability to detect ocular perfusion changes throughout the depth of the eye may help elucidate the role ischemia has in the pathophysiology of ocular diseases such as glaucoma. Full article

In this paper, mechanical stirring and ultrasonic treatment are used to separate graphite electrode materials from copper foils in recycling spent lithium-ion batteries (LIBs). Firstly, the effects of ultrasonic power (60–180 W), ultrasonic time (1–8 min), stirring speed (420–2000 rpm), and stirring time (1–8 min) on the abscission rate of active material on copper foil were studied. It was found that the peeling-off ratio of electrode material under ultrasonic treatment was 91.34% compared with stirring treatment (84.22%). The removal of electrode material from copper foil during stirring was mainly through mechanical scrubbing. As a comparison, the generation of the microjets induced by ultrasound, the local high-temperature and high-pressure environment, and the free radicals during ultrasonic treatment are the key factors to further improve electrode material removal efficiency. An integrated ultrasound-mechanical stirrer technique can achieve a high-efficient separation performance (approximately 100% peeling-off ratio) of anode electrode materials from copper foils. The effects of mechanical stirring speed, temperature, and treatment time on the peeling-off ratios of the ultrasound-mechanical stirrer-assisted system were investigated. Finally, the results of XRF (X-ray fluorescence spectrometer), XRD (X-ray diffraction), and SEM-EDS (scanning electron microscopy coupled with energy dispersive X-ray spectroscopy) showed that the as-separated graphite electrode material had high purity and contained almost no copper foil impurities. Numerical simulation analyses briefly showed that the difference between pressure and ultrasonic temperature changes in the boundary between different anode layers (graphite on copper foil in aqueous solution) was the main effective factor in the considerable separation of graphite from copper anode foil under ultrasonic-assisted delamination. Full article

Ocular choroidal melanoma (OCM) is the most common ocular primary malignant tumor in adults, and there is an increasing emphasis on its early detection and treatment worldwide. The main obstacle in early detection of OCM is its overlapping clinical features with benign choroidal nevus. Thus, we propose ultrasound localization microscopy (ULM) based on the image deconvolution algorithm to assist the diagnosis of small OCM in early stages. Furthermore, we develop ultrasound (US) plane wave imaging based on three-frame difference algorithm to guide the placement of the probe on the field of view. A high-frequency Verasonics Vantage system and an L22-14v linear array transducer were used to perform experiments on both custom-made modules in vitro and a SD rat with ocular choroidal melanoma in vivo. The results demonstrate that our proposed deconvolution method implement more robust microbubble (MB) localization, reconstruction of microvasculature network in a finer grid and more precise flow velocity estimation. The excellent performance of US plane wave imaging was successfully validated on the flow phantom and in an in vivo OCM model. In the future, the super-resolution ULM, a critical complementary imaging modality, can provide doctors with conclusive suggestions for early diagnosis of OCM, which is significant for the treatment and prognosis of patients. Full article

In the quest for higher acquisition rates of ultrasound images, the simultaneous emission of encoded waves has the potential to overcome the trade-off between acquisition time and image quality. However, the lack of fully orthogonal codes has led to the use of forward models and inverse problem approaches to estimate the imaged medium. Nonetheless, due to some simplifying assumptions on which these models rely, the previously stated trade-off still appears in these acquisition/reconstruction schemes. In this paper, a forward model for ultrasound wave propagation inside a scattering medium is developed for the simultaneous coded emission of plane waves. The tissue reflectivity function of the imaged medium is estimated by solving an ${\ell }_1$-regularized version of the corresponding inverse problem. The proposed method is evaluated in silico and in vitro. We demonstrate that this method outperforms the conventional technique that consists of successive emissions of plane waves, reconstruction using delay and sum (DAS), and coherent compounding. In silico, the ability to separate close scatterers is improved by a factor of four in the axial direction and by a factor of 2.5 in the lateral direction. In vitro, the spatial resolution at −6 dB is decreased by a factor of seven. These results suggest that the proposed method could be a valuable tool, particularly for ultrasound imaging of sparse mediums such as in ultrasound localization microscopy. Full article

Obesity is a risk factor of chronic kidney disease (CKD), leading to alterations in the renal vascular structure. This study tested if renal vascular density and tortuosity was quantifiable in vivo in obese rats using microbubble-based super-resolution ultrasound imaging. The kidneys of two 11-week-old and two 20-week-old male obese Zucker rats were compared with age-matched male lean Zucker rats. The super-resolution ultrasound images were manually divided into inner medulla, outer medulla, and cortex, and each area was subdivided into arteries and veins. We quantified vascular density and tortuosity, number of detected microbubbles, and generated tracks. For comparison, we assessed glomerular filtration rate, albumin/creatinine ratio, and renal histology to evaluate CKD. The number of detected microbubbles and generated tracks varied between animals and significantly affected quantification of vessel density. In areas with a comparable number of tracks, density increased in the obese animals, concomitant with a decrease in glomerular filtration rate and an increase in albumin/creatinine ratio, but without any pathology in the histological staining. The results indicate that super-resolution ultrasound imaging can be used to quantify structural alterations in the renal vasculature. Techniques to generate more comparable number of microbubble tracks and confirmation of the findings in larger-scale studies are needed. Full article

Liquid metal convection plays an important role in natural and technical processes. In experimental studies, an instrumentation with a sub-millimeter spatial resolution is required in an opaque fluid to resolve the flow field near the boundary layer. Using ultrasound methods, the trade-off between the frequency and imaging depth of typical laboratory experiments limits the spatial resolution. Therefore, the method of ultrasound localization microscopy (ULM) was introduced in liquid metal experiments for the first time in this study. To isolate the intrinsic scattering particles, an adaptive nonlinear beamformer was applied. As a result, an average spatial resolution of 188 μm could be achieved, which corresponded to a fraction of the ultrasound wavelength of 0.28. A convection experiment was measured using ULM. Due to the increased spatial resolution, the high-velocity gradients and the recirculation areas of a liquid metal convection experiment could be observed for the first time. The presented technique paves the way for in-depth flow studies of convective turbulent liquid metal flows that are close to the boundary layer. Full article

Super-resolution ultrasound imaging, based on the localization and tracking of single intravascular microbubbles, makes it possible to map vessels below 100 µm. Microbubble velocities can be estimated as a surrogate for blood velocity, but their clinical potential is unclear. We investigated if a decrease in microbubble velocity in the arterial and venous beds of the renal cortex, outer medulla, and inner medulla was detectable after intravenous administration of the α1-adrenoceptor antagonist prazosin. The left kidneys of seven rats were scanned with super-resolution ultrasound for 10 min before, during, and after prazosin administration using a bk5000 ultrasound scanner and hockey-stick probe. The super-resolution images were manually segmented, separating cortex, outer medulla, and inner medulla. Microbubble tracks from arteries/arterioles were separated from vein/venule tracks using the arterial blood flow direction. The mean microbubble velocities from each scan were compared. This showed a significant prazosin-induced velocity decrease only in the cortical arteries/arterioles (from 1.59 ± 0.38 to 1.14 ± 0.31 to 1.18 ± 0.33 mm/s, p = 0.013) and outer medulla descending vasa recta (from 0.70 ± 0.05 to 0.66 ± 0.04 to 0.69 ± 0.06 mm/s, p = 0.026). Conclusively, super-resolution ultrasound imaging makes it possible to detect and differentiate microbubble velocity responses to prazosin simultaneously in the renal cortical and medullary vascular beds. Full article

Ceramic Matrix Composites (CMC) are promising materials for high-temperature applications where damage tolerant failure behavior is required. Non-destructive testing is essential for process development, monitoring, and quality assessment of CMC parts. Air-coupled ultrasound (ACU) is a fast and cost-efficient tool for non-destructive inspections of large components with respect to the detection of material inhomogeneities. Even though ACU inspection is usually used for visual inspection, the interpretation of C-scan images is often ambiguous with regard to critical defects and their impact on local material properties. This paper reports on a new approach to link the local acoustic damping of an oxide CMC plate obtained from ACU analysis with subsequent destructive mechanical testing and microstructural analyses. Local damping values of bending bars are extracted from ACU maps and compared with the results of subsequent resonant frequency damping analysis and 3-point bending tests. To support data interpretation, the homogeneous and inhomogeneous CMC areas detected in the ACU map are further analyzed by X-ray computed tomography and scanning electron microscopy. The results provide strong evidence that specific material properties such as Young’s modulus are not predictable from ACU damping maps. However, ACU shows a high, beneficial sensitivity for narrow but large area matrix cracks or delaminations, i.e., local damping is significantly correlated with specific properties such as shear moduli and bending strengths. Full article

We report here the synthesis and structural characterization of novel cationic (phenothiazinyl)vinyl-pyridinium (PVP) dyes, together with optical (absorption/emission) properties and their potential applicability as fluorescent labels. Convective heating, ultrasound irradiation and mechanochemical synthesis were considered as alternative synthetic methodologies proficient for overcoming drawbacks such as long reaction time, nonsatisfactory yields or solvent requirements in the synthesis of novel dye (E)-1-(3-chloropropyl)-4-(2-(10-methyl-10H-phenothiazin-3-yl)vinyl)pyridin-1-ium bromide 3d and its N-alkyl-2-methylpyridinium precursor 1c. The trans geometry of the newly synthesized (E)-4-(2-(7-bromo-10-ethyl-10H-phenothiazin-3-yl)vinyl)-1-methylpyridin-1-ium iodide 3b and (E)-1-methyl-4-(2-(10-methyl-10H-phenothiazin-3-yl)vinyl)pyridin-1-ium tetrafluoroborate 3a′ was confirmed by single crystal X-ray diffraction. A negative solvatochromism of the dyes in polar solvents was highlighted by UV-Vis spectroscopy and explanatory insights were supported by molecular modeling which suggested a better stabilization of the lowest unoccupied molecular orbitals (LUMO). The photostability of the dye 3b was investigated by irradiation at 365 nm in different solvents, while the steady-state and time-resolved fluorescence properties of dye 3b and 3a′ in solid state were evaluated under one-photon excitation at 485 nm. The in vitro cytotoxicity of the new PVP dyes on B16-F10 melanoma cells was evaluated by WST-1 assay, while their intracellular localization was assessed by epi-fluorescence conventional microscopy imaging as well as one- and two-photon excited confocal fluorescence lifetime imaging microscopy (FLIM). PVP dyes displayed low cytotoxicity, good internalization inside melanoma cells and intense fluorescence emission inside the B16-F10 murine melanoma cells, making them suitable staining agents for imaging applications. Full article

Proprotein convertases (PCSKs) process matrix metalloproteases and cytokines, but their function in the vasculature is largely unknown. Previously, we demonstrated upregulation of PCSK6 in atherosclerotic plaques from symptomatic patients, localization to smooth muscle cells (SMCs) in the fibrous cap and positive correlations with inflammation, extracellular matrix remodeling and cytokines. Here, we hypothesize that PCSK6 could be involved in flow-mediated vascular remodeling and aim to evaluate its role in the physiology of this process using knockout mice. Pcsk6^−/− and wild type mice were randomized into control and increased blood flow groups and induced in the right common carotid artery (CCA) by ligation of the left CCA. The animals underwent repeated ultrasound biomicroscopy (UBM) examinations followed by euthanization with subsequent evaluation using wire myography, transmission electron microscopy or histology. The Pcsk6^−/− mice displayed a flow-mediated increase in lumen circumference over time, assessed with UBM. Wire myography revealed differences in the flow-mediated remodeling response detected as an increase in lumen circumference at optimal stretch with concomitant reduction in active tension. Furthermore, a flow-mediated reduction in expression of SMC contractile markers SMA, MYH11 and LMOD1 was seen in the Pcsk6^−/− media. Absence of PCSK6 increases outward remodeling and reduces medial contractility in response to increased blood flow. Full article