Introduction:Early identification of tumor-like changes in bone remains a major diagnostic challenge because conventional imaging methods mainly rely on structural contrast and may involve ionizing radiation, high cost, or limited point-of-care accessibility. In line with the growing interest in bioelectronic and nanoelectronic biosensor technologies for translational diagnostics, this study investigates a low-cost, non-ionizing microwave sensing platform for detecting dielectric changes associated with a tumor-mimicking bone phantom. Methods:A compact microstrip resonator antenna was designed in CST Microwave Suite and fabricated on an FR4 substrate for localized near-field sensing. A multilayer bone phantom containing cortical bone- and marrow-mimicking regions was prepared using wheat flour, deionized water, dextrose, and olive oil. A separate water-rich gelatin-based tumor phantom was prepared to reproduce the higher dielectric response expected from malignant tissue. The dielectric properties of the bone and tumor phantoms were measured using a Vector Network Analyzer (VNA)-based open-ended coaxial probe system. The fabricated antenna was then experimentally evaluated by reflection coefficient (S11) measurements at two healthy phantom positions and one tumor-over-phantom position. Results:Dielectric characterization confirmed a clear contrast between the bone phantom and tumor-like region in both the real and imaginary parts of relative permittivity. The tumor-loaded configuration produced a downward resonance shift of 110 MHz around 4.2-4.3 GHz, relative to the healthy reference, which was 2.75 times larger than the variation between the two healthy positions. In addition, a 4.47 dB change in S11 magnitude was observed, confirming that the tumor-like inclusion altered the near-field dielectric loading and impedance-matching condition of the resonator. Conclusions:The proposed microstrip resonator antenna demonstrates the feasibility of a compact RF bioelectronic sensing approach for detecting tumor-like dielectric perturbations in a controlled bone phantom environment. Although not intended as a clinical diagnostic device at this stage, the results support further development of this platform toward translational microwave biosensing, more realistic preclinical phantoms, array-based configurations, and AI-assisted classification for future diagnostic applications.
Theranostic technologies that incorporate nanotechnology into ultrasound (US) and photoacoustic imaging (PAI) show great promise for the diagnosis and treatment of a wide range of diseases. Despite recent advancements in nanomaterials, US/PA systems, which use both endogenous and exogenous contrast agents, are now more sensitive and effective in therapy. Intrinsic optical absorption-based label-free imaging is made possible by endogenous agents such as collagen, lipids, haemoglobin, and melanin. Exogenous contrast agents, on the other hand, are engineered to circumvent drawbacks such as low tissue selectivity and weak signal intensity. Advanced multimodal theranostic applications are made possible by these designed nanomaterials, which improve optical absorption, acoustic responsiveness, and targeted imaging. The benefits of optical high sensitivity and acoustic deep penetration are brought together in PAI, an integrated biomedical imaging modality. It may produce structural and functional pictures with enough contrast and resolution, providing a wealth of pathology data for disease-oriented diagnostics. Consequently, it has become an effective tool of precision nanomedicine and has discovered numerous uses so far. PAI is a hybrid imaging method that uses both optical and acoustic imaging to produce detailed pictures with a wealth of information about multiple parameters. This review explains the underlying principle of US/PAI bimodal imaging techniques, including their technical benefits, contrast agents, and biomedical applications. Additionally, we go over the difficulties and potential applications of US/PAI bimodal imaging in clinical settings.
Background:CLEC5A (C-type lectin domain family 5 member A) is an innate immune receptor implicated in inflammatory signaling, contributing to hyperinflammatory responses in infections and sterile inflammation. However, CLEC5A dynamics in human diseases remain to be identified. Here, we systematically characterized CLEC5A dynamics in humans across cells, tissues, and disease states, and to explore the functional significance of CLEC5A in macrophage activation based on single-cell genomics. Methods:With multi-omics (scRNA-seq, proteomics and big data analytics), we analyzed extensive human transcriptomic datasets (>42,000 samples) to profile CLEC5A expression by cell type, tissue, and disease. Single-nucleus RNA-seq (snRNA-seq) from pediatric congenital heart disease and a virtual CLEC5A gene knockout were also performed to characterize CLEC5A dynamics in humans. Results:CLEC5A is highly enriched in innate immune cells, particularly in macrophages and neutrophils. Baseline CLEC5A in most tissues is low, but it is markedly upregulated in inflammatory and infectious diseases. CLEC5A expression has sex-specific differences in certain organs. Single-cell analysis showed that CLEC5A can be considered novel marker of proinflammatory macrophages with elevated cytokine production, antigen presentation, and impaired phagocytosis. Virtual CLEC5A knockout analysis identified coordinated perturbation of immune-regulatory pathways and overlapping genes linking CLEC5A to macrophage activation networks. Conclusion:CLEC5A is predominantly expressed in myeloid cells and acts as a key amplifier of inflammation in human diseases. Our findings highlight CLEC5A as a potential biomarker and therapeutic target in myeloid-driven hyperinflammatory conditions, warranting further experimental and translational validation.
The tumor microenvironment (TME) is a dynamic and heterogeneous niche that critically shapes cancer progression, immune evasion, and therapeutic resistance. Characterized by gradients in oxygen tension, pH, and metabolic activity, the TME offers a rich yet underexploited source of real-time biomarkers related to cancer. While conventional imaging techniques often lack the temporal resolution and molecular specificity to capture these rapid physiological changes, emerging miniaturized bioelectronics and multiplexed systems carry promise for in situ monitoring of such TME markers with high sensitivity and spatial precision. This article explores such recent advances in this direction, including bioelectronic sensor design, flexible electrochemical devices, organic transistors, and nanostructured interfaces tailored for TME characterization. Further, a discussion of the convergence of bioelectronics with nano-contrast-based molecular imaging is presented, with prospects for developing closed-loop therapeutic systems for cancer. These technologies offer a transformative platform for precision oncology, enabling dynamic, continuous, and localized assessment of tumor biology across preclinical and clinical settings.
The use of raw bovine milk as a source of extracellular vesicles (EVs) has gained in interest for therapeutic applications due to its low cost, accessibility, low immunogenicity, and potential for oral delivery. To address the need to achieve higher throughput isolation of high-quality EVs, high performance liquid chromatography (HPLC) using a hydrophobic interaction chromatography (HIC) capture/elution program is performed on microbore, capillary-channeled polymer (C-CP) fiber columns. Methods: Bovine milk is first skimmed to reduce the fat-laden matrix, followed by treatment using acetic acid (Ac) to precipitate casein micelles before isolation of milk-derived EVs (MDEVs) via HPLC using an HIC process modality with PET C-CP fiber columns. The treated milk is introduced to the column under EV binding conditions, where an ionic solvent with a small amount of organic modifier causes salts, small molecules, and proteinaceous species to pass through unretained while retaining the EVs on-column. The target EVs are eluted by decreasing the ionic strength of the solvent and increasing the elution strength. Results: EVs isolated using PET C-CP fiber columns demonstrate the removal of >95% of matrix-related concomitant species and yield particle densities on the order of 1011 particles mL-1 in 20 min. Validation of the success of the separation is demonstrated through response curves, nanoflow cytometry, transmission electron microscopy, and protein assays in accordance with MISEV guidelines. Conclusions: A rapid approach to the high yield isolation of high-quality MDEVs via microbore-scale PET C-CP fiber columns is presented here. Each separation yields MDEVs on the order of 4 x 1011 particles mL-1, in 20-min at a cost of <$5 per column. Paths forward to greater EV throughput and yields are currently under development.
One of the major barriers to further clinical adoption of photoacoustic imaging (PAI) and photothermal therapy (PTT) is a lack of versatile and multi-faceted contrast agents. While indocyanine green (IcG) has gained considerable attention as a promising contrast agent, it is severely limited by rapid clearance time, low photostability, and a peak absorbance wavelength that coincides with hemoglobin. Herein, Liposomal J-aggregates of Indocyanine green (LJA) are presented as a superior alternative to generic IcG. They are a facilely produced, biodegradable nanoparticle offering advantageous physiochemical properties. J-aggregation results in a redshifted peak absorbance of increased magnitude and higher photostability compared to IcG. Liposome encapsulation increases circulation time leading to improved tumor uptake. Monte Carlo modeling of light interaction with tissue suggests that these improved optical properties make LJA a better contrast agent for PTT not only at longer wavelengths like 852nm and 890nm, but also at the commonly available and widely used 808nm. In vitro and in vivo testing support first approximations from modelling as LJA provide significantly higher photoacoustic signal, show increased tumor uptake, reach significantly higher temperatures under photothermal irradiation, and have the capacity to reduce tumor growth following therapy.
Kidney inflammation is a central driver of acute kidney injury (AKI) and its progression to chronic kidney disease (CKD). While several imaging and biomarker-based approaches are under development, clinically validated non-invasive methods to directly quantify renal inflammation remain limited. This study introduces a novel approach using contrast-enhanced ultrasound (CEUS) with Cy5-labeled nanobubbles (NBs) to address this critical knowledge gap. Using a murine ischemia-reperfusion injury (IRI) model, CEUS imaging enabled real-time visualization of inflammation-induced changes in kidney perfusion and vascular integrity. Parametric analyses of non-linear imaging revealed delayed time-to-peak (TTP) and increased area under the falling curve (AUfC) in IRI kidneys, suggesting impaired microvascular perfusion and NB retention. Decorrelation time (DT) mapping further identified prolonged NB retention in the IRI group, indicating increased capillary permeability and NB extravasation. These findings correlated with histological and immunofluorescent analyses, which confirmed the presence of tubular injury, extravascular Cy5 signal localization, and increased neutrophil infiltration in inflamed kidney tissues. This study is the first to establish CEUS with NBs as a non-invasive, quantitative method for measuring kidney inflammation. With strong correlations between imaging metrics and histologic injury scores, this technology provides an accessible and non-invasive tool for monitoring renal inflammation and reducing reliance on invasive renal biopsies.
Introduction: Bovine amniotic membrane (BAM) and secretome are tissue engineering materials studied for their high healing effects. This study aims to evaluate the expression levels of BMP-2, OPG, RANKL, and osteoblast count following the administration of BAM combined with secretome in Wistar rats undergoing socket preservation procedures. Method: This study is a pure experimental study with a randomised post-test only control group design. Pre-test measurements were not performed because the baseline condition of bone healing cannot be assessed prior to extraction without interfering with socket healing. Randomization ensured that all groups started with equivalent baseline conditions, allowing reliable comparison of outcomes across groups. The expression of BMP-2, OPG, RANKL, and the number of osteoblasts in the dental tissues of male Wistar rats were examined post-extraction following socket preservation. The research variables consisted of control, as well as those treated with BAM, secretome, and BAM-secretome. One-Way ANOVA analysis and Tukey's Post-Hoc test were conducted to compare the expression of dependent variables between treatment groups. Results: One-Way ANOVA and Post-Hoc Tukey tests showed that the expression of BMP-2, OPG, RANKL, and the number of osteoblasts in the BAM-secretome group were significantly higher compared to control, BAM, and secretome (p<0.001). Conclusion: The combination of BAM with secretome significantly enhanced the expression of BMP-2, OPG, and osteoblast counts compared to BAM, secretome alone, and control groups with no treatment. These findings suggest that BAM-secretome holds promising potential for promoting bone regeneration. However, further phased clinical trials are essential to evaluate its safety and efficacy in socket preservation treatments.
Prolonged three-dimensional culture exposes stem cells to sustain microenvironmental and mechanical stresses that can promote aging- and senescence-associated phenotypic alterations. This study examined how long-term expansion of human bone marrow-derived mesenchymal stem cells (BMSCs) in a hollow fiber bioreactor (HFB) influences cellular senescence and the molecular composition of secreted small extracellular vesicles (sEVs). During extended HFB culture, BMSCs exhibited progressive morphological flattening and cytoskeletal disorganization, accompanied by increased senescence-associated β-galactosidase activity and immunophenotypic remodeling characterized by reduced fluorescence intensity and spatial redistribution of canonical MSC markers, consistent with a stress-adapted, early senescence-associated cellular state. In parallel, sEVs were collected longitudinally over 40 days and characterized by nanoparticle tracking analysis, immunoblotting, and quantitative proteomics. While vesicle size, marker expression, and yield remained stable throughout culture, proteomic profiling revealed pronounced, phase-dependent remodeling of sEV cargo, including coordinated alterations in oxidative stress-related processes, lysosomal and extracellular matrix-associated pathways, and relative depletion of cytoskeletal and translational components. Notably, these vesicular signatures closely mirrored senescence-associated changes observed at the cellular level. The strong correspondence between cellular phenotypes and sEV proteomic profiles establishes vesicle analysis as a convergent and noninvasive readout of BMSC aging, enabling sensitive monitoring of senescence progression while reducing reliance on parallel, labor-intensive cellular assays. Collectively, these findings indicate that prolonged HFB culture promotes a controlled, stress-associated senescence program in BMSCs and position sEV proteomic profiling as a robust approach for assessing stem cell aging dynamics during long-term three-dimensional bioreactor culture.
