Search Results (33)

Extracellular vesicles (EVs) are membrane-bound particles secreted by most cell types that play a pivotal role in intercellular communication via transporting protein, nucleic acid, lipid, and metabolite cargos. Among EVs, exosomes are a well-characterized subtype, typically ranging from 10–150 nm in diameter and originating from the endosomal pathway via the formation of multivesicular bodies that fuse with the plasma membrane. EVs/exosomes can be isolated from various biological fluids and cultured cells, with production and yield influenced by the cell type and culture conditions. Isolation methods, including ultracentrifugation or density-based ultracentrifugation, tangential flow filtration, size-exclusion chromatography, immunoaffinity and membrane-affinity capture, and recently developed commercial equipment, offer distinct advantages and limitations in terms of purity, scalability, and exosome integrity. Characterization techniques, such as nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), cryogenic electron microscopy (cryo-EM), atomic force microscopy (AFM), Western blotting, flow cytometry, and dynamic light scattering (DLS), assess exosome size, morphology, and biomarker expression. Given their biocompatibility and inherent targeting capabilities across a diverse range of diseases, EVs/exosomes hold clinical promise as diagnostic biomarkers, cell-free therapeutics, drug delivery vehicles, immune modulators, and in regenerative medicine. However, these emerging fields in exosome medicine continue to face challenges in standardizing EV sourcing, production, purification, yield, bio-targeting, drug loading, and drug delivery. While EVs/exosomes represent a rapidly advancing frontier in biomedical science, robust protocols for standardization and scalable production will be essential for their successful translation into clinical applications. This article provides a comprehensive overview of EV/exosome origins, their biological functions, the approaches for their isolation and characterization, and their therapeutic potential. Full article

The importance of protein-based materials in agricultural pest control has received increasing attention in recent years. Herein, Plutella xylostella brush-border membrane vesicles (BBMVs) were used as a target to screen for human domain antibodies with insecticidal activity. Three rounds of panning of the phage display library yielded the domain antibody C4D, which competed with the Cry1Ac toxin to bind to P. xylostella BBMVs. Against P. xylostella larvae, the recombinant soluble C4D protein showed an LC₅₀ of 1.57 μg/cm² (95% fiducial limits: 0.83–2.54). Using pull-down assays and liquid chromatography–tandem mass spectrometry, we identified the C4D binding partner in P. xylostella midgut BBMVs to be a cadherin-like protein. Bio-Layer Interferometry assay revealed that the dissociation constant between soluble C4D and P. xylostella cadherin-like protein was 2.99 × 10⁻⁶ M. Thus, the present study explored strategies to generate insecticidal antibodies, and the human domain antibody C4D identified and characterized in this study can serve as a framework for generating novel insecticidal agents. Full article

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and hold significant potential as biomarkers. Saliva, a non-invasive and easily accessible biofluid, offers a promising alternative to blood for miRNA-based diagnostics. However, miRNA profiling by next-generation sequencing (NGS) is highly influenced by library preparation protocol, which can introduce detection and quantification biases. This study compared four commercial small RNA library preparation kits—QIASeq miRNA library kit (Qiagen), RealSeq-Biofluids Plasma/Serum miRNA library kit (Somagenics), Small RNA-seq library prep kit (Lexogen) and NEBNext multiplex small RNA library prep set for illumina (set 1) (New England BioLabs)—to evaluate their performance in profiling miRNAs from cell-free saliva, plasma and their extracellular vesicles (EVs). Using both synthetic reference and biological samples, we assessed the kits’ efficiency in handling low RNA input, minimizing bias and detecting diverse miRNAs. QIAseq outperformed the others, showing the highest miRNA mapping rates, minimal adapter dimers and the broadest miRNA detection, particularly in saliva. Moreover, substantial overlap between saliva- and plasma-derived miRNAs supports saliva’s diagnostic potential. Overall, this study underscores the critical impact of library preparation on miRNA sequencing outcomes and offers guidance for selecting optimal protocols for biomarker discovery from non-invasive sample matrices. Full article

Raman spectroscopy has become an important tool for biomedical analysis due to its ability to provide label-free, non-destructive molecular fingerprints of biological samples. However, existing deep learning approaches for classifying biological Raman spectra often focus on specific datasets and lack generalizability and interpretability. In this study, BioRamanNet is presented, an interpretable and generalizable deep learning framework designed for classifying a wide range of biological Raman spectra. The model integrates adaptive one-dimensional convolutional layers and squeeze-and-excitation (SE) blocks within a residual network architecture to enhance feature extraction. BioRamanNet was evaluated using four representative Raman spectral datasets—breast cells, extracellular vesicles and particles (EVPs), viruses, and bacteria—achieving classification accuracies of 99.5%, 100%, 99.8%, and 85.3%, respectively. To improve model interpretability, a perturbation-based analysis using Voigt noise was introduced to identify key wavenumber regions influencing classification. These regions were found to correspond closely with known Raman biomarkers, validating their biological significance. The results of this work demonstrate that BioRamanNet is a powerful and interpretable tool for analyzing diverse biological Raman spectra and holds promise for advancing machine learning-assisted biomedical diagnostics. Full article

Exosomes, naturally derived extracellular vesicles, have emerged as powerful bio-nanocarriers in precision medicine. Their endogenous origin, biocompatibility, and ability to encapsulate and deliver diverse therapeutic payloads position them as transformative tools in drug delivery, gene therapy, and regenerative medicine. This review presents a comprehensive analysis of exosome-based therapeutics across multiple biomedical domains, including cancer, neurological and infectious diseases, immune modulation, and tissue repair. Exosomes derived from stem cells, immune cells, or engineered lines can be loaded with small molecules, RNA, or CRISPR-Cas systems, offering highly specific and low-immunogenic alternatives to viral vectors or synthetic nanoparticles. We explore endogenous and exogenous loading strategies, surface functionalization techniques for targeted delivery, and innovations that allow exosomes to traverse physiological barriers such as the blood–brain barrier. Furthermore, exosomes demonstrate immunomodulatory and regenerative properties in autoimmune and degenerative conditions, with promising roles in skin rejuvenation and cosmeceuticals. Despite their potential, challenges remain in large-scale production, cargo loading efficiency, and regulatory translation. Recent clinical trials and industry efforts underscore the accelerating momentum in this field. Exosomes represent a promising platform in precision medicine, though further standardization and validation are required before widespread clinical use. This review offers critical insights into current technologies, therapeutic mechanisms, and future directions to unlock the full translational potential of exosomes in clinical practice. Full article

In organic synthesis, the solvent is the chemical compound that represents the largest proportion of the process. However, conventional solvents are often toxic and dangerous for the environment, and an interesting alternative is to replace them by water. In this context, catalyst surfactants allow both organic reagents in water to be solubilized and organic reactions to be catalyzed. This article describes the synthesis of new biobased organocatalytic surfactants soluble in water, composed of a hydrocarbon chain grafted onto an imidazolidinone moiety. The imidazolidinone moiety acts as catalyst, but also as the polar head of the surfactant, while the fatty chain constitutes the hydrophobic tail. The five steps of the synthesis were optimized, respecting the principles of green chemistry, and two organocatalytic surfactants were obtained with a good selectivity. Surface properties in an aqueous medium were then evaluated with the use of tensiometric analysis. Their molecular organization in vesicles was characterized by Dynamic Light Scattering. The presence of vesicles allows reactions to be carried out in an organized aqueous medium. Model catalytic reactions performed in aqueous medium validated the feasibility of replacing conventional hazardous organic solvents. The newly synthesized biobased surfactants showed satisfactory catalytic activity and allowed the expected products to be obtained with good enantioselectivity. Full article

Several studies have reported the successful use of bio-orthogonal catalyst nanoparticles (NPs) for cancer therapy. However, the delivery of the catalysts to the target tissues in vivo remains an unsolved challenge. The combination of catalytic NPs with extracellular vesicles (EVs) has been proposed as a promising approach to improve the delivery of therapeutic nanomaterials to the desired organs. In this study, we have developed a nanoscale bio-hybrid vector using a CO-mediated reduction at low temperature to generate ultrathin catalytic Pd nanosheets (PdNSs) as catalysts directly inside cancer-derived EVs. We have also compared their biodistribution with that of PEGylated PdNSs delivered by the EPR effect. Our results indicate that the accumulation of PdNSs in the tumour tissue was significantly higher when they were administered within the EVs compared to the PEGylated PdNSs. Conversely, the amount of Pd found in non-target organs (i.e., liver) was lowered. Once the Pd-based catalytic EVs were accumulated in the tumours, they enabled the activation of a paclitaxel prodrug demonstrating their ability to carry out bio-orthogonal uncaging chemistries in vivo for cancer therapy. Full article

In recent years, unidimensional photonic crystal-based biosensors have attracted significant attention due to their potential for label-free bio-detection of cells, proteins, and other organic molecules. These biosensors are based on alternating materials with different refractive indices and a cavity region in which biomolecules can be injected. In this study, we investigated numerically the optical properties of unidimensional photonic crystals based on [LiTaO₃/MgF₂], [LiTaO₃/SiO₂], [PbTiO₃/MgF₂], and [PbTiO₃/SiO₂] heterostructures, and focused our discussion on the detection of four kinds of extracellular vesicles. Our results demonstrated that the [PbTiO₃/MgF₂] photonic crystal biosensor exhibited the best biosensing performance, with a maximum value of the sensitivity of 511.3 nm/RIU. This study provides valuable insights into the use of perovskites materials, in particular, LiTaO₃ and PbTiO₃ for photonic crystal-based biosensors for various applications, including disease diagnosis and monitoring therapy responses. Full article

Aging is a complex, multifaceted degenerative process characterized by a progressive accumulation of macroscopic and microscopic modifications that cause a gradual decline of physiological functions. During the last few years, strategies to ease and counteract senescence or even rejuvenate cells and tissues were proposed. Here we investigate whether young cell secretome-derived extracellular vesicles (EVs) ameliorate the cellular and physiological hallmarks of aging in senescent cells. In addition, based on the assumption that extracellular matrix (ECM) provides biomechanical stimuli, directly influencing cell behavior, we examine whether ECM-based bio-scaffolds, obtained from decellularized ovaries of young swine, stably maintain the rejuvenated phenotype acquired by cells after exposure to young cell secretome. The results obtained demonstrate that young cells release EVs endowed with the ability to counteract aging. In addition, comparison between young and aged cell secretomes shows a significantly higher miR-200 content in EVs produced using fibroblasts isolated from young donors. The effect exerted by young cell secretome-derived EVs is transient, but can be stabilized using a young ECM microenvironment. This finding indicates a synergistic interaction occurring among molecular effectors and ECM-derived stimuli that cooperate to control a unique program, driving the cell clock. The model described in this paper may represent a useful tool to finely dissect the complex regulations and multiple biochemical and biomechanical cues driving cellular biological age. Full article

Cancers are defined by genetic defects, which underlines the prospect of using gene therapy in patient care. During the past decade, CRISPR technology has rapidly evolved into a powerful gene editing tool with high fidelity and precision. However, one of the impediments slowing down the clinical translation of CRISPR-based gene therapy concerns the lack of ideal delivery vectors. Extracellular vesicles (EVs) are nano-sized membrane sacs naturally released from nearly all types of cells. Although EVs are secreted for bio-information conveyance among cells or tissues, they have been recognized as superior vectors for drug or gene delivery. Recently, emerging evidence has spotlighted EVs in CRISPR delivery towards cancer treatment. In this review, we briefly introduce the biology and function of the CRISPR system and follow this with a summary of current delivery methods for CRISPR applications. We emphasize the recent progress in EV-mediated CRISPR editing for various cancer types and target genes. The reported strategies for constructing EV-CRISPR vectors, as well as their limitations, are discussed in detail. The review aims to throw light on the clinical potential of engineered EVs and encourage the expansion of our available toolkit to defeat cancer. Full article

Exosomes are a subpopulation of extravascular vesicles with a diameter of 30–150 nm. They are cellular-communication mediators, often reaching very distant organism tissues. Information is transferred by exosomal cargo, composed of a wide variety of macromolecules such as nucleic acids, proteins, and lipids. Exosomes possess natural specific cell targeting properties that are desirable in designing targeted macromolecules (DNA and RNA) and drug delivery systems (doxorubicin, paclitaxel, and taxol). In this context, exosomes can be defined as bio-derived drug transporting and protecting devices for the treatment of bacterial (toxoplasmosis and salmonellosis), viral (AIDS and hepatitis B), and cancer (lung, pancreatic, colon, brain, and breast) diseases. Extensive research proves that exosomes’ natural cargo can double-act, both increasing and decreasing the disease severity. In this case, the exosomes need to be prepared, namely, their origin and their cargo need to be screened and known. Thus, appropriate methods for intact and price-effective exosome isolation are needed with further exosome properties description. Among many utilized isolation methods, the most common are ultracentrifugation, polymer-based precipitation, and affinity precipitation-isolation systems, but novel microfluidic methods compromising high efficacy and purity are being developed. In this review, we state the current knowledge and trends in exosome-based drug delivery systems. Full article

Cardiac tissue engineering has emerged as a promising strategy to treat infarcted cardiac tissues by replacing the injured region with an ex vivo fabricated functional cardiac patch. Nevertheless, integration of the transplanted patch with the host tissue is still a burden, limiting its clinical application. Here, a bi-functional, 3D bio-printed cardiac patch (CP) design is proposed, composed of a cell-laden compartment at its core and an extracellular vesicle (EV)-laden compartment at its shell for better integration of the CP with the host tissue. Alginate-based bioink solutions were developed for each compartment and characterized rheologically, examined for printability and their effect on residing cells or EVs. The resulting 3D bio-printed CP was examined for its mechanical stiffness, showing an elastic modulus between 4–5 kPa at day 1 post-printing, suitable for transplantation. Affinity binding of EVs to alginate sulfate (AlgS) was validated, exhibiting dissociation constant values similar to those of EVs with heparin. The incorporation of AlgS-EVs complexes within the shell bioink sustained EV release from the CP, with 88% cumulative release compared with 92% without AlgS by day 4. AlgS also prolonged the release profile by an additional 2 days, lasting 11 days overall. This CP design comprises great potential at promoting more efficient patch assimilation with the host. Full article

Insulin replacement is an available treatment for autoimmune type 1 diabetes mellitus (T1DM). There are multiple limitations in the treatment of autoimmune diseases such as T1DM by immunosuppression using drugs and chemicals. The advent of extracellular vesicle (EV)-based therapies for the treatment of various diseases has attracted much attention to the field of bio-nanomedicine. Tolerogenic nanoparticles can induce immune tolerance, especially in autoimmune diseases. EVs can deliver cargo to specific cells without restrictions. Accordingly, EVs can be used to deliver tolerogenic nanoparticles, including iron oxide-peptide-major histocompatibility complex, polyethylene glycol-silver-2-(1′H-indole-3′-carbonyl)-thiazole-4-carboxylic acid methyl ester, and carboxylated poly (lactic-co-glycolic acid) nanoparticles coupled with or encapsulating an antigen, to effectively treat autoimmune T1DM. The present work highlights the advances in exosome-based delivery of tolerogenic nanoparticles for the treatment of autoimmune T1DM. Full article

Exosomes are lipid-bilayer enclosed vesicles with diameters of 30–150 nm, which play a pivotal role in cell communication by transporting their cargoes such as proteins, lipids, and genetic materials. In recent years, exosomes have been under intense investigation, as they show great promise in numerous areas, especially as bio-markers in liquid biopsies. However, due to the high heterogeneity and the nano size of exosomes, the separation of exosomes is not easy. This review will deliver an outline of the conventional methods and the microfluidic-based technologies for exosome separation. Particular attention is devoted to microfluidic devices, highlighting the efficiency of exosome isolation by these methods. Additionally, this review will introduce advances made in the integrated microfluidics technologies that enable the separation and analysis of exosomes. Full article

The self-assembly of lipid mixtures in aqueous solution was investigated by dissipative particle dynamics simulation. Two types of lipid molecules were modelled, where three mixed structures, i.e., the membrane, perforated membrane and vesicle, were determined in the self-assembly processes. Phase behaviour was investigated by using the phase diagrams based on the tail chain lengths for the two types of lipids. Several parameters, such as chain number and average radius of gyration, were employed to explore the structural formations of the membrane and perforated membrane in the dynamic processes. Interface tension was used to demonstrate the mechanical properties of the membrane and perforated membrane in the equilibrium state and dynamics processes. Results help us to understand the self-assembly mechanism of the biomolecule mixtures, which has a potential application for designing the lipid molecule-based bio-membranes in solutions. Full article