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Targeting the delivery of anti-cancer drugs to a tumor site is essential for effective treatment and to ensure minimal damage to healthy cells and tissues. In this work, a chitosan-based nanoplatform was constructed for combined photothermal therapy and chemotherapy of breast cancer. The pH-sensitive and biocompatible biopolymer chitosan (CS) was grafted with N-vinylcaprolactam (NVCL) and modified with biotin (Bio), imparting it with temperature sensitive property and also the ability for active targeting. The polymer self-assembled to give nanoparticles (NPs) loaded with indocyanine green (ICG) and doxorubicin (DOX). When the NPs are exposed to near-infrared (NIR) laser irradiation, ICG converts the light to heat, inducing a significant phase transition in the NPs and facilitating the release of the drug cargo. In addition, the solubility of chitosan is increased in the slightly acidic microenvironment of the tumor site, which also promotes drug release. A detailed analysis of the NPs both in vitro and in vivo showed that the carrier system is biocompatible, while the drug-loaded NPs are selectively taken up by cancer cells. Particularly when augmented with NIR irradiation, this leads to potent cell death in vitro and also in an in vivo murine xenograft model of breast cancer.

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Despite recent advances in the field of mRNA therapy, the lack of safe and efficacious delivery vehicles with pharmaceutically developable properties remains a major limitation. Here, we describe the systematic optimisation of lipid-peptide nanocomplexes for the delivery of mRNA in two murine cancer cell types, B16-F10 melanoma and CT26 colon carcinoma as well as NCI-H358 human lung bronchoalveolar cells. Different combinations of lipids and peptides were screened from an original lipid-peptide nanocomplex formulation for improved luciferase mRNA transfection in vitro by a multi-factorial screening approach. This led to the identification of key structural elements within the nanocomplex associated with substantial improvements in mRNA transfection efficiency included alkyl tail length of the cationic lipid, the fusogenic phospholipid, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and cholesterol. The peptide component (K

Coronary heart disease is a leading cause of death worldwide. Paramedics are instrumental in the treatment, triage and transport of patients experiencing acute coronary syndromes and acute myocardial infarction (AMI). Paramedics currently rely on prehospital electrocardiography and patient symptomology to diagnose AMI, which may result in missed diagnoses. Point-of-care biomarkers such as cardiac troponin (cTnT) and copeptin may have the potential to increase the diagnostic capabilities of prehospital providers. Multiple electronic databases (MAG Online Library, Cochrane Library, PubMed and Embase) were searched to evaluate the feasibility and potential application of cardiac biomarkers cTnT and copeptin in the prehospital setting. Because of the paucity of evidence, this article explores the evidence on the dual-marker strategy of cTnT and copeptin to increase diagnostic capabilities of prehospital providers, and as an adjunct for decision-making and risk stratification for AMI. The evidence suggests that using the combined dual-marker strategy of cTnT and copeptin may counteract limitations of the ‘troponin-blind’ period of cTnT and the low cardiac specificity of copeptin. However, the research for this method is still in its infancy and requires investigation into its feasibility and affordability as well as into the training required to implement it in paramedic practice.

In this study, gadolinium (Gd) doped MgAl layered double hydroxides (LDHs) were synthesized via a ‘bottom-up’ method and fully characterized by X-ray diffraction, infrared spectroscopy and relaxivity measurements. Two cytotoxic agents were then intercalated via ion-exchange. X-ray diffraction patterns exhibit expanded interlayer spacings as a result of successful drug intercalation. Infrared spectra also showed characteristic peaks of the incorporated methotrexate (MTX) or 5-fluorouracil (5-FU). The LDHs were found to be highly stable under physiological conditions, while in acidic conditions a small proportion of Gd was freed into the immersion medium. Dissolution tests revealed that both 5FU and MTX were rapidly released from the LDH carrier. The longitudinal relaxivity of Gd-LDHs remains largely stable during drug release over 24 h, and was higher in acidic environments. Overall, the drug-loaded Gd-LDH systems prepared in this study could serve as pH-sensitive theranostic platforms for MRI-guided anti-cancer therapy.

Supplementary Data from Cdc7 Kinase Is a Predictor of Survival and a Novel Therapeutic Target in Epithelial Ovarian Carcinoma

Purpose: The DNA replication licensing machinery is integral to the control of proliferation, differentiation, and maintenance of genomic stability in human cells. We have analyzed replication licensing factors (RLF), together with DNA ploidy status, to investigate their role in progression of penile squamous cell carcinoma and to assess their utility as novel prognostic tools.Experimental Design: In a cohort of 141 patients, we linked protein expression profiles of the standard proliferation marker Ki67 and the RLFs Mcm2 and geminin to clinicopathologic variables, ploidy status, and clinical outcome.Results: Increased Ki67, Mcm2, and geminin levels were each significantly associated with arrested tumor differentiation (P < 0.0001) and aneuploidy (P ≤ 0.01). Accelerated cell cycle progression was linked to increasing tumor size, stage, and depth of invasion. Aneuploid tumors significantly correlated with tumor grade (P < 0.0001). Biomarker expression and DNA ploidy status were significant predictors of locoregional disease progression [Mcm2 (P = 0.02), geminin (P = 0.02), Ki67 (P = 0.03), and aneuploidy (P = 0.03)] in univariate analysis. Importantly, aneuploidy was a strong independent prognosticator for overall survival (hazard ratio, 4.19; 95% confidence interval, 1.17-14.95; P = 0.03). Used in conjunction with conventional pathologic information, multiparameter analysis of these variables can stratify patients into low- or high-risk groups for disease progression (Harrell's c-index = 0.88).Conclusions: Our findings suggest that RLFs and tumor aneuploidy may be used as an adjunct to conventional prognostic indicators, identifying men at high risk of disease progression. Our results also identify the DNA replication initiation pathway as a potentially attractive therapeutic target in penile squamous cell carcinoma. (Clin Cancer Res 2009;15(23):7335–44)

Purpose: There is a lack of prognostic and predictive biomarkers in epithelial ovarian carcinoma, and the targeting of oncogenic signaling pathways has had limited impact on patient survival in this highly heterogeneous disease. The origin licensing machinery, which renders chromosomes competent for DNA replication, acts as a convergence point for upstream signaling pathways. We tested the hypothesis that Cdc7 kinase, a core component of the licensing machinery, is predictive of clinical outcome and may constitute a novel therapeutic target in epithelial ovarian carcinoma.Experimental Design: A total of 143 cases of ovarian cancer and 5 cases of normal ovary were analyzed for Cdc7 protein expression dynamics and clinicopathologic features. To assess the therapeutic potential of Cdc7, expression was down-regulated by RNA interference in SKOV-3 and Caov-3 ovarian cancer cells.Results: Increased Cdc7 protein levels were significantly associated with arrested tumor differentiation (P = 0.004), advanced clinical stage (P = 0.01), genomic instability (P < 0.001), and accelerated cell cycle progression. Multivariate analysis shows that Cdc7 predicts disease-free survival independent of patient age, tumor grade and stage (hazard ratio, 2.03; confidence interval, 1.53-2.68; P < 0.001), with the hazard ratio for relapse increasing to 10.90 (confidence interval, 4.07-29.17) for the stages 3 to 4/upper Cdc7 tertile group relative to stages 1 to 2/lower Cdc7 tertile tumors. In SKOV-3 and Caov-3 cells, Cdc7 siRNA knockdown triggered high levels of apoptosis, whereas untransformed cells arrest in G1 phase and remain viable.Conclusions: Our findings show that Cdc7 kinase predicts survival and is a potent anticancer target in epithelial ovarian carcinoma, highlighting its potential as a predictor of susceptibility to small molecule kinase inhibitors currently in development.

Supplementary Data from DNA Replication Licensing Factors and Aneuploidy Are Linked to Tumor Cell Cycle State and Clinical Outcome in Penile Carcinoma

IntroductionExcessive screen exposure (ESE) is a growing global public health concern. This study aims to investigate the potential association between ESE and suspected developmental coordination disorder (DCD) in Chinese pre-schoolers, with or without siblings.MethodA retrospective cohort study was conducted, involving 126,433 children from 551 cities in China. The Little Developmental Coordination Disorder Questionnaire (LDCDQ) was employed to evaluate motor impairment in children, while parents provided information on their children’s screen time in the past year. A mixed and multi-level logistic regression model was used to analyze the associations of all screen exposure measurements from the past year with LDCDQ scores and the risk of suspected DCD.ResultsThe prevalence of excessive screen exposure was 67.6% (>1 h per day) and 28.9% (>2 h per day) in Chinese pre-schoolers. One hour’s increase in weekday daily screen time, weekend daily screen time, and screen time before sleep in the past year was associated with a decreased total score of the LDCDQ (β were −0.690, −0.398, and −1.587, p 1 vs. ≤1 h) and 1.162 (>2 vs. ≤2 h) and increased risks of suspected DCD by 44.0% (>1 vs. ≤1 h) and 31.1% (>2 vs. ≤2 h) with statistical significance (each p ConclusionThe risk of suspected DCD was highest for screen time exposure before bed compared with average weekday and weekend exposures. Parents should be advised to prevent their children from using electronic screens unsupervised, especially in one-child families.

Highly competitive coral reef benthic communities are acutely sensitive to changes in environmental parameters such as temperature and nutrient concentrations. Physical oceanographic processes that induce upwelling therefore act as drivers of community structure on tropical reefs. How upwelling impacts coral communities, however, is not fully understood; upwelling may provide a natural buffer against climate impacts and could potentially enhance the efficacy of spatial management and reef conservation efforts. This study employed a systematic review to assess existing literature linking upwelling with reef community structure, and a meta-analysis to quantify upwelling impact on the percentage cover of coral reef benthic groups. We show that upwelling has context-dependant effects on the cover of hard coral and fleshy macroalgae, with effect size and direction varying with depth, region and remoteness. Fleshy macroalgae were found to increase by 110% on inhabited reefs yet decrease by 56% around one well-studied remote island in response to upwelling. Hard coral cover was not significantly impacted by upwelling on inhabited reefs but increased by 150% when direct local human pressures were absent. By synthesizing existing evidence, this review facilitates adaptive and nuanced reef management which considers the influence of upwelling on reef assemblages.

Nano-zirconia (ZO) was synthesized using a microwave-assisted one-pot precipitation route. Two biopolymers, chitosan (CTS) and carboxymethyl cellulose were blended with ZO at different w/w ratios. The formulation with 30% w/w chitosan (ZO-CTS) was found to give enhanced uptake of F − and As(V). ZO and the most effective ZO-CTS system were characterized using Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and X-ray photoelectron spectroscopy. These confirmed the formation of a composite system containing nanoparticles of 50 nm in size, in which ZO was present in the amorphous form. It was observed that the combination of ZO with CTS improved the F − and As(V) adsorption capacity most notably at pH 5.5. Fluoride adsorption by ZO-CTS followed the Freundlich isotherm model, with an adsorption capacity of 120 mg g −1 . Adsorption of As(V) by ZO-CTS could be fitted with both the Langmuir and Freundlich isotherm models and was found to have a capacity of 14.8 mg g −1 . Gravity filtration studies conducted for groundwater levels indicated the effectiveness of ZO-CTS in adsorbing As(V) and F − at a pH of 5.5. The ability of the ZO-CTS in removing Cd(II) and Pb(II) was also investigated, and no such enhancement was observed, and found the neat ZO was the most potent sorbent here.

IntroductionPulmonary-resident memory T cells (TRM) and B cells (BRM) orchestrate protective immunity to reinfection with respiratory pathogens. Developing methods for the in situ detection of these populations would benefit both research and clinical settings.MethodsTo address this need, we developed a novel in situ immunolabelling approach combined with clinic-ready fibre-based optical endomicroscopy (OEM) to detect canonical markers of lymphocyte tissue residency in situ in human lungs undergoing ex vivo lung ventilation (EVLV).ResultsInitially, cells from human lung digests (confirmed to contain TRM/BRM populations using flow cytometry) were stained with CD69 and CD103/CD20 fluorescent antibodies and imaged in vitro using KronoScan, demonstrating it’s ability to detect antibody labelled cells. We next instilled these pre-labelled cells into human lungs undergoing EVLV and confirmed they could still be visualised using both fluorescence intensity and lifetime imaging against background lung architecture. Finally, we instilled fluorescent CD69 and CD103/CD20 antibodies directly into the lung and were able to detect TRM/BRM following in situ labelling within seconds of direct intra-alveolar delivery of microdoses of fluorescently labelled antibodies.DiscussionIn situ, no wash, immunolabelling with intra-alveolar OEM imaging is a novel methodology with the potential to expand the experimental utility of EVLV and pre-clinical models.

HIV nonprogression despite persistent viremia is rare among adults who are naive to antiretroviral therapy (ART) but relatively common among ART-naive children. Previous studies indicate that ARTnaive pediatric slow progressors (PSPs) adopt immune evasion strategies similar to those described in natural hosts of SIV. However, the mechanisms underlying this immunophenotype are not well understood. In a cohort of early-treated infants who underwent analytical treatment interruption (ATI) after 12 months of ART, expression of PD-1 on CD8+ T cells immediately before ATI was the main predictor of slow progression during ATI. PD-1+CD8+ T cell frequency was also negatively correlated with CCR5 and HLA-DR expression on CD4+ T cells and predicted stronger HIV-specific T lymphocyte responses. In the CD8+ T cell compartment of PSPs, we identified an enrichment of stem-like TCF-1+PD-1+ memory cells, whereas pediatric progressors and viremic adults had a terminally exhausted PD-1+CD39+ population. TCF-1+PD-1+ expression on CD8+ T cells was associated with higher proliferative activity and stronger Gag-specific effector functionality. These data prompted the hypothesis that the proliferative burst potential of stem-like HIV-specific cytotoxic cells could be exploited in therapeutic strategies to boost the antiviral response and facilitate remission in infants who received early ART with a preserved and nonexhausted T cell compartment.

Ataxia-telangiectasia mutated (ATM) drives the DNA damage response via modulation of multiple signal transduction and DNA repair pathways. Previously, ATM activity was implicated in promoting the non-homologous end joining (NHEJ) pathway to repair a subset of DNA double strand breaks (DSBs), but how ATM performs this function is still unclear. In this study, we identified that ATM phosphorylates the DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a core NHEJ factor, at its extreme C-terminus at threonine 4102 (T4102) in response to DSBs. Phosphorylation at T4102 stabilizes the interaction between DNA-PKcsand the Ku-DNA complex and promotes assembly and stabilization of the NHEJ machinery at DSBs. Ablating phosphorylation at this site results in decreased NHEJ, radiosensitivity, and increased radiation-induced genomic instability. Collectively, these findings establish a key role for ATM in NHEJ-dependent repair of DSBs through positive regulation of DNA-PKcs.

Extracellular vesicles (EVs) are routinely released from nearly all cell types as transport vehicles and for cell communication. Crucially, they contain biomolecular content for the identification of health and disease states that can be detected from readily accessible physiological fluids, including urine, plasma, or saliva. Despite their clinical utility within noninvasive diagnostic platforms such as liquid biopsies, the currently available portfolio of analytical approaches are challenged by EV heterogeneity in size and composition, as well as the complexity of native biofluids. Quartz crystal microbalance with dissipation monitoring (QCM-D) has recently emerged as a powerful alternative for the phenotypic detection of EVs, offering multiple modes of analyte discrimination by frequency and dissipation. While providing rich data for sensor development, further progress is required to reduce detection limits and fully exploit the technique’s potential within biosensing. Herein, we investigate the impact of nanostructuring the sensor electrode surface for enhancing its detection capabilities. We employ self-assembly of the block copolymer polystyrene-block-poly(4-vinylpyridine) to create well defined 2D gold islands via selective impregnation of the pyridine domain with gold precursors and subsequent removal of the template. When matched to the EV length scale, we find a 4-fold improvement in sensitivity despite a 4-fold reduction in area for analyte and ligand anchoring in comparison to a flat sensor surface. Creation of tailored and confined sensing regions interspersed by non-binding silica provides optimal spatial orientation for EV capture with reduced steric effects and negative cooperativity of grafted antibodies, offering a promising route for enhanced binding efficiency and performance of sensor platforms.

HIV non-progression despite persistent viraemia is rare among antiretroviral therapy (ART)-naïve adults, but relatively common among ART-naïve children. Previous studies indicate that ART-naïve paediatric slow-progressors (PSPs) adopt immune evasion strategies similar to those described in the SIV natural hosts. However, the mechanisms underlying this immunophenotype are not well understood. In a cohort of early-treated infants who underwent analytical treatment interruption (ATI) after 12 months of ART, expression of PD-1 on CD8+ T-cells immediately prior to ATI was the main predictor of slow progression during ATI (r=0.77, p=0.002). PD-1+ CD8+ T-cell frequency was also negatively correlated with CCR5 (r=-0.74, p=0.005) and HLA-DR (r=-0.63, p=0.02) expression on CD4+ T-cells and predicted stronger HIV-specific T-lymphocyte responses. In the CD8+ T-cell compartment of PSPs, we identified an enrichment of stem-like TCF-1+PD-1+ memory cells, whereas paediatric progressors and viraemic adults were populated with a terminally exhausted PD-1+CD39+ population. TCF-1+PD-1+ expression on CD8+ T-cells was associated with higher proliferative activity (r=0.41, p=0.03) and stronger Gag-specific effector functionality. These data prompt the hypothesis that the proliferative burst potential of stem-like HIV-specific cytotoxic cells could be exploited in therapeutic strategies to boost the antiviral response and facilitate remission in early-ART-treated infants with a preserved and non-exhausted T-cell compartment.

This work reports the fabrication of nanomagnetite- and nanotitania-incorporated polyacrylonitrile nanofibers (MTPANs) by an electrospinning process, which has the potential to be used as a membrane material for the selective removal of Cd(II) and As(V) in water. The fiber morphology was characterized by scanning electron microscopy (SEM). The incorporation of nanomagnetite and nanotitania in the composite fiber matrix was confirmed by energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. The fibers doped with nanomagnetite and nanotitania (MPAN and TPAN fibers, respectively), as well as MTPAN and neat polycrylonitrile (PAN) fibers, after thermally stabilizing at 275 °C in air, were assessed for their comparative As(V)- and Cd(II)-ion removal capacities. The isotherm studies indicated that the highest adsorption of Cd(II) was shown by MTPAN, following the Langmuir model with a qm of 51.5 mg/m2. On the other hand, MPAN showed the highest As(V)adsorption capacity, following the Freundlich model with a KF of 0.49. The mechanism of adsorption of both Cd(II) and As(V) by fibers was found to be electrostatically driven, which was confirmed by correlating the point of zero charges (PZC) exhibited by fibers with the pH of maximum ion adsorptions. The As(V) adsorption on MPAN occurs by an inner-sphere mechanism, whereas Cd(II) adsorption on MTPAN is via both surface complexation and an As(V)-assisted inner-sphere mechanism. Even though the presence of coexistent cations, Ca(II) and Mg(II), has been shown to affect the Cd(II) removal by MTPAN, the MTPAN structure shows >50% removal efficiency even for minute concentrations (0.5 ppm) of Cd(II) in the presence of high common ion concentrations (10 ppm). Therefore, the novel polyacrylonitrile-based nanofiber material has the potential to be used in polymeric filter materials used in water purification to remove As(V) and Cd(II) simultaneously.

Prenatal exposure to second-hand smoke (SHS) is associated with increased neurodevelopmental problems in children, however, its impact on the risk of developmental coordination disorder (DCD) in preschoolers have not been studied thoroughly. Herein, we probed this association based on a nationwide retrospective cohort study of 149,005 preschoolers in China. We divided the objects into the prenatal SHS-exposed group or the no prenatal smoke exposed group (NS-exposed group). Preschoolers were assessed for motor proficiency by the Chinese version of Little Developmental Coordination Disorder Questionnaire (LDCDQ). Multivariable logistic regression was used to evaluate the associations. The prevalence of prenatal SHS exposure was 23.89%. Generally, the prevalence of suspected DCD was significantly higher in prenatal SHS-exposed group (16.38% VS. 14.19%, P < 0.001). With the increase of age, the mean total scores of LDCDQ of both boys and girls increased gradually; and the prevalence of suspected DCD in girls was higher than that in boys in the same age group. After adjusting for covariates, prenatal SHS exposure had the negative association with the total score of LDCDQ and increased the risk of suspected DCD. Our results suggest a need for interventions designed to reduce maternal SHS exposure during pregnancy, early screen for DCD and increase targeted movement and coordination skill training for vulnerable children.

Since its discovery, small interfering RNA (siRNA) has revolutionised the field of gene silencing therapy because of its great potential in targeting diseases that were considered untreatable. Although several siRNA drugs are already in the market, many challenges remain to be addressed before siRNA therapies can realise their full potential including extra-hepatic delivery and improved endosomal escape. Dendritic polypeptides are attractive carriers for siRNA because of their wide range of defined structures that can be made. In this work, we report the on-going development of histidylated poly(lysine) dendrimers (G3KH) as carriers for siRNA. Histidine-capped lysine dendrimers were synthesised via HBTU/HOBt coupling reaction in solution. 1H NMR data confirms the structure of G3KH dendrimers; however, although traces of impurities were also observed. Histidylated lysine dendrimers could efficiently condense short double-strand DNA oligos (used as a model for siRNA) to form cationic nanoparticles of less than 100 nm in size

The overexpression of heat shock proteins (HSPs) in tumor cells can activate inherent defense mechanisms during hyperthermia-based treatments, inducing thermoresistance and thus diminishing the treatment efficacy. Here, we report a distinct “non-inhibitor involvement” strategy to address this issue through engineering a calcium-based nanocatalyst (G/A@CaCO3-PEG). The constructed nanocatalyst consists of calcium carbonate (CaCO3)-supported glucose oxidase (GOD) and 2D antimonene quantum dots (AQDs), with further surface modification by lipid bilayers and polyethylene glycol (PEG). The engineered G/A@CaCO3-PEG nanocatalyst features prolonged blood circulation, which is stable at neutral pH but rapidly degrades under mildly acidic tumor microenvironment, resulting in rapid release of drug cargo in the tumor microenvironment. The integrated GOD effectively catalyzes the depletion of glucose for reducing the supplies of adenosine triphosphate (ATP) and subsequent down-regulation of HSP expression. This effect then augments the therapeutic efficacy of photothermal hyperthermia induced by 2D AQDs upon irradiation with near-infrared light as assisted by reversing the cancer cells’ thermoresistance. Consequently, synergistic antineoplastic effects can be achieved via low-temperature photothermal therapy. Systematic in vitro and in vivo evaluations have demonstrated that G/A@CaCO3-PEG nanocatalysts feature potent antitumor activity with a high tumor-inhibition rate (83.92%). This work thus paves an effective way for augmenting the hyperthermia-based tumor treatments via restriction of the ATP supply., Graphical abstract Image 1, Highlights • A concept of “non-inhibitor involvement” strategy is proposed for reversing tumor cell thermoresistance. • The ingenious integration of calcium biomineral and single-element 2D antimonene quantum dots was firstly demonstrated. • The engineered nanocatalysts highlights the high potential of the ATP restriction and HSP inhibition in cancer treatment. • This work provides an effective strategy for augmenting hyperthermia-based treatments efficacy.

Extracellular vesicles (EVs) are routinely released from nearly all cell types as transport vehicles and for cell communication. Crucially, they contain biomolecular content for the identification of health and disease states that can be detected from readily accessible physiological fluids, including urine, plasma, or saliva. Despite their clinical utility within noninvasive diagnostic platforms such as liquid biopsies, the currently available portfolio of analytical approaches are challenged by EV heterogeneity in size and composition, as well as the complexity of native biofluids. Quartz crystal microbalance with dissipation monitoring (QCM-D) has recently emerged as a powerful alternative for the phenotypic detection of EVs, offering multiple modes of analyte discrimination by frequency and dissipation. While providing rich data for sensor development, further progress is required to reduce detection limits and fully exploit the technique’s potential within biosensing. Herein, we investigate the impact of nanostructuring the sensor electrode surface for enhancing its detection capabilities. We employ self-assembly of the block copolymer polystyrene-block-poly(4-vinylpyridine) to create well defined 2D gold islands via selective impregnation of the pyridine domain with gold precursors and subsequent removal of the template. When matched to the EV length scale, we find a 4-fold improvement in sensitivity despite a 4-fold reduction in area for analyte and ligand anchoring in comparison to a flat sensor surface. Creation of tailored and confined sensing regions interspersed by non-binding silica provides optimal spatial orientation for EV capture with reduced steric effects and negative cooperativity of grafted antibodies, offering a promising route for enhanced binding efficiency and performance of sensor platforms.

Research into extracellular vesicles (EVs) has grown significantly over the last few decades with EVs being widely regarded as a source of biomarkers for human health and disease with massive clinical potential. Secreted by every cell type in the body, EVs report on the internal cellular conditions across all tissue types. Their presence in readily accessible biofluids makes the potential of EV biosensing highly attractive as a noninvasive diagnostic platform via liquid biopsies. However, their small size (50-250 nm), inherent heterogeneity, and the complexity of the native biofluids introduce challenges for effective characterization, thus, limiting their clinical utility. This has led to a surge in the development of various novel EV biosensing techniques, with capabilities beyond those of conventional methods that have been directly transferred from cell biology. In this review, key detection principles used for EV biosensing are summarized, with a focus on some of the most recent and fundamental developments in the field over the last 5 years. This article is categorized under: Diagnostic Tools > Biosensing Diagnostic Tools > In Vitro Nanoparticle-Based Sensing.