Here, we present EV precipitation by ionic strength modulation (ExoPRISM), a simple, low-cost, user-friendly, and readily adaptable approach for separating EVs in high yields without compromising their biological functions. Adding an electrolyte solution to blood plasma in small increments generates the sequential precipitation of proteins and EVs, allowing for fractional separation of EVs using low-speed centrifugation. The coprecipitated electrolytes are easily washed away, and the entire EV separation and washing process takes less than an hour. This approach successfully separates EVs from a broad range of volumes and types of biological fluids, including culture medium, urine, plasma, and serum, showing promise as a robust tool for next-generation liquid biopsies and regenerative medicine.

"bind and kill" strategy utilizing a biologically interfaced nanoreactor with platelet membrane-coated surfaces is introduced for synthesizing an antimicrobial drug through biorthogonal chemistry. The nanoreactor exploits the diverse functional proteins found in human platelets, enabling selective binding to a broad range of bacteria and effectively eliminate pathogenic biofilms formed on human teeth while maintaining excellent biocompatibility.

Chemotherapy resistance is a major factor contributing to cancer-related deaths. While researchers have studied the molecular mechanisms of resistance, less is known about the cell biology of cancer cells surviving chemotherapy. Here, we focus on prostate cancer cells treated with cisplatin and found that surviving cells grew larger, had larger nuclei, and employed efficient DNA damage repair. These cells also exhibited distinct nucleolar features and increased ribosomal RNA levels. This suggests that after chemotherapy, most cells with severe DNA damage die, while a minority adapt to a pro-survival state known as the polyaneuploid cancer cell (PACC) state. Understanding these characteristics is crucial for combating cancer resistance and recurrence.

Dendritic cells (DCs) are responsible for initiating and controlling immune responses and exhibit a heterogeneous and dynamic migratory behavior that is important for their function. We used unsupervised machine learning to analyze long-term cell migration trajectories and identified three distinct migratory modes: slow-diffusive, slow-persistent, and fast-persistent. We found that the distribution and dynamic transitions of these modes are related to the maturation status of the DCs, with immature DCs exhibiting more frequent mode changes than mature DCs. Additionally, we observed that immature DCs follow a unicyclic transition from diffusive to persistent motility, while mature DCs show no directionality in their transitions. The study highlights the complexity of biological motility and the potential of machine learning to provide new insights into this process.

EVs have a high loading capacity, bio-compatibility, and stability, making them ideal for producing effective drugs and diagnostics. The unique properties of fused EVs and the crucial design and development procedures that are necessary to realize their potential as drug carriers and diagnostic tools are examined. The promise of EVs in various stages of disease management highlights their potential role in future healthcare.

Organs-on-chips (OoCs) are biomimetic in vitro systems based on microfluidic cell cultures that recapitulate the in vivo physicochemical microenvironments and the physiologies and key functional units of specific human organs. Continuous monitoring of important quality parameters of OoCs via a label-free, non-destructive, reliable, high-throughput, and multiplex method is critical for assessing the conditions of these systems and generating relevant analytical data; moreover, elaboration of quality predictive models is required for clinical trials of OoCs. In this review, we describe recent efforts to integrate biosensing technologies into OoCs for monitoring the physiologies, functions, and physicochemical microenvironments of OoCs. Furthermore, we present potential alternative solutions to current challenges and future directions for the application of artificial intelligence in the development of OoCs and cyber-physical systems.

Though synthetic micro or nanoreactors have been used to mimic life-like functions and to learn about the fundamental biology of natural cells, constructing a life-like structure out of non-living building blocks remains a considerable challenge. In this review, we focus on hybrid approaches that use both natural and synthetic materials to mimic and interface with biological systems. Using hybrid vesicle micro or nanoreactors, bioinspired life-like functions such as chemical compartments, cascade signaling, energy generation, growth, replication, and environmental adaptation are demonstrated. Here, we discuss the strategies for constructing cell membrane-engineered hybrid synthetic vesicles and their biomedical applications, as well as the remaining challenges and future research opportunities.

Here, we found that interleukin-8 (IL-8) in cancer-derived EVs contributed to platelet activation by increasing P-selectin expression and ligand affinity, resulting in increased platelet adhesion on the human vessel-mimicking microfluidic system. Furthermore, platelet adhesion levels on vessels treated with human plasma-derived EVs demonstrated good discrimination between breast cancer patients with metastasis and those without, with the area under the curve (AUC) value of 0.88. While EpCAM expression on EVs could detect the existence of a tumor (AUC = 0.89), it performed poorly in predicting metastasis (AUC = 0.42). We believe that these findings shed light on the role of the interaction between cancer-derived EVs and platelets in pre-metastatic niche formation and tumor metastasis, potentially leading to the development of platelet–tumor interaction-based novel diagnostic and therapeutic strategies.

Electrochemical biosensors forf complex biological fluids such as plasma has been limited by the loss of sensitivity caused by biofouling. Herein, we demonstrate the preferential etching of chloride and surfactant-assisted anisotropic gold reduction to create homogeneous, nanostructured, and nanoporous gold electrodes, yielding 190 ± 20 times larger surface area within a minute without using templates. We named this process Surfactant-based Electrochemical Etch-Deposit Interplay for Nanostructure/Nanopore Growth (SEEDING). SEEDING on electrodes enhanced the sensitivity and anti-biofouling capabilities of amperometric biosensors, enabling direct analysis of tumor-derived extracellular vesicles (tEVs) in complex biofluids with a limit of detection of 300 tEVs/μL from undiluted plasma and good discrimination between patients with prostate cancer from healthy ones with an area under the curve (AUC) of 0.91 in urine and 0.90 in plasma samples.

Three-dimensional (3D) multicellular spheroid models can recapitulate the human tumour microenvironment with more accuracy than conventional cell culture models, as they include complex architectural structures and dynamic cellular interactions. This review provides an overview of the advantages of 3D spheroid cultures with a summary of the recent applications for tumour microenvironment-focused cellular interactions, as well as the studies on spheroids and external stimuli. These 3D tumour spheroid-based microfluidic devices will provide a platform for a better understanding of cellular and external interactions, as well as the discovery of cancer therapeutics.

EV uptake plays a role in intercellular communication in various research fields; cancer biology, neuroscience, and drug delivery. Fluorescently labeled EVs were prepared using a nano-filtration-based microfluidic device, Exodisc, visualized by 3D confocal microscopy, and then analyzed through advanced image-processing software. The protocol provides a robust methodology for analyzing EVs on a cellular level and a practical approach for efficient EV uptake analysis.

Here, we investigated whether longitudinal analysis of CTCs could monitor disease progression, response to chemotherapy, and survival in patients with unresectable pancreatic ductal adenocarcinoma (PDAC).

The well-coordinated migration of DCs under various immunological or inflammatory conditions is
essential to ensure an effective immune response. Here, we studied the cellular migration of exhausted DCs in tissue-mimicked confined environments. We report a unique intrinsic cell migration behaviour of exhausted mature dendritic cells, which exhibit a slower, less persistent, and less diffusive random motility, which results in the DCs remaining at the site of infection, although a well preserved CCR7-dependent chemotactic motility is maintained.

Dendritic cells (DCs), the sentinels of the immune system, are exposed to complex tissue microenvironments with a wide range of stiffnesses. Here, immature DCs were found to navigate confined spaces in a rapid and persistent manner, surveying a wide range when covered with compliant gels mimicking soft tissues. However, the speed and persistence time of random motility were both decreased by confinement in gels with higher stiffness, mimicking skin or diseased, fibrotic tissue. Our study provides evidence for a role for environmental mechanical stiffness in the surveillance strategy of immature DCs in tissues.

Platelet function tests, a group of assays that measure the ability of platelets to aggregate and promote clotting in a sample of blood, are performed in various medical fields to assess inherited platelet function disorders and monitor antiplatelet therapies. Here, we describe the design, fabrication, and operation of a centrifugal microfluidic disc that can perform a fully automated Light transmission aggregometry (LTA) assay from a small volume of a whole blood sample (<1 mL), achieving highly reproducible results (3.2% coefficient of variation) within a short period (<25 min). The assays performed with this device yield more precise and accurate results than traditional LTA because of the automation of the reaction steps, minimal human operation, and quick analysis that minimizes the adverse effects of platelet instability.

The clinical significance of circulating tumor cells (CTCs) and TWIST expression in CTCs remains unelucidated in patients with gastric cancer (GC). Here, we evaluated CTCs and TWIST expression in CTCs and explored their correlation with prognosis in patients with metastatic GC. Our study demonstrated that high levels of CTCs and TWIST (+) CTCs were associated with worse OS.

Despite recent advances, some patients with pancreatic cancer are refractory to treatment and the disease rapidly progresses, resulting in early death. The potential prognostic value of circulating tumor cells (CTCs) has been demonstrated in other cancer types, but the clinical validity in pancreatic cancer remains elusive. Here, we show that CTC clusters, which show mesenchymal characteristics and platelet marker expression, are highly correlated with poor prognosis in patients with unresectable pancreatic cancer.

Reliable, inexpensive, and rapid diagnostic tools are essential to control and prevent the spread of infectious diseases. In this Account, we detail the recent progress made by our group and others in the development of centrifugal microfluidic devices or lab-on-a-disc (LOAD) systems. Considering the urgent need for cost-effective and reliable POC infection diagnostic tools, especially in the current pandemic crisis, the current limitations and future directions of fast and broad adaptation in real-world settings are also discussed. With proper attention to key challenges and leverage with recent advances in biosensing technologies, molecular biology, nanomaterials, analytical chemistry, miniaturization, system integration, and data management, LOAD systems hold the potential to deliver POC infection diagnostic tools with unprecedented performance regarding time, accuracy, and cost. We hope the new insight and promise of LOAD systems for POC infection diagnostics presented in this Account can spark new ideas and inspire further research and development to create better healthcare systems for current and future pandemics.

The design of artificial organelles for applications in living cells faces several challenges such as cellular uptake, stability and bio compatibility. Here, fusion of exosomes creates beneficial nanoreactors and their use for compartmentalized biocatalytic cascade reactions in cells is demonstrated. Exosome membrane proteins were chemically engineered with a catechol moiety to drive fusion by supramolecular complexation to bridge the membranes. This strategy successfully encapsulated multiple enzymes and assembled the minimal electron transport chain in the plasma membrane, leading to tuneable, enhanced catalytic cascade activity capable of ATP synthesis inside of tissue spheroids. This nanoreactor was functional for many hours after uptake into living cells, showed successful penetration into tissue spheroids and repaired the damaged region by supplying ATP, all of which represent an advance in the mimicking of nature’s own organelles.

Cysteamine (CA) is a cystine depleting agent used in the treatment of cystinosis and many other diseases. However, high dose of CA can be toxic and thus point-of-care-test devices measuring blood CA level can be highly beneficial. Here, we report a highly sensitive, straightforward, and quantitative assay for the colorimetric and spectroscopic determination of CA concentration using plasmonic nanoparticles. The principle is based on the chemical etching-induced exchange of the surface ligands of plasmonic gold nanoparticles (AuNPs) upon the addition of CA. This plasmonic AuNPs sensor displays a clear red-to-blue colorimetric transition in the presence of CA among various biothiols with high specificity and sensitivity within a short time (<15 s). This fully automated lab-on-a-disc platform approach for naked eye detecting the CA concentration in human blood samples (20 μL) is highly simple and time-efficient (<6 min), and it would be potentially useful for the careful selection of CA doses in the hospital industry.