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[134] ACS Applied Materials & Interfaces, 2024

Exosome Precipitation by Ionic Strength Modulation: ExoPRISM

Vijaya Sunkara, Juhee Park, Jiyun Han, Jonathan Sabaté Del Río, Hyun-Ju Cho, In-Jae Oh, Yoon-Kyoung Cho*

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.

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[133] Advanced Healthcare Materials, 2023, https://doi.org/10.1002/adhm.202302121

Platelet Membrane-Enclosed Bioorthogonal Catalysis for Combating Dental Caries

Mamata Karmacharya, Sumit Kumar,* Yoon Jeong Choi, Yoon-Kyoung Cho*

"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.

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[132] Neoplasia, 42, 100906, 2023

Nuclear morphology predicts cell survival to cisplatin chemotherapy

Chi-Ju Kim*, Anna LK Gonye, Kevin Truskowski, Cheng-Fan Lee, Yoon-Kyoung Cho, Robert H Austin, Kenneth J Pienta, Sarah R Amend*

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.

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[131] Front. Immunol., 14 (2023) https://doi.org/10.3389/fimmu.2023.1129600

A machine learning approach to discover migration modes and transition dynamics of heterogeneous dendritic cells

Taegeun Song, Yongjun Choi, Jae-Hyung Jeon* and Yoon-Kyoung Cho*

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.

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[130] J. Funct. Biomater. 14(2), 117 (2023)

Tuning the Extracellular Vesicles Membrane through Fusion for Biomedical Applications

Mamata Karmacharya, Sumit Kumar*, Yoon-Kyoung Cho*

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.

Invited review
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[129] Biosensors and Bioelectronics, 224, 115057, (2023)

Integrated technologies for continuous monitoring of organs-on-chips: Current challenges and potential solutions

Jonathan Sabaté del Río, Jooyoung Ro , Heejeong Yoon, Tae-Eun Park*, Yoon-Kyoung Cho*

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.

Invited review
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[128] Small, 19, 2202962 [2023]

Bridging the Gap between Nonliving Matter and Cellular Life

Sumit Kumar,Mamata Karmacharya,Yoon-Kyoung Cho*

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.

invited review
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[127] Lab Chip, 22, 2726-274, (2022)

Prediction of tumor metastasis via extracellular vesicles-treated platelet adhesion on a blood vessel chip

Junyoung Kim, Vijaya Sunkara, Jungmin Kim, Jooyoung Ro, Chi-Ju Kim, Elizabeth Maria Clarissa, Sung Wook Jung, Hee Jin Lee, and Yoon-Kyoung Cho*

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.

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[126] Advance Materials, 34, 2200981 (2022)

SEEDING to Enable Sensitive Electrochemical Detection of Biomarkers in Undiluted Biological Samples

Jonathan Sabaté del Río,Hyun-Kyung Woo,Juhee Park,Hong Koo Ha,Jae-Ryong Kim,Yoon-Kyoung Cho*

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.

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[125] Analyst, Analyst, 147, 2023-2034 (2022)

Recent advances in spheroid-based microfluidic models to mimic tumour microenvironment

Jooyoung Ro, Junyoung Kim, Yoon-Kyoung Cho

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.

This article is part of the themed collections: Analyst Review Articles 2022 and Analyst HOT Articles 2022
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[124] Journal of Visualized Experiments : Jove. 2022 Feb(180). DOI: 10.3791/62836

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

Chi-Ju Kim§*, Morgan D. Kuczler§, Liang Dong, Junyoung Kim, Sarah R. Amend, Yoon-Kyoung Cho*, Kenneth J. Pienta

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.

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[123] Translational Oncology, 16 (2022), 101321

Clinical Significance of Circulating Tumor Cells in Unresectable Pancreatic Ductal Adenocarcinomas

Hyemin Kim, Chan Mi Heo, Jinmyeong Oh, Eun Mi Lee, Juhee Park, Se-Hoon Lee, Kwang Hyuck Lee, Kyu Taek Lee, Jong Kyun Lee, Yoon-Kyoung Cho, Joo Kyung Park

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).

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[122] Lab on a Chip, 22, 377-386 (2022)

Exhausted mature dendritic cells exhibit a slower and less persistent random motility but retain chemotaxis against CCL19

Yongjun Choi, Vijaya Sunkara, Yeojin Lee, and Yoon-Kyoung Cho*

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.

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[121] Cells, 10(12), 3362 (2021)

Dendritic Cell Migration Is Tuned by Mechanical Stiffness of the Confining Space

Yongjun Choi, Jae-Eun Kwon, and Yoon-Kyoung Cho*

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.

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[120] Lab on a Chip, 21, 4707-4715 (2021)

Fully automated light transmission aggregometry on a disc for platelet function tests

Chi-Ju Kim†, Jungmin Kim†, Jonathan Sabaté del Río, Dong Yeob Kia, Junyoung Kim, Yoon-Kyoung Cho*,

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.

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