Microfluidic System Consisting of a Magnetic 3D-Printed Microchannel Filter for Isolation and Enrichment of Circulating Tumor Cells Targeted by Anti-HER2/MOF@Ferrite Core-Shell Nanostructures: A Theranostic CTC Dialysis System.

Low number of circulating tumor cells (CTCs) in the blood samples and time-consuming properties of the current CTC isolation methods for processing a small volume of blood are the biggest obstacles to CTC usage in practice. Therefore, we aimed to design a CTC dialysis system with the ability to process cancer patients' whole blood within a reasonable time. Two strategies were employed for developing this dialysis setup, including (i) synthesizing novel in situ core-shell Cu ferrites consisting of the Cu-CuFe2O4 core and the MIL-88A shell, which are targeted by the anti-HER2 antibody for the efficient targeting and trapping of CTCs; and (ii) fabricating a microfluidic system containing a three-dimensional (3D)-printed microchannel filter composed of a polycaprolactone/Fe3O4 nanoparticle composite with pore diameter less than 200 µm on which a high-voltage magnetic field is focused to enrich and isolate the magnetic nanoparticle-targeted CTCs from a large volume of blood. The system was assessed in different aspects including capturing the efficacy of the magnetic nanoparticles, CTC enrichment and isolation from large volumes of human blood, side effects on blood cells, and the viability of CTCs after isolation for further analysis. Under the optimized conditions, the CTC dialysis system exhibited more than 80% efficacy in the isolation of CTCs from blood samples. The isolated CTCs were viable and were able to proliferate. Moreover, the CTC dialysis system was safe and did not cause side effects on normal blood cells. Taken together, the designed CTC dialysis system can process a high volume of blood for efficient dual diagnostic and therapeutic purposes.

Challenges and Opportunities of Using Fluorescent Metal Nanocluster-Based Colorimetric Assays in Medicine.

Development of rapid colorimetric methods based on novel optical-active metal nanomaterials has provided methods for the detection of ions, biomarkers, cancers, etc. Fluorescent metal nanoclusters (FMNCs) have gained a lot of attention due to their unique physical, chemical, and optical properties providing numerous applications from rapid and sensitive detection to cellular imaging. However, because of very small color changes, their colorimetric applications for developing rapid tests based on the naked eye or simple UV-vis absorption spectrophotometry are still limited. FMNCs with peroxidase-like activity have significant potential in a wide variety of applications, especially for point-of-care diagnostics. In this review, the effect of using various capping agents and metals for the preparation of nanoclusters in their colorimetric sensing properties is explored, and the synthesis and detection mechanisms and the recent advances in their application for ultrasensitive chemical and biological analysis regarding human health are highlighted. Finally, the challenges that remain as well as the future perspectives are briefly discussed. Overcoming these limitations will allow us to expand the nanocluster's application for colorimetric diagnostic purposes in medical practice.

Synthesis of Fe-Doped Peroxidase Mimetic Nanozymes from Natural Hemoglobin for Colorimetric Biosensing and In Vitro Anticancer Effects.

Despite their efficiency and specificity, the instability of natural enzymes in harsh conditions has inspired researchers to replace them with nanomaterials. In the present study, extracted hemoglobin from blood biowastes was hydrothermally converted to catalytically active carbon nanoparticles (BDNPs). Their application as nanozymes for the colorimetric biosensing of H2O2 and glucose and selective cancer cell-killing ability was demonstrated. Particles that were prepared at 100 °C (BDNP-100) showed the highest peroxidase mimetic activity, with Michaelis-Menten constants (Km) of 11.8 mM and 0.121 mM and maximum reaction rates (Vmax) of 8.56 × 10-8 mol L-1 s-1 and 0.538 × 10-8 mol L-1 s-1, for H2O2 and TMB, respectively. The cascade catalytic reactions, catalyzed by glucose oxidase and BDNP-100, served as the basis for the sensitive and selective colorimetric glucose determination. A linear range of 50-700 µM, a response time of 4 min, a limit of detection (3σ/N) of 40 µM, and a limit of quantification (10σ/N) of 134 µM was achieved. In addition, the reactive oxygen species (ROS)-generating ability of BDNP-100 was employed for evaluating its potential in cancer therapy. Human breast cancer cells (MCF-7), in the forms of monolayer cell cultures and 3D spheroids, were studied by MTT, apoptosis, and ROS assays. The in vitro cellular experiments showed dose-dependent cytotoxicity of BDNP-100 toward MCF-7 cells in the presence of 50 µM of exogenous H2O2. However, no obvious damage was induced to normal cells in the same experimental conditions, verifying the selective cancer cell-killing ability of BDNP-100.

Immunomagnetic Isolation of HER2-Positive Breast Cancer Cells Using a Microfluidic Device.

Analysis of circulating tumor cells (CTCs) as a tool for monitoring metastatic cancers, early diagnosis, and evaluation of disease prognosis paves the way toward personalized cancer treatment. Developing an effective, feasible, and low-cost method to facilitate CTC isolation is, therefore, vital. In the present study, we integrated magnetic nanoparticles (MNPs) with microfluidics and used them for the isolation of HER2-positive breast cancer cells. Iron oxide MNPs were synthesized and functionalized with the anti-HER2 antibody. The chemical conjugation was verified by Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, and dynamic light scattering/zeta potential analysis. The specificity of the functionalized NPs for the separation of HER2-positive from HER2-negative cells was demonstrated in an off-chip test setting. The off-chip isolation efficiency was 59.38%. The efficiency of SK-BR-3 cell isolation using a microfluidic chip with a S-shaped microchannel was considerably enhanced to 96% (a flow rate of 0.5 mL/h) without chip clogging. Besides, the analysis time for the on-chip cell separation was 50% faster. The clear advantages of the present microfluidic system offer a competitive solution in clinical applications.

A micropillar array-based microfluidic chip for label-free separation of circulating tumor cells: The best micropillar geometry?

INTRODUCTION: The information derived from the number and characteristics of circulating tumor cells (CTCs), is crucial to ensure appropriate cancer treatment monitoring. Currently, diverse microfluidic platforms have been developed for isolating CTCs from blood, but it remains a challenge to develop a low-cost, practical, and efficient strategy. OBJECTIVES: This study aimed to isolate CTCs from the blood of cancer patients via introducing a new and efficient micropillar array-based microfluidic chip (MPA-Chip), as well as providing prognostic information and monitoring the treatment efficacy in cancer patients. METHODS: We fabricated a microfluidic chip (MPA-Chip) containing arrays of micropillars with different geometries (lozenge, rectangle, circle, and triangle). We conducted numerical simulations to compare velocity and pressure profiles inside the micropillar arrays. Also, we experimentally evaluated the capture efficiency and purity of the geometries using breast and prostate cancer cell lines as well as a blood sample. Moreover, the device's performance was validated on 12 patients with breast cancer (BC) in different states. RESULTS: The lozenge geometry was selected as the most effective and optimized micropillar design for CTCs isolation, providing high capture efficiency (>85 %), purity (>90 %), and viability (97 %). Furthermore, the lozenge MPA-chip was successfully validated by the detection of CTCs from 12 breast cancer (BC) patients, with non-metastatic (median number of 6 CTCs) and metastatic (median number of 25 CTCs) diseases, showing different prognoses. Also, increasing the chemotherapy period resulted in a decrease in the number of captured CTCs from 23 to 7 for the metastatic patient. The MPA-Chip size was only 0.25 cm2 and the throughput of a single chip was 0.5 ml/h, which can be increased by multiple MPA-Chips in parallel. CONCLUSION: The lozenge MPA-Chip presented a novel micropillar geometry for on-chip CTC isolation, detection, and staining, and in the future, the possibilities can be extended to the culture of the CTCs.

Microfluidic-assisted synthesis and modeling of stimuli-responsive monodispersed chitosan microgels for drug delivery applications.

Droplet microfluidic has been established to synthesize and functionalize micro/nanoparticles for drug delivery and screening, biosensing, cell/tissue engineering, lab-on-a-chip, and organ-on-a-chip have attracted much attention in chemical and biomedical engineering. Chitosan (CS) has been suggested for different biomedical applications due to its unique characteristics, such as antibacterial bioactivities, immune-enhancing influences, and anticancer bioactivities. The simulation results exhibited an alternative for attaining visions in this complex method. In this regard, the role of the flow rate ratio on the CS droplet features, including the generation rate and droplet size, were thoroughly described. Based on the results, an appropriate protocol was advanced for controlling the CS droplet properties for comparing their properties, such as the rate and size of the CS droplets in the microchip. Also, a level set (LS) laminar two-phase flow system was utilized to study the CS droplet-breaking process in the Flow Focused-based microchip. The outcomes demonstrated that different sizes and geometries of CS droplets could be established via varying the several parameters that validated addressing the different challenges for several purposes like drug delivery (the droplets with smaller sizes), tissue engineering, and cell encapsulation (the droplets with larger sizes), lab-on-a-chip, organ-on-a-chip, biosensing and bioimaging (the droplets with different sizes). An experimental study was added to confirm the simulation results. A drug delivery application was established to verify the claim.