Innovative PNA-LB mediated allele-specific LAMP for KRAS mutation profiling on a compact lab-on-a-disc device.

Tailored healthcare, an approach focused on individual patients, requires integrating emerging interdisciplinary technologies to develop accurate and user-friendly diagnostic tools. KRAS mutations, prevalent in various common cancers, are crucial determinants in selecting patients for novel KRAS inhibitor therapies. This study presents a novel state-of-the-art Lab-on-a-Disc system utilizing peptide nucleic acids-loop backward (PNA-LB) mediated allele-specific loop-mediated isothermal amplification (LAMP) for detecting the frequent G12D KRAS mutation, signifying its superiority over alternative mutation detection approaches. The designed Lab-on-a-Disc system demonstrated exceptional preclinical and technical precision, accuracy, and versatility. By applying varying cutoff values to PNA- LB LAMP reactions, the assay's sensitivity and specificity were increased by 80 % and 90 %, respectively. The device's key advantages include a robust microfluidic Lab-on-a-Disc design, precise rotary control, and a cutting-edge induction heating module. These features enable multiplexing of LAMP reactions with high reproducibility and repeatability, with CV% values less than 3.5 % and 5.5 %, respectively. The device offers several methods for accurate endpoint result detection, including naked-eye observation, RGB image analysis using Python code, and time of fluorescence (Tf) values. Preclinical specificity and sensitivity, assessed using different cutoffs for Eva-Green fluorescence Tf values and pH-sensitive dyes, demonstrated comparable performance to the best standard methods. Overall, this study represents a significant step towards tailoring treatment strategies for cancer patients through precise and efficient mutation detection technologies.

In vitro and in vivo investigation of chitosan/silk fibroin injectable interpenetrating network hydrogel with microspheres for cartilage regeneration.

Articular cartilage is an avascular and almost acellular tissue with limited self-regenerating capabilities. Although injectable hydrogels have garnered a lot of attention as a promising treatment, a biocompatible hydrogel with adequate mechanical properties is yet to be created. In this study, an interpenetrating network hydrogel comprised of chitosan and silk fibroin was created through electrostatic and hydrophobic bonds, respectively. The polymeric network of the scaffold combined an effective microenvironment for cell activity with enhanced mechanical properties to address the current issues in cartilage scaffolds. Furthermore, microspheres (MS) were utilized for a controlled release of methylprednisolone acetate (MPA), around ~75 % after 35 days. The proposed scaffolds demonstrated great mechanical stability with ~0.047 MPa compressive moduli and ~145 kPa compressive strength. Moreover, the degradation rate of the samples (~45 % after 35 days) was optimized to match neo-cartilage formation. Furthermore, the use of natural biomaterials yielded good biocompatibility with ~76 % chondrocyte viability after 7 days. According to gross observation after 12 weeks the defect site of the treated groups was filled with minimally discernible boundary. These results were confirmed by histopathology assays were the treated groups showed higher chondrocyte count and collagen type II expression.

Fabrication of gelatin-based antibacterial bilayer wound dressing using direct writing and electrospinning methods.

Fabricating a fibrous well-ordered wound dressing for accelerating full-thickness wounds is a desirable treatment vector. Here, through modifications in the material extrusion device and adding a pneumatic-based injection, a material extrusion method for gelatin was introduced with the ability to fabricate 3D structure with repeat layers to support cell activity for the under layer. Furthermore, in the upper layer, the co-electrospinning of PU with gelatin was designed to simultaneously exploit the oxygen permeability and mechanical stability of PU with regenerative properties and collagen-like structure of gelatin. Moreover, zinc oxide nanoparticles (ZnO) was added into the 3D-printed under layer to synergistically benefit from the antibacterial properties of ZnO and the excellent biocompatibility of gelatin. The controllable porosity of the under layer, enabled through the additive manufacturing method, was adjusted to mimic the extracellular matrix of natural tissue with around (127.28 ±â€¯20.70) µm pore size after swelling with smooth fibers. S. aureus, E. coli, Bacillus subtilis, and Pseudomonas with inhibition zone diameters at ∼ 2.14 cm and ∼ 1.96 cm, ∼ 4.01 cm, and ∼ 2.24 cm, respectively. Moreover, the scaffold showed great biocompatibility toward fibroblast cells after 7 days of cell culture with ∼ 89 % cell viability.

Simple Method for On-Demand Droplet Trapping in a Microfluidic Device Based on the Concept of Hydrodynamic Resistance.

We demonstrate an innovative method to catch the desired droplets from a train of droplets and immobilize them in traps located in an integrated microfluidic device. To this end, water-in-oil droplets are generated in a flow-focusing junction and then guided to a channel connected to chambers designated for on-demand droplet trapping. Each chamber is connected to a side channel through a batch of microposts. The side channels are also connected to the flexible poly(vinyl chloride) tubes, which can be closed by attaching binder clips. The hydrodynamic resistance of the routes in the device can be changed by opening and closing the binder clips. In this way, droplets are easily guided into individual traps based on the user's demand. A set of numerical simulations was also conducted to investigate the authenticity of the employed idea and to find the optimal geometry for implementing our strategy. This simple method can be easily employed for on-demand droplet trapping without using on-chip valves or complex off-chip actuators proposed in previous studies.

Microfluidic sperm sorting selects a subpopulation of high-quality sperm with a higher potential for fertilization.

STUDY QUESTION: Is a microfluidic sperm sorter (MSS) able to select higher quality sperm compared to conventional methods? SUMMARY ANSWER: The MSS selects sperm with improved parameters, lower DNA fragmentation, and higher fertilizing potential. WHAT IS KNOWN ALREADY: To date, the few studies that have compared microfluidics sperm selection with conventional methods have used heterogeneous study population and have lacked molecular investigations. STUDY DESIGN, SIZE, DURATION: The efficiency of a newly designed MSS in isolating high-quality sperm was compared to the density-gradient centrifugation (DGC) and swim-up (SU) methods, using 100 semen samples in two groups, during 2023-2024. PARTICIPANTS/MATERIALS, SETTING, METHODS: Semen specimens from 50 normozoospermic and 50 non-normozoospermic men were sorted using MSS, DGC, and SU methods to compare parameters related to the quality and fertilizing potential of sperm. The fertilizing potential of sperm was determined by measurement of phospholipase C zeta (PLCζ) and post-acrosomal sheath WW domain-binding protein (PAWP) expression using flow cytometry, and the chromatin dispersion test was used to assess sperm DNA damage. MAIN RESULTS AND THE ROLE OF CHANCE: In both normozoospermic and non-normozoospermic groups, the MSS-selected sperm with the highest progressive motility, PLCζ positive expression and PLCζ and PAWP fluorescence intensity the lowest non-progressive motility, and minimal DNA fragmentation, compared to sperm selected by DGC and SU methods (P < 0.05). LIMITATION, REASONS FOR CAUTION: The major limitations of our study were the low yield of sperm in the MSS chips and intentional exclusion of severe male factor infertility to yield a sufficient sperm count for molecular experiments; thus testing with severe oligozoospermic semen and samples with low count and motility is still required. In addition, due to ethical considerations, at present, it was impossible to use the sperm achieved from MSS in the clinic to assess the fertilization rate and further outcomes. WIDER IMPLICATIONS OF THE FINDINGS: Our research presents new evidence that microfluidic sperm sorting may result in the selection of high-quality sperm from raw semen. This novel technology might be a key to improving clinical outcomes of assisted reproduction in infertile patients. STUDY FUNDING/COMPETING INTEREST(S): The study is funded by the Iran University of Medical Sciences and no competing interest exists. TRIAL REGISTRATION NUMBER: N/A.

Design and evaluation of loop-mediated isothermal amplification for rapid detection of Enterocytozoon bieneusi.

Enterocytozoon bieneusi is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly E. bieneusi, are frequently reported from waterborne and foodborne outbreaks. Therefore, early detection is crucial in clinics and outbreak investigations. This study aimed to design a loop-mediated isothermal amplification (LAMP) for rapid detection of E. bieneusi. Total DNA was extracted from 30 E. bieneusi -positive samples, which had been confirmed with nested PCR. LAMP primers were designed based on the identical fragment of small subunit ribosomal RNA (SSU rRNA) gene. LAMP reactions were performed at 63 °C for 60 min. The sensitivity and specificity of the assay were analyzed and the results of amplification were compared to real-time PCR. Our results showed that the LAMP assay successfully amplified 25/30 (83.3%) samples. The specificity results indicated no false positive with other microorganisms. Furthermore, the LAMP method exhibited a sensitivity (limit of detection, LoD) as low as 34 ag/µL of total DNA. Compared to the LAMP assay, real-time PCR was able to detect all 30 nested PCR-positive samples. Our findings showed that the LAMP assay was able to detect 83.3% of E. bieneusi-positive samples. Although the current assay was not able to detect all nested PCR-positive samples, the lack of need for specific instruments, rapid processes, and high specificity makes LAMP assay a suitable tool for screening.

In-silico study of drug delivery to atherosclerosis in the human carotid artery using metal-organic frameworks based on adhesion of nanocarriers.

This study investigates nanocarriers (NCs) for drug delivery targeting carotid artery atherosclerosis. This targeted drug delivery mechanism is based on ligand-receptor bindings facilitated by coating NCs with P-selectin aptamers, which exhibit high affinities for P-selectin plaque receptors. Recognizing the significant advantages of metal-organic frameworks (MOFs), such as their high drug-loading percentages, we chose them as nanocarriers for this research. Our evaluation considers critical factors: NC surface density (the number of attached nanocarriers per unit of plaque area), toxicity (percentage of NCs missing the target), and efficient drug transfer to plaque tissue. Employing molecular dynamics (MD) for drug loading calculations via van der Waals interactions and computational fluid dynamics (CFD) for toxicity, surface density, and drug transfer assessments, we achieve a comprehensive analysis. A cardiac cycle-based metric guides optimal MOF release conditions, establishing an ideal dosage of 600 NCs per cycle. MOF-801 exhibits outstanding drug delivery performance, particularly in plaque targeting. While a magnetic field enhances NC adhesion, its impact on drug transfer is limited, emphasizing the need for further optimization in magnetic targeting for NC-based therapies. This study provides crucial insights into NC drug delivery performance in carotid artery atherosclerosis, advancing the field of targeted drug delivery for atherosclerosis treatment.

Fabrication and characterization of an antibacterial chitosan-coated allantoin-loaded NaCMC/SA skin scaffold for wound healing applications.

The field of tissue engineering has recently emerged as one of the most promising approaches to address the limitations of conventional tissue replacements for severe injuries. This study introduces a chitosan-coated porous skin scaffold based on sodium carboxymethyl cellulose (NaCMC) and sodium alginate (SA) hydrogels, incorporating allantoin (AL) as an antibacterial agent. The NaCMC/SA hydrogel was cross-linked with epichlorohydrin (ECH) and freeze-dried to obtain a three-dimensional porous structure. The coated and non-coated scaffolds underwent comprehensive evaluation and characterization through various in-vitro analyses, including SEM imaging, swelling, degradation, and mechanical assessments. Furthermore, the scaffolds were studied regarding their allantoin (AL) release profiles, antibacterial properties, cell viability, and cell adhesion. The in-vitro analyses revealed that adding a chitosan (CS) coating and allantoin (AL) to the NaCMC/SA hydrogel significantly improved the scaffolds' antibacterial properties and cell viability. It was observed that the NaCMC:SA ratio and ECH concentration influenced the swelling capacity, biodegradation, drug release profile, and mechanical properties of the scaffolds. Samples with higher NaCMC content exhibited enhanced swelling capacity, more controlled allantoin (AL) release, and improved mechanical strength. Furthermore, the in-vivo results demonstrated that the proposed skin scaffold exhibited satisfactory biocompatibility and supported cell viability during wound healing in Wistar rats, highlighting its potential for clinical applications.

The effect of conductive aligned fibers in an injectable hydrogel on nerve tissue regeneration.

Injectable hydrogels are a promising treatment option for nervous system injuries due to the difficulty to replace lost cells and nervous factors but research on injectable conductive hydrogels is limited and these scaffolds have poor electromechanical properties. This study developed a chitosan/beta-glycerophosphate/salt hydrogel and added conductive aligned nanofibers (polycaprolactone/gelatin/single-wall carbon nanotube (SWCNT)) for the first time and inspired by natural nerve tissue to improve their biochemical and biophysical properties. The results showed that the degradation rate of hydrogels is proportional to the regrowth of axons and these hydrogels' mechanical (hydrogels without nanofibers or SWCNTs and hydrogels containing these additions have the same Young's modulus as the brain and spinal cord or peripheral nerves, respectively) and electrical properties, and the interconnective structure of the scaffolds have the ability to support cells.

A novel multifunctional chitosan-gelatin/carboxymethyl cellulose-alginate bilayer hydrogel containing human placenta extract for accelerating full-thickness wound healing.

The replication of skin's dermal and epidermal morphology within a full-thickness wound using a bi-layer hydrogel to cater to their distinct needs is a compelling pursuit. Moreover, human placenta extract (HPE), containing a diverse array of bioactive agents, has proven to be effective in promoting the wound healing process and enhancing epidermal keratinocytes. This study presents a multifunctional bi-layer hydrogel incorporating HPE for accelerating full-thickness wound healing through sustained HPE release, inhibition of bacteria invasion, and promotion of cell proliferation. The upper layer of the scaffold, known as the dressing layer, is composed of carboxymethyl cellulose and sodium alginate, serving as a supportive layer for cell proliferation. The under layer, referred to as the regenerative layer, is composed of chitosan and gelatin, providing an extracellular matrix-like, porous, moist, and antibacterial environment for cell growth. The scaffold was optimized to replicate the morphology of the dermal and epidermal layers, with suitable fibroblast infiltration and a pore size of approximately 283µm. Furthermore, the degradation rate of the samples matched the wound healing rate and persisted throughout this period. The sustained HPE release rate, facilitated by the degradation rate, was optimized to reach ~98% after 28 days, covering the entire healing period. The samples demonstrated robust antibacterial capabilities, with bacterial inhibition zone diameters of and 2.63±0.12cm for S. aureus and E. coli, respectively. The biocompatibility of the samples remained at approximately 68.33±4.5% after 21 days of fibroblast cell culture. The in vivo experiment indicated that the HPE@Bilayer hydrogel promotes the formation of new blood vessels and fibroblasts during the early stages of healing, leading to the appropriate formation of granulation tissue and a wound contraction rate of (79.31±3.1)%. Additionally, it resulted in the formation of a thick epidermal layer (keratinization) that effectively covered all the impaired areas, achieving a wound contraction rate of 95.83±6.3% at the late stage of wound healing. Furthermore, immunohistochemistry staining for CD31 and TGF-ß revealed that the HPE@Bilayer group had 22 blood vessels/field and 34%-66% immunoactive cells, respectively, after 14 days of healing. However, by day 21, angiogenesis and TGF-ß expression had declined, demonstrating that the wounds had been successfully treated with minimal scarring.

Fabrication of heparinized bi-layered vascular graft with PCL/PU/gelatin co-electrospun and chitosan/silk fibroin/gelatin freeze-dried hydrogel for improved endothelialization and enhanced mechanical properties.

Fabricating a biocompatible small-diameter vascular graft (< 6 mm) with mechanical properties similar to the natural vein and adding good anti-thrombogenic, endothelialization, and hyperplasia properties remains a challenge. To this end, we fabricated a heparinized bilayer graft to address this problem. The proposed bilayer sample consisted of a heparinized polycaprolactone (PCL), polyurethane (PU), and gelatin (G) co-electrospun inner layer and chitosan, gelatin, and silk fibroin freeze-dried hydrogel crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) outer layer. The samples exhibited great ultimate stress, Young's module, and suture retention of 4.16±0.25MPa, 8.24±2.59MPa and 4.83±0.31N, respectively. The heparin release assay indicated a sustained release profile of around 70% after 4weeks, which can be attributed to the excellent control via emulsion. Furthermore, the heparinized samples demonstrated good anti-thrombogenic properties investigated in the platelet adhesion assay. For the outer layer, the hydrogel crosslinked with non-toxic materials was prepared through the freeze-drying method to achieve high porosity (64.63%), suitable for smooth muscle cell activity. Moreover, inner and outer layers showed high cell viability toward endothelial (78.96%) and smooth muscle cells (57.77%), respectively. Overall, the proposed heparinized graft exhibited excellent potential for vascular graft regeneration.

An experimental study of centrifugal microfluidic platforms for magnetic-inertial separation of circulating tumor cells using contraction-expansion and zigzag arrays.

Cancer diagnosis has recently been at the forefront of recent medical research, with ongoing efforts to develop devices and technologies for detecting cancer in patients. One promising approach for cancer diagnosis is the detection of Circulating Tumor Cells (CTCs) in blood samples. Separating these rare cells from the diverse background of blood cells and analyzing them can provide valuable insights into the disease's stage and lethality. Here we present the design and fabrication of a centrifugal microfluidic platform on a polymeric disk that utilizes centrifugal forces for cell isolation. The separation units exploit both active and passive methods. In other words, in addition to introducing novel geometry for channels, an external magnetic field is also employed to separate the target cells from the background cells. In order for the external field to function, the CTCs must first be labeled with antibody-conjugated nanoparticles; the separation process should be then performed. Before the experimental tests, a numerical study was done to determine the optimum parameters; the angular velocity and magnetization investigations showed that 2000 rpm and 868,000 (kA/m) are the optimum conditions for the designed device to reach the efficiency of 100% for both White Blood Cells (WBCs) and CTCs. These results indicate that the passive region of the channels primarily contributes to the focusing of the target cells, and showed that the focusing effect is more pronounced in the expansion-contraction geometry compared to the zigzag geometry. Additionally, the results proved that curved channel geometries performed better than straight ones in terms of separation efficiency. However, if the separation relies solely on channel geometry, the majority of cells would be directed towards the non-target chamber, leading to suboptimal results. This is due to the direction of the forces acting on the cells. However, including an external magnetic field improves the direction of the net force and enhances the separation efficiency. Finally, the numerical and experimental results of the study were compared, and the curved expansion-contraction channel is introduced as the best geometry having 100% and ∼92% CTC separation efficiency, respectively.

A curved expansion-contraction microfluidic structure for inertial based separation of circulating tumor cells from blood samples.

The rare presence of circulating tumor cells (CTCs) in the bloodstream has made their recording and separation one of the major challenges in the recent decade. Inertia-based microfluidic systems have received more attention in CTCs separation due to their feasibility and low cost. In this research, an inertial microfluidic system is proposed using a curved expansion-contraction array (CEA) microchannel to separate CTCs from white blood cells (WBCs). First, the optimal flow rate of the proposed microfluidic device was determined to maximize the separation efficiency of the target cells (CTCs) from the non-target ones (WBCs). Then, the efficiency and purity of the straight and curved-CEA microchannels were assessed. The experimental results indiated that the proposed system (curved-CEA microchannel) can offer the highest efficiency (-80.31%) and purity (-91.32%) at the flow rate of -7.5 ml/min, exhibiting ∼11.48% increment in the efficiency compared to its straight peer.

Numerical study of a double-stair-shaped dielectrophoresis channel for continuous on-chip cell separation and lysis using finite element method.

Prognostication of numerous chronic diseases are in need of identifying circulating tumor cells (CTCs), afterwards, separating and reviving contaminated samples are required. Conventional methods of separating blood cells, namely cytometry or magnetically activated cell sorting, in many cases lose their functionality, or efficiency under different conditions. Hence microfluidic methods of separation have been implemented. Herein, an innovative integrated double stair-shaped microchannel is designed and optimized, capable of 'separation', and 'chemical lysis' simultaneously in which the lysis reagent concentration can be controlled to tune the lysis intensity. The method of insulator-based dielectrophoresis (iDEP), which is the main physics in this device, is utilized yielding maximum separation. Pivotal features of the applied voltage, the voltage difference, the angles and the number of stairs, and the width of the throat in the microchannel have been numerically explored in order to optimize the channel in terms of separation and the lysis buffer concentration. The overall state of optimum case for the voltage difference (ΔV) of 10 owns the following features: the number of stairs is 2, the angle of stairs is 110°, the width of throat is 140 µm, and the inlet voltages are 30 V and 40 V. Also, the overall state of optimum cases for delta possess the following features: the number of stairs is 2, the angle of stairs is 110°, the width of throat is 140 µm, and the inlet voltages are 30 V and 35 V.

Novel bilayer coating on gentamicin-loaded titanium nanotube for orthopedic implants applications.

Fabricating a multifunctional orthopedic implant which prevents post-surgery infection is highly desirable in advanced materials applications. However, designing an antimicrobial implant, which simultaneously promotes a sustained drug release and satisfactory cell proliferation, remains a challenge. The current study presents a drug-loaded surface-modified titanium nanotube (TNT) implant with different surface chemistry which was developed to investigate the effect of surface coating on drug release, antimicrobial activity, and cell proliferation. Accordingly, sodium alginate and chitosan were coated on the surface of TNT implants with different coating orders through layer-by-layer assembly. The coatings' swelling ratio and degradation rate were around 613% and 75%, respectively. The drug release results showed that surface-coatings prolonged the releasing profile for about 4 weeks. Chitosan coated TNTs demonstrated greater inhibition zone at 16.33mm compared with the other samples where no inhibition zone was observed. However, chitosan and alginate coated TNTs exhibited smaller inhibition zones at 48.56mm and 43.28mm, respectively, compared to bare TNT, which can be attributed to the coatings preventing the antibiotic burst release. Higher viability of cultured osteoblast cells was observed for chitosan-coated TNT as the top layer compared to the bare TNT at 12.18%, indicating improved bioactivity of TNT implants when the chitosan has the most contact with cells. Coupled with the cell viability assay, molecular dynamics (MD) simulations were carried out by placing collagen and fibronectin near the considered substrates. In agreement with cell viability results, MD simulations also indicated that chitosan had the highest adsorption energy approximately 60Kcal/mol. In summary, the proposed bilayer chitosan-coated drug-loaded TNT implant with chitosan and sodium alginate coating as the top and the bottom layers, respectively, can be a potential candidate for orthopedic applications in the light of its bacterial biofilm prevention, better osteoconductivity, and providing an adequate drug release profile.

Investigating the correlation between the protein adhesion simulation and the biocompatibility of polymeric substrate for skin-tissue-engineering applications.

Investigating the protein adhesion properties of polymeric scaffolds through computational simulations can predict the biocompatibility of scaffolds before an experimental assay is carried out. This prediction can be highly beneficial since it can cut costs and the time it takes for experimental assays. The current study aims to test the hypothesis that there is a correlation between the biocompatibility of a composite scaffold and the molecular dynamics simulations of protein adhesion. To this end, chitosan and gelatin were selected for fabricating a composite skin-tissue wound scaffold with five different polymer ratios. This polymeric blend has not been simulated for protein adhesion. The cell proliferation and viability of the samples were quantified via MTT assay using fibroblast cells. Then a series of molecular dynamics simulations were performed to measure the adhesion energy of two prominent extracellular matrix proteins - fibronectin, and collagen type I. Besides, a higher gelatin percentage in the scaffold leads to a decrease in the porosity. The results demonstrated a strong correlation between the experimental data and molecular dynamics simulations. The sample with equal amounts of chitosan and gelatin had the highest cell viability and the strongest adhesion energy, of 239 kcal mol-1 for collagen type I, and 149 kcal mol-1 for fibronectin. This correlation was also evident in other samples: samples with gelatin-to-chitosan ratios of 3 : 1 and 1 : 3 had the lowest cell viability and the weakest adhesion energy, respectively.

Metal-organic frameworks: A promising option for the diagnosis and treatment of Alzheimer's disease.

Beta-amyloid (Aß) peptide is one of the main characteristic biomarkers of Alzheimer's disease (AD). Previous clinical investigations have proposed that unusual concentrations of this biomarker in cerebrospinal fluid, blood, and brain tissue are closely associated with the AD progression. Therefore, the critical point of early diagnosis, prevention, and treatment of AD is to monitor the levels of Aß. In view of the potential of metal-organic frameworks (MOFs) for diagnosing and treating the AD, much attention has been focused in recent years. This review discusses the latest advances in the applications of MOFs for the early diagnosis of AD via fluorescence and electrochemiluminescence (ECL) detection of AD biomarkers, fluorescence detection of the main metal ions in the brain (Zn2+, Cu2+, Mn2+, Fe3+, and Al3+) in addition to magnetic resonance imaging (MRI) of the Aß plaques. The current challenges and future strategies for translating the in vitro applications of MOFs into in vivo diagnosis of the AD are discussed.

Novel size-based design of spiral microfluidic devices with elliptic configurations and trapezoidal cross-section for ultra-fast isolation of circulating tumor cells.

Investigation and analysis of circulating tumor cells (CTCs) have been valuable resources for detecting and diagnosing cancer in its early stages. Recently, enumeration and separation of CTCs via microfluidic devices have attracted significant attention due to their low cost and easy setup. In this study, novel microfluidic devices based on size-dependent cell-sorting with a trapezoidal cross-section and elliptic spiral configurations were proposed to reach label-free, ultra-fast CTCs enrichment. Firstly, the possibility and quality of separation in the devices were evaluated via a numerical simulation. Subsequently, these devices were fabricated to investigate the effects of the altering curvature and the trapezoidal cross-section on the isolation of CTCs from the peripheral blood sample at varying flow rates ranging from 0.5 mL/min to 3.5 mL/min. The experimental results indicated that the flow rate of 2.5 mL/min provided the optimal separation efficiency in the proposed devices, which was in fine agreement with the numerical analysis results. In this experiment, the purity values of CTCs between 88% and 90% were achieved, which is an indicator of the high capability of the proposed devices for the isolation and enrichment of CTCs. This strategy is hoped to overcome the limitations of classical affinity-based CTC separation approaches in the future.

Evaluating the reliability of tumour spheroid-on-chip models for replicating intratumoural drug delivery: considering the role of microfluidic parameters.

Several tumour spheroid-on-chip models have already been proposed in the literature to conduct high throughput drug screening assays. The microfluidic configurations in these models generally depend on the strategies adopted for spheroid formation and entrapment. However, it is not clear how successful they are to mimic in vivo transport mechanisms. In this study, drug transport in different tumour spheroid-on-chip models is numerically investigated under static and dynamic conditions using porous media theory. Moreover, the treatment of a solid tumour at the initial stage of development is modelled using bolus injection and continuous infusion methods. Then, the results of tumour spheroid-on-chip, including drug concentration, cell viability, as well as pressure and fluid shear stress distributions, are compared with those of the solid tumour, assuming identical transport properties in all models. Finally, a new configuration of the microfluidic device along with the optimal drug concentrations is proposed, which can well imitate a given in vivo situation.