Journal: bioRxiv

Article Title: Cell-nanoplastics association impacts cell proliferation and motility

doi: 10.64898/2026.04.03.716369

Figure Lengend Snippet: (A) Representative confocal images of MDCK II monolayers expressing CaaX-GFP to label cell membrane (green), stained with Hoechst 33342 to label cell nucleus (cyan), and pre-incubated with 20 µg/mL red fluorescent NPs (red) for 24 hours. Panel on the right is a z -stack image. (B) Cell-associated NP fluorescence of different cell lines quantified using flow cytometry. The median fluorescence intensity (MFI) of N = 3 biological replicates is plotted in the log scale. (C) A method to quantify the mass of NPs using epifluorescence microscopy and microfluidic devices. 20 to 1600 µg/mL of NPs were injected into a microfluidic channel with known height similar to tissue cells, and imaged using an epifluorescence microscope. The total fluorescence per unit area is a linear function of the NP density. n = 3 technical repeats per condition. (D) Quantification of cell-associated NP density at different NP loading times in the MDCK II monolayer. 20 µg/mL NPs were loaded for 1, 6, and 24 hours, followed by washing and epifluorescence imaging. The mass density was then calculated based on the cell height measured in (A) and the calibration curve in (C). N = 4. (E) The relative NP release at different times after 24 hours of NP loading in the MDCK II monolayer. The release fraction is the loss of fluorescence intensity compared to the intensity measured immediately after NP loading by tracking the same region of cells. N = 4. (F) Schematic of the in vivo model. C57BL/6 mice were administered 100 µL of NPs (2 mg/mL) via intravenous (i.v.) injection, then sacrificed at 1, 7, and 30 days post-injection. NP retention was assessed by IVIS imaging. (G) NP distribution in major organs and brain 30 days post-injection. (H) Representative images and total radiant efficiency of NPs in liver at 1, 7, and 30 days post-injection. (B, C, D, E, H) Error bars represent standard deviation. Student’s t -test or one-way ANOVA was used. Representative statistical significance is shown: ns for p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (A) Scale bar = 10 µm.

Article Snippet: Microfluidic channel masks were designed using AutoCAD and fabricated by FineLineImaging.

Techniques: Expressing, Membrane, Staining, Incubation, Fluorescence, Flow Cytometry, Epifluorescence Microscopy, Injection, Microscopy, Imaging, In Vivo, Standard Deviation

Journal: bioRxiv

Article Title: Cell-nanoplastics association impacts cell proliferation and motility

doi: 10.64898/2026.04.03.716369

Figure Lengend Snippet: (A) Relative cell-associated NP fluorescence in MDCK II monolayers under treatment with different inhibitors. Cells were co-incubated with 20 µg/mL NPs and inhibitors for 6 hours. NP fluorescence intensity was normalized to vehicle control (DMSO). N = 3-6. (B) Dosedependent response of NP association under EIPA (NHE1 inhibitor) treatment. N = 6. (C) Relative NP release from MDCK II monolayers under various drug treatments. All samples were first incubated with 20 µg/mL NPs for 6 hours without drug treatment, followed by extensive washing and imaging to quantify the initial level of cell-associated NPs. Cells were then incubated in NP-free media containing inhibitors for 24 hours, after which NP fluorescence was measured again. The release fraction was calculated as the relative decrease in fluorescence intensity over this period. Conditions that compromised monolayer integrity due to cytotoxicity were excluded. N = 3-6. (D and E) Dose-dependent NP release fractions under treatment with CID-1067700 (Rab7 inhibitor; D) and LY294002 (PI3K inhibitor; E). N = 3-6. (F) Quantification of trans-epithelial NP transport sing a transwell system. MDCK II monolayers grown on 1 µm pore transwells were treated with 50 µg/mL NPs and inhibitors for 24 hours. NPs in the lower compartment were quantified using microfluidic devices. Empty transwells were used as baseline controls. N = 5. (G) Quantification of trans-epithelial NP transport in the presence of Dynasore or LY294002 using the assay in (F). N = 4-5. (H) Schematic illustration of pathways affecting NP-cell association. (A-G) Error bars represent standard deviation. (A and B) Paired t -test or RM one-way ANOVA was used. (C-G) Student’s t -test or one-way ANOVA was used.

Article Snippet: Microfluidic channel masks were designed using AutoCAD and fabricated by FineLineImaging.

Techniques: Fluorescence, Incubation, Control, Imaging, Standard Deviation

Journal: Bioactive Materials

Article Title: Harnessing bi-exosome combination alleviates osteoarthritis progression

doi: 10.1016/j.bioactmat.2025.11.050

Figure Lengend Snippet: Evolution of the in vivo anti-osteoarthritic effects of Exo. (A) Anti-osteoarthritic scheme timeline: (i) Fabrication of Exo-loaded microspheres using microfluidic technology; (ii) The OA rats were indued by joint destabilization and over-erosion for 4 weeks followed by two intraarticular injections with 2 weeks interval. Measurements were conducted at the 2 weeks and 12 weeks post-2nd drug administration; (iii) Mechanical sensitivity (Von Frey) test; (iv) Mobility (Gait) analysis. Figure was created using BioRender. (B) Representative morphology of the microsphere in oil phase, scale bar: 50 μm. (C) Representative image showing DiI labeled B‐Exo and DiO labeled C‐Exo inside microsphere, scale bar: 50 μm. White dash circles indicate the microsphere outer boundaries. (D) Size distribution analysis of Exo-loaded microspheres. (E) Release curve of protein-containing Exo from Exo-loaded microspheres in vitro . (F) The pain threshold of the hind paw in rats at 12-week. (G) Quantification analysis of stride length, stand time, swing time, and swing speed of rats at 12-week. (H and J) Representative μCT images of osteophyte after 12-week joint injection, and quantification analysis of the osteophyte volume, scale bar: 5 mm. (I) The width of knee joint in rats at 12-week. (K) The changes of synovial fluid cytokine (TNF-α, IL-6, and IL-1β) levels in 12-week rat knee joints. Data are given as mean ± SD, n = 4 rats per group (except for (E), n = 3). Statistical significance was determined by one-way ANOVA and Tukey's multiple comparisons test.

Article Snippet: These two phases were mixed to form droplets at the junction within the microfluidic chip flow channel (Regenovo, DY01-1661).

Techniques: In Vivo, Labeling, In Vitro, Injection

Journal: Biosensors

Article Title: Development of Advanced Nanobiosensors and a Portable Monitoring System for Pesticide Detection at the Point of Need

doi: 10.3390/bios16020109

Figure Lengend Snippet: The key components of the microfluidic system. The microfluidic system has been designed with separate “sample” and “buffer” inlets in case sample and buffer mixing is needed.

Article Snippet: For the fabrication of the microfluidic channel, polydimethylsiloxane (PDMS) SYLGARD 184 silicone elastomer (10:1), purchased from DowCorning GmbH, was used.

Techniques:

Journal: Biosensors

Article Title: Development of Advanced Nanobiosensors and a Portable Monitoring System for Pesticide Detection at the Point of Need

doi: 10.3390/bios16020109

Figure Lengend Snippet: Calibration curve for biosensors integrated into the microfluidic chip (MFC) functionalized with ( a ) tebuconazole (TBZ)- and ( b ) lambda-cyhalothrin (CHL)-specific aptamers. ( a ) Also incorporates results of standalone sensor calibration (TBZ drop-casting) for comparison. Error bars correspond to the standard deviation after the measurement of 20 distinctive biosensors for each target analyte (i.e., TBZ and CHL), while dashed lines correspond to the linear fitting over log 10 (concentration). Selectivity of the biosensors has been studied using cross-reactive targets, i.e., difenoconazole (DFZ)/hexaconazole (HXZ) and deltamethrin (DMT)/cypermethrin (CMT) for TBZ and CHL biosensors, respectively. Inset figures show in more detail the biosensors’ response for concentrations close to their limit of detection (LoD). ( c ) Biosensor saturation data over the concentration range of 10–300 μM.

Article Snippet: For the fabrication of the microfluidic channel, polydimethylsiloxane (PDMS) SYLGARD 184 silicone elastomer (10:1), purchased from DowCorning GmbH, was used.

Techniques: Comparison, Standard Deviation, Concentration Assay

Journal: Biosensors

Article Title: Development of Advanced Nanobiosensors and a Portable Monitoring System for Pesticide Detection at the Point of Need

doi: 10.3390/bios16020109

Figure Lengend Snippet: Aptamer/NP-based biosensors integrated into a microfluidic chip for the detection of commercially available pesticides based on tebuconazole (TBZ), lambda-cyhalothrin (CHL), difenoconazole (DFZ), acetamiprid (ACTM) and deltamethrin (DMT). ( a ) Response and selectivity study for TBZ-specific biosensors for a mixture containing all five pesticides. ( b ) Response and selectivity study for CHL-specific biosensors for a mixture containing all five pesticides/fungicides. Error bars correspond to the standard deviation after the measurement of 10 distinctive biosensors for each of the three pesticides.

Article Snippet: For the fabrication of the microfluidic channel, polydimethylsiloxane (PDMS) SYLGARD 184 silicone elastomer (10:1), purchased from DowCorning GmbH, was used.

Techniques: Pesticides, Standard Deviation

Journal: Life Science Alliance

Article Title: Brain vascular stability relies on PAK2–cilia–PDGF-BB–HSPGs on basolateral side of endothelium

doi: 10.26508/lsa.202503460

Figure Lengend Snippet: (A) Human microvascular brain ECs cultured in a microfluidic chamber and stained for ciliary marker ARL13b and acetylated α-tubulin in the cross-sectional plane; the white dotted box was enlarged in the middle. (B) Transverse plane of the vessel tube. White arrowheads indicate ARL13b cilia projecting into the basolateral space. Scale bars: 100 μm. (C, C′) Fluorescent image of a 52-hpf transgenic zebrafish Tg( flk :mCherry; b-actin :Arl13b-GFP) expressing mCherry in the vasculature and mouse Arl13b-GFP fusion protein in the cilia. (C′) White boxed area is enlarged in (C′). White arrows in (C′) point to basolateral zebrafish endothelial cilium location. The scale bar in (C, C′) is 50 μm. (D) Quantification of zebrafish endothelial cilia (luminal versus abluminal location) in metencephalic artery (MTA) and middle cerebral vein (MCeV). N = 4 for each vessel group was quantified for the presence of abluminal versus luminal cilia. An unpaired t test showed no difference in the cilium location between the two vessel groups. (E) Immunohistochemistry for abluminal cilia from representative cortical microvessels from ARL13b-EGFP transgenic adult mice ( C57BL/6 background) showing luminal versus abluminal cilium localization stained with anti-GFP. The black arrow points to Arl13b-positive abluminal cilia. Scale bars: 100 μm. (E′) Quantification of Arl13b cilium length on the abluminal versus luminal side of mouse endothelial vessels. (E″) Frequency of cilia on the abluminal versus luminal side of mouse endothelial vessels. A total of 30 endothelial vessel-like structures were quantified. (F) Immunofluorescence staining of a 22-wk-old human brain for ARL13b protein (green) and CD31 vessel staining (red). White arrows point to cilia in the basolateral side of the vessel. Scale bars: 10 μm. (F′) Quantification of endothelial cilial frequency in the ventricular zone (VZ) and subventricular zone (SVZ) in human brain sections. Scale bar in (F) is 10 μm.

Article Snippet: To generate 3D EC vessels, we used the Nortis ParVivo triple-channel microfluidic chips, which we will refer to as the microphysiological systems (MPS).

Techniques: Cell Culture, Staining, Marker, Transgenic Assay, Expressing, Immunohistochemistry, Immunofluorescence