Journal: Drug Delivery and Translational Research

Article Title: Innovative microfluidic model for investigating the intestinal mucus barrier: numerical and experimental perspectives

doi: 10.1007/s13346-025-01818-8

Figure Lengend Snippet: Schematic overview of the study: Biosimilar mucus : In this study, we employed synthetic mucus to replicate human intestinal mucus. Two series of synthetic mucus were utilized to validate their suitability, with one series possessing a higher elastic modulus. In-vitro : The in-vitro section of this study involved fabricating a microfluidic chip using soft lithography. To achieve the optimal design and experimental setup, various designs were tested. Upon identifying the optimal design and experimentation method, synthetic biosimilar mucus was used to saturate the chip. Subsequently, injection of HBSS was initiated. Imaging of fluorescent particles in both the mucus and HBSS was conducted using a fluorescent microscope. Subsequently, the tracking of these particles was performed using ImageJ software to evaluate the velocity field, penetration depth, and the dislodgement rate of mucus by HBSS flow. In-silico : To conduct the numerical study, the viscosity and density of the mucus layer were initially measured. A power-law model was introduced to characterize the viscosity of the mucus. The fluid flow equations were solved and visualized using COMSOL software. The numerical approach was validated against experimental observations to ensure its accuracy

Article Snippet: The microfluidic device, visualized under a Zeiss Microscope (Axio Zoom V16) as shown in Fig. , was used for the observation of corresponding fluorescent particle movement within both the mucus and HBSS.

Techniques: In Vitro, Injection, Imaging, Microscopy, Software, In Silico, Viscosity

Journal: Drug Delivery and Translational Research

Article Title: Innovative microfluidic model for investigating the intestinal mucus barrier: numerical and experimental perspectives

doi: 10.1007/s13346-025-01818-8

Figure Lengend Snippet: Schematic representation of chip fabrication: ( A ) CAD drawing illustrating the microfluidic design with two parallel channels and interfacing pillars. ( B ) Photomask created from the CAD design for photolithography. ( C ) Fabricated microfluidic design on a silicon wafer by photolithography technique. ( D ) Digital microscope image (VHX-5000, Keyence Corp) used for quality assessment of the mold. White lines indicate the borders of regions with cured photoresist, verifying the quality of the photolithography step with well-created edges. ( E ) Final microfluidic chip made of PDMS, bonded to glass slides, and ready for in-vitro analysis

Article Snippet: The microfluidic device, visualized under a Zeiss Microscope (Axio Zoom V16) as shown in Fig. , was used for the observation of corresponding fluorescent particle movement within both the mucus and HBSS.

Techniques: Microscopy, In Vitro

Journal: Drug Delivery and Translational Research

Article Title: Innovative microfluidic model for investigating the intestinal mucus barrier: numerical and experimental perspectives

doi: 10.1007/s13346-025-01818-8

Figure Lengend Snippet: Experimental setup illustration: ( A ) Experimental setup comprising a Zeiss microscope for visualization of the fluorescent particles and an Elveflow pump for the injection of the HBSS into the microfluidic chip. ( B ) Time series of captured images aligned to the green channel (HBSS). ( C ) Time series of captured images aligned to the red channel (BSM). ( D ) Representation of the microfluidic chip filled with HPTS. ( E ) Thermal camera image showing the temperature of the mucus inside the chip just before the experiment began. ( F ) Illustration of the microfluidic chip area under microscopic observation using a 1 × lens at 45% zoom, focusing on the region of interest (ROI). This image was captured post-experiment with an HPTS solution injected (panel D ) to define the ROI. ( G ) Black and white mask used for image processing, derived from thresholding the mask shown in panel F

Article Snippet: The microfluidic device, visualized under a Zeiss Microscope (Axio Zoom V16) as shown in Fig. , was used for the observation of corresponding fluorescent particle movement within both the mucus and HBSS.

Techniques: Microscopy, Injection, Derivative Assay

Journal: PLoS ONE

Article Title: IFI16 Protein Mediates the Anti-inflammatory Actions of the Type-I Interferons through Suppression of Activation of Caspase-1 by Inflammasomes

doi: 10.1371/journal.pone.0027040

Figure Lengend Snippet: ( A ) Puromycin-resistant THP-1 cells either infected with control lentivirus (lanes 1 an2) or the virus expressing the shIFI16 RNA (lanes 3–8; cell populations from three different wells) were either left untreated (lanes 1, 3, 5, 7) or treated with IFN-α for 14 h. After treatment, total cell extracts containing increased amounts of proteins (∼100 µg/lane) were analyzed for the constitutive and induced levels of IFI16 and actin proteins. A long exposure was taken to detect the signal in all lanes. ( B ) Control THP-1 cells (lanes 1 and 2) or cell population from well # 9 (lanes 3 and 4) as shown in the panel (a) were either left untreated (lanes 1 and 3) or treated with IFN-α for 14 h (lanes 2 and 4). After the treatment, total RNA was analyzed for the steady-state levels of mRNA for the indicated genes. ( C ) Control THP-1 cells (lanes 1–4) or cells infected with virus expressing the shIFI16 mRNA (lanes 5–8; population # 9) were either left untreated (lanes 1 and 5) or treated with IFN-β (1,000 u/ml; lanes 2 and 6), LPS (100 ng/ml; lanes 3 and 7), or dsRNA (10 µg/ml; lanes 4 and 8) for 14 h. After the treatment, total cell lysates containing equal amounts of protein (∼100 µg/lane) were analyzed by immunoblotting using specific antibodies to the indicated proteins. FC, indicates the fold change in the levels of the activated caspase-1 (the p20 band) with respect to the control (lane 1). ( D ) Control THP-1 cells or cells infected with virus expressing the shIFI16 mRNA (population # 9) were either left untreated (white columns) or treated with IFN-β (1,000 u/ml; columns 2–5 and 7–10) for 14 h. After the treatment, total RNA levels were analyzed for the indicated genes by the quantitative TaqMan real-time PCR. The ratio of the test gene to actin mRNA was calculated in units (one unit being the ratio of the test gene to actin mRNA). Results are mean values of triplicate experiments and error bars represent standard deviation.

Article Snippet: THP-1 cells were either infected with control lentiviral particles (sc-108080; from Santa Cruz Biotech, Santa Cruz,, CA) or the lentiviral particles expressing shIFI16 RNA (sc-35633-V, Santa Cruz Biotech) in a six well plate as suggested by the supplier.

Techniques: Infection, Control, Virus, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Standard Deviation