nanofibril exposure  (MedChemExpress)

Journal: Bioactive Materials

Article Title: Food-derived β-lactoglobulin nanofibrils: An efficacy, safe, and scalable solution to overcome oral insulin delivery challenges

doi: 10.1016/j.bioactmat.2025.11.020

Figure Lengend Snippet: Schematic illustration of BLG nanofibril-mediated oral peptide delivery. (a) Preparation of nanofibrils. BLG, a whey protein byproduct of cheese production, is converted into nanofibrils through thermal treatment. (b – c) Benefits and mechanism of nanofibril-mediated oral peptide delivery. Nanofibrils induce transient intestinal permeability enhancement through calcium influx-mediated calpain activation. The gastrointestinal degradability of nanofibrils ensures complete restoration of intestinal barrier function post-delivery.

Article Snippet: To investigate molecular mechanisms, monolayers were pre-treated for 2 h with specific inhibitors prior to nanofibril exposure: 500 nM MLCK inhibitor peptide 18 (MLCK inhibitor), 50 μM MDL-28170 (Calpain inhibitor), 200 nM Bisindolymaleimide I (PKC inhibitor), 10 μM Y-27632 (ROCK inhibitor), 5 μM Adezmapimod (MAPK inhibitor) (all inhibitors were purchased from MCE).

Techniques: Permeability, Activation Assay

Journal: Bioactive Materials

Article Title: Food-derived β-lactoglobulin nanofibrils: An efficacy, safe, and scalable solution to overcome oral insulin delivery challenges

doi: 10.1016/j.bioactmat.2025.11.020

Figure Lengend Snippet: Comparative evaluation of mucosal penetration and intestinal permeability modulation by BLG monomers versus BLG nanofibrils . (a – c) Mucus penetration characteristics: (a) 3D confocal microscopy images showing distribution of FITC-labeled monomers (green) vs nanofibrils (green) in jejunal mucus (red, stained with rhodamine-WGA) 30 min post-administration; (b) Representative particle trajectories and (c) mean squared displacement (MSD) analysis in stimulated intestinal mucus. (d – f) In vivo permeability enhancement: (d) Experimental workflow; (e) Time-dependent enhancement of 4 kDa FITC-dextran absorption following nanofibril pretreatment, n = 3; (f) Molecular weight dependence of permeability enhancement, n = 3. (g – i) In vitro reversible permeability modulation: (g) Experimental setup of Caco-2 monolayer; (h) Enhanced transport of 4 kDa FITC-dextran for nanofibril-treated monolayers, n = 3; (i) Real-time TEER monitoring demonstrating transient barrier disruption and recovery for nanofibril treatment, n = 3. Significance levels: ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: To investigate molecular mechanisms, monolayers were pre-treated for 2 h with specific inhibitors prior to nanofibril exposure: 500 nM MLCK inhibitor peptide 18 (MLCK inhibitor), 50 μM MDL-28170 (Calpain inhibitor), 200 nM Bisindolymaleimide I (PKC inhibitor), 10 μM Y-27632 (ROCK inhibitor), 5 μM Adezmapimod (MAPK inhibitor) (all inhibitors were purchased from MCE).

Techniques: Permeability, Confocal Microscopy, Labeling, Staining, In Vivo, Molecular Weight, In Vitro, Disruption

Journal: Bioactive Materials

Article Title: Food-derived β-lactoglobulin nanofibrils: An efficacy, safe, and scalable solution to overcome oral insulin delivery challenges

doi: 10.1016/j.bioactmat.2025.11.020

Figure Lengend Snippet: Oral insulin delivery efficacy in mice. (a) Experimental scheme: Fasting mice received oral gavage of 200 μL either PBS, 20 mg/mL monomer solution, or 20 mg/mL nanofibril solution, followed by oral administration of insulin capsules (20 IU/kg) after 2 h. A pharmacological control group received subcutaneous insulin injection (2 IU/kg). (b – d) Healthy mice: (b) Blood glucose levels, (c) area under the curve (AUC) of blood glucose, and (d) serum insulin levels. (e – g) Severe diabetes model: (e) Blood glucose levels, (f) AUC of blood glucose, and (g) serum insulin levels. (h – i) Mild diabetes model: (h) Blood glucose levels and (i) AUC of blood glucose. Data present mean ± SEM (n = 6 for glucose analysis; n = 3 for serum insulin). Significance levels: ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: To investigate molecular mechanisms, monolayers were pre-treated for 2 h with specific inhibitors prior to nanofibril exposure: 500 nM MLCK inhibitor peptide 18 (MLCK inhibitor), 50 μM MDL-28170 (Calpain inhibitor), 200 nM Bisindolymaleimide I (PKC inhibitor), 10 μM Y-27632 (ROCK inhibitor), 5 μM Adezmapimod (MAPK inhibitor) (all inhibitors were purchased from MCE).

Techniques: Capsules, Control, Injection

Journal: Bioactive Materials

Article Title: Food-derived β-lactoglobulin nanofibrils: An efficacy, safe, and scalable solution to overcome oral insulin delivery challenges

doi: 10.1016/j.bioactmat.2025.11.020

Figure Lengend Snippet: Reversible modulation of tight junctions by BLG nanofibrils in vivo. (a) Experimental timeline showing nanofibril administration and jejunal tissue collection for tight junction analysis. (b – d) Dynamics of tight junctions in the jejunum following nanofibril gavage: (b) TEM images, TJ (tight junction), AJ (adherens junction); (c) Western blot analysis of ZO-1, occludin, and claudin-1 levels; (d) Densitometric analysis. (e – g) Comparison of tight junctions after treatment with PBS, monomers, or nanofibrils: (e) TEM images; (f) Western blot analysis; (g) Densitometric analysis. (h – k) Immunofluorescence assessment of tight junction proteins following nanofibril gavage: (h) Representative fluorescence images (tight junction proteins: red; nuclei: blue); (i–k) Fluorescence intensity analysis. Data present mean ± SEM (n = 3). Significance levels: ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001.

Article Snippet: To investigate molecular mechanisms, monolayers were pre-treated for 2 h with specific inhibitors prior to nanofibril exposure: 500 nM MLCK inhibitor peptide 18 (MLCK inhibitor), 50 μM MDL-28170 (Calpain inhibitor), 200 nM Bisindolymaleimide I (PKC inhibitor), 10 μM Y-27632 (ROCK inhibitor), 5 μM Adezmapimod (MAPK inhibitor) (all inhibitors were purchased from MCE).

Techniques: In Vivo, Western Blot, Comparison, Immunofluorescence, Fluorescence

Journal: Bioactive Materials

Article Title: Food-derived β-lactoglobulin nanofibrils: An efficacy, safe, and scalable solution to overcome oral insulin delivery challenges

doi: 10.1016/j.bioactmat.2025.11.020

Figure Lengend Snippet: Nanofibrils modulate tight junctions through Ca 2+ influx-mediated calpain activation. ( a) Immunofluorescence analysis of cytoskeletal integrity following 2-h treatment with PBS, monomers, or nanofibrils. F-actin filaments (red, phalloidin) and nuclei (blue, DAPI). (b – d) Intracellular Ca 2+ levels: (b) Intracellular Ca 2+ levels following 2-h treatment with PBS, monomers, or nanofibrils; (c) Quantification of relative Fluo-4 fluorescence intensity; (d) Time-lapse tracking of intracellular Ca 2+ levels with the stimulation of 2 mM extracellular Ca 2+ . (e – f) FRAP analysis of cell membrane fluidity: (e) Representative fluorescence images of pre-bleaching and recovery; (f) Normalized recovery kinetics. (g) Fluorescence imaging and (h) quantitative analysis of cells stained with DiBAC 4 (3) under nanofibril treatment. (i – j) Pharmacological inhibitors: (i) Experimental workflow; (j) Attenuation of nanofibril-induced permeability by calpain inhibitor. Data present mean ± SEM (n = 3). Significance levels: ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: To investigate molecular mechanisms, monolayers were pre-treated for 2 h with specific inhibitors prior to nanofibril exposure: 500 nM MLCK inhibitor peptide 18 (MLCK inhibitor), 50 μM MDL-28170 (Calpain inhibitor), 200 nM Bisindolymaleimide I (PKC inhibitor), 10 μM Y-27632 (ROCK inhibitor), 5 μM Adezmapimod (MAPK inhibitor) (all inhibitors were purchased from MCE).

Techniques: Activation Assay, Immunofluorescence, Fluorescence, Membrane, Imaging, Staining, Permeability