Journal: Biomaterials Research

Article Title: Autotaxin-Scavenging Nanoliposomes for Prolonged Colon Retention and Autophagy-Mediated Mucosal Immune Restoration in Colitis

doi: 10.34133/bmr.0345

Figure Lengend Snippet: Characterization and ATX-scavenging activity of AS-Lipo@R. (A) Cryo-TEM images of Lipo and AS-Lipo (scale bar, 100 nm). (B) Particle size and zeta potential of Lipo and AS-Lipo measured by DLS. (C) Colloidal stability of Lipo and AS-Lipo was evaluated by monitoring changes in particle size and zeta potential over 72 h in PBS using DLS. (D) Concentration-dependent inhibition of recombinant mouse ATX activity as a function of BMP-22 concentration in free BMP-22, Lipo, and AS-Lipo, evaluated using a choline release assay. (E) ATX binding to the surface of Lipo and AS-Lipo as a function of ATX concentration, quantified by ELISA. (F) HPLC chromatograms showing rapamycin (R) encapsulation in AS-Lipo@R, as indicated by the disappearance of the free R peak. (G) Cumulative release profiles of rapamycin from Lipo@R and AS-Lipo@R in PBS over 72 h measured by HPLC. (H) Fluorescence imaging showing colocalization of DiO-labeled Lipo or AS-Lipo (green), Alexa Fluor 647 (AF647)-labeled ATX (red), and LysoTracker (purple) in RAW 264.7 macrophages. AS-Lipo-bound ATX is internalized and colocalizes with lysosomes, indicating lysosomal degradation. Right panels show fluorescence intensity profiles along the indicated lines, quantifying the colocalization (scale bar, 100 and 50 μm). Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Cryo-TEM micrographs were acquired using a JEM-2100F electron microscope (JEOL, Japan).

Techniques: Activity Assay, Zeta Potential Analyzer, Concentration Assay, Inhibition, Recombinant, Release Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Encapsulation, Fluorescence, Imaging, Labeling