Journal: Bioactive Materials

Article Title: Sustained release PLGA microspheres loaded with a bone-affinity Bmp2 enhance fracture healing and mitigate heterotopic ossification

doi: 10.1016/j.bioactmat.2026.02.050

Figure Lengend Snippet: Design and validation of bone-targeted Bmp2. a. Schematic representation of the Bmp2/D-Bmp2 plasmid constructs and the structure predicted by AlphaFold3. b. Immunofluorescence staining of HEK-293T cells transfected with the plasmids: phalloidin (green), DAPI (blue), and Alexa Fluor 647-conjugated anti-Flag antibodies (red). Scale bar: 10 μm. c. SDS-PAGE of purified proteins. d. Western blot validation of protein expression. e. Bmp2 activity reporter assay: schematic of the luciferase reporter system (left), representative fluorescence images, and statistical analysis of firefly luciferase activity by in vivo imaging system (IVIS) (a.u.: arbitrary units) (right, n = 3 per group). f. qPCR analysis of Bmp2 signaling pathway-related mRNA levels in MC3T3-E1 cells treated with the Bmp2 or D-Bmp2 protein (n = 3 per group). g, h. Western blot (g) and quantification of Bmp2 pathway-related protein expression in treated MC3T3-E1 cells (h) (n = 3 per group). i. Schematic of the HA-coated ELISA plate and HA-binding affinity assay (n = 3 per group). j. Tissue-specific binding assay of Bmp2 or D-Bmp2: Mouse muscle slices (left, scale bar: 100 μm) and undecalcified femur slices (right, scale bar: 200 μm) stained with AF647-conjugated anti-Flag antibodies. The data are presented as the means ± standard deviations (SDs). One-way ANOVA was used for multiple comparisons. Significance levels: ns (not significant), ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: After 24 h, the medium was changed to contain either Bmp2 or D-Bmp2 in DMEM, and the cells were further cultured for another 24 h. After the supernatant was removed, D-luciferin potassium salt (Beyotime, China) was added, and the firefly fluorescence signal was recorded using an in vivo imaging system (IVIS) (PerkinElmer, USA).

Techniques: Biomarker Discovery, Plasmid Preparation, Construct, Immunofluorescence, Staining, Transfection, SDS Page, Purification, Western Blot, Expressing, Activity Assay, Reporter Assay, Luciferase, Fluorescence, In Vivo Imaging, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Materials Today Bio

Article Title: Cholesterol analogs modulate lipid nanoparticle performance for mRNA delivery after lyophilization and enable ocular disease therapy

doi: 10.1016/j.mtbio.2026.103044

Figure Lengend Snippet: In vivo biodistribution and bioluminescence quantification of mRNA( FLuc )-LNPs containing various cholesterol analogs and ionizable lipids. (A) Schematic overview. (B–C) Representative IVIS images of C57BL/6J mice 6 h after intravenous (i.v.) injection of freshly prepared (B) or lyophilized and reconstituted (C) mRNA-LNPs formulated with either MC3 or ALC-0315 and different cholesterol analogs. (D–G) Quantification of average radiance (photons/s/cm 2 /sr) in the spleen, liver, and lungs of mice treated with freshly prepared (D and F) or lyophilized and reconstituted (E and G) MC3-based mRNA-LNPs (D and E) and ALC-0315-based mRNA-LNPs (F and G). (H–K) Normalized average radiance values in each organ from mice receiving C24-alkyl-substituted cholesterol analog-containing MC3-based mRNA-LNPs (H and I) and ALC-0315-based mRNA-LNPs (J and K), which were freshly prepared (H and J) or lyophilized and reconstituted (I and K), expressed relative to cholesterol-based LNPs. The data are shown as the means ± s.d.s ( n = 3 biologically independent mice per group). Statistical significance was evaluated via two-way ANOVA followed by Tukey's post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Bioluminescence signals from these tissues were immediately captured via the IVIS® in vivo imaging system (PerkinElmer, Waltham, MA, USA) and quantified with Living Image Software v4.7.4.

Techniques: In Vivo, Injection