Journal: Journal of Nanobiotechnology

Article Title: Neutrophil membrane-coated and MMP2-responsive nanoparticles deliver PIM1 inhibitor to alleviate inflammatory arthritis through inhibiting Th17 cell differentiation

doi: 10.1186/s12951-026-04280-x

Figure Lengend Snippet: Preparation and characterization of NM@MRP-NP ( A ) Morphology of MRP-NP, NM and NM@MRP-NP observed by SEM. Scale bar = 100 nm. ( B - D ) Size distributions ( B ), Z-average diameter ( C ) and zeta potential ( D ) of MRP-NP, NM and NM@MRP-NP measured by DLS ( n = 5). ( E ) Z-average diameter of MRP-NP, NM and NM@MRP-NP after incubation with mouse serum or FBS for different durations ( n = 5). ( F ) Fluorescence image of NM@MRP-NP loaded with different concentrations of Cy5.5. ( G ) Drug release rates of MRP-NP and NM@MRP-NP with or without MMP2 ( n = 5)

Article Snippet: The resulting NM@MRP-NP was collected by centrifugation at 20,000×g at 4 °C for 20 min. For the fluorescence tracing experiment, Cy5.5 (MCE, #HY-D0924) was substituted for AZD1208 and encapsulated into the nanoparticles.

Techniques: Zeta Potential Analyzer, Incubation, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Neutrophil membrane-coated and MMP2-responsive nanoparticles deliver PIM1 inhibitor to alleviate inflammatory arthritis through inhibiting Th17 cell differentiation

doi: 10.1186/s12951-026-04280-x

Figure Lengend Snippet: Biological characteristics of NM@MRP-NP. ( A ) Western blot analysis of the expression of CD11a, CD11b and L-selectin on the neutrophil membrane before and after LPS treatment ( n = 9). ( B ) Representative immunofluorescence images of CD11a, CD11b and L-selectin on the neutrophil membrane before and after LPS treatment. Scale bar = 20 μm. ( C - D ) Western blot analysis ( C ) and immunofluorescence staining ( D ) confirmed the transfer of CD11a, CD11b and L-selectin to the NM@MRP-NP. Scale bar = 100 μm. ( E - F ) Uptake of Cy5.5 by activated or unactivated CD4 + T cells treated with vehicle, MRP-NP, NM@MRP-NP or NM@MRP-NP+MMP2 ( n = 9). The statistical analyses were performed as follows: paired t test (A, F) and one-way ANOVA followed by Bonferroni’s post hoc comparisons test ( F ). ** indicates p < 0.01, *** indicates p < 0.001, **** indicates p < 0.0001, and n.s. indicates no statistical significance

Article Snippet: The resulting NM@MRP-NP was collected by centrifugation at 20,000×g at 4 °C for 20 min. For the fluorescence tracing experiment, Cy5.5 (MCE, #HY-D0924) was substituted for AZD1208 and encapsulated into the nanoparticles.

Techniques: Western Blot, Expressing, Membrane, Immunofluorescence, Staining

Journal: Journal of Nanobiotechnology

Article Title: Neutrophil membrane-coated and MMP2-responsive nanoparticles deliver PIM1 inhibitor to alleviate inflammatory arthritis through inhibiting Th17 cell differentiation

doi: 10.1186/s12951-026-04280-x

Figure Lengend Snippet: Targeting effect of NM@MRP-NP in vivo. ( A ) Cy5.5 fluorescence images of the inflamed ankles of SKG mice at different time points after treatment with vehicle, free Cy5.5, Cy5.5-labeled MRP-NP or NM@MRP-NP. ( B ) Cy5.5 fluorescence images of synovial fluid collected from SKG mice 8 h after treatment with vehicle, free Cy5.5, Cy5.5-labeled MRP-NP or NM@MRP-NP. ( C ) Cy5.5 fluorescence images of the ankles and the major organs dissected from SKG mice 8 h after treatment with vehicle, free Cy5.5, Cy5.5-labeled MRP-NP or NM@MRP-NP. ( D ) Immunofluorescence images of Cy5.5 and CD4 + T cells in the inflamed joints of SKG mice treated with vehicle, free Cy5.5, Cy5.5-labeled MRP-NP or NM@MRP-NP. Scale bar = 50 μm. ( E ) Quantitative analysis of the Cy5.5 fluorescent signal in the inflamed ankles of SKG mice at different time points after receiving Cy5.5-labeled NM@MRP-NP ( n = 6). ( F ) Quantitative analysis of Cy5.5 fluorescent signal in synovial fluid collected from SKG mice 8 h after treatment with vehicle, free Cy5.5, Cy5.5-labeled MRP-NP or NM@MRP-NP ( n = 5). ( G ) Quantitative analysis of Cy5.5 fluorescence signal in the ankles and the major organs dissected from SKG mice 8 h after treatment with vehicle, free Cy5.5, Cy5.5-labeled MRP-NP or NM@MRP-NP ( n = 5). ( H ) Line-scan analysis of the colocalization of Cy5.5 and CD4 + T cells in the immunofluorescence images in Fig. 7D. The statistical analyses were performed as follows: one-way ANOVA followed by Bonferroni’s post hoc comparisons test (F, G). **** indicates p < 0.0001

Article Snippet: The resulting NM@MRP-NP was collected by centrifugation at 20,000×g at 4 °C for 20 min. For the fluorescence tracing experiment, Cy5.5 (MCE, #HY-D0924) was substituted for AZD1208 and encapsulated into the nanoparticles.

Techniques: In Vivo, Fluorescence, Labeling, Immunofluorescence

Journal: Theranostics

Article Title: Engineering blood exosomes for tumor-targeting efficient gene/chemo combination therapy

doi: 10.7150/thno.45028

Figure Lengend Snippet: Characterization of D-Exos/miR21i-L17E. (A, B) The particle size distribution and representative TEM images of SMNC-Exos and D-Exos/miR21i-L17E, respectively. (scale bar: 20 nm) (C) Western blots analysis of specific Tf receptors and exosome marker proteins (CD63, CD81 and CD9) of SMNC-Exos, D-Exos/miR21i and D-Exos/miR21i-L17E. (D) Detection of fluorescent signals in purified SMNC-Exos, D-Exos, D-Exos/miR21i and D-Exos/miR21i-L17E at different excitation/emission (Ex/Em) wavelength. The SMNC-Exos and chol-miR21i contained in each sample were labeled by FITC and Cy5, respectively. (E) UV-vis spectra of the SMNC-Exos, Dox, chol-miR21i-Cy5, L17E and D-Exos/miR21i-L17E. The inset shows the magnified UV-vis spectra from 600 to 700 nm. (F) Gel retention assay of D-Exos/miR21i prepared with D-Exos and varied concentrations of cholesterol-conjugated miR-21i or non-cholesterol-conjugated miR-21i. The Cy5-labeled miR-21i bands are displayed. (G) Schematic of the methods for confirming the localization of chol-miR21i on D-Exos/miR21i. Cy5-labeled miR-21 is complementary to chol-miR21i on D-Exos/miR21i. If miR-21i is completely display on the surface of exosomes, Cy5-miR21 will bind to D-Exos/miR21i, otherwise it will not. (H) Analysis for the binding of Cy5-miR21 to D-Exos/miR21i by gel electrophoresis. The Cy5-labeled miR-21 bands are displayed here. (I) Release profiles of Dox from D-Exos/miR21i-L17E in PBS (pH=7.4) and acetate buffer (pH=5.0) for 24 h. Data in (I) are presented as the mean ± standard deviation (SD) from three independent experiments (n=3).

Article Snippet: Cy5 and Cy5.5 fluorescence dyes were purchased from US Everbright ® Inc (Suzhou, China).

Techniques: Western Blot, Marker, Purification, Labeling, Binding Assay, Nucleic Acid Electrophoresis, Standard Deviation

Journal: Theranostics

Article Title: Engineering blood exosomes for tumor-targeting efficient gene/chemo combination therapy

doi: 10.7150/thno.45028

Figure Lengend Snippet: Cellular uptake and intracellular distribution of D-Exos/miR21i-L17E. (A) Confocal images of U87 cells incubated with D-Exos/miR21i and D-Exos/miR21i-L17E for different times (1 h, 2 h, 4 h). The fluorescence spots of Dox (red) and chol-miR21i-Cy5 (purple) were observed. The nucleus and endo/lysosome were stained with DAPI (blue) and Lysotracker Green (green), respectively. (B) Co-localization ratio of the miR-21i signals (purple) in D-Exos/miR21i and D-Exos/miR21i-L17E to endosome signals (green) at different time points. (C) Flow cytometry profile of U87 cells incubated with free Dox and chol-miR21i, D-Exos, Exos/miR21i, D-Exos/miR21i and D-Exos/miR21i-L17E for 4 h. The y-axis represents cell counts and the x-axis represents the Dox and chol-miR21i-Cy5 fluorescence intensity, respectively. Data in (B) are presented as the mean ± SD (n>50) from three independent experiments (n=3). The significant levels are shown as ** p < 0.01, *** p < 0.001.

Article Snippet: Cy5 and Cy5.5 fluorescence dyes were purchased from US Everbright ® Inc (Suzhou, China).

Techniques: Incubation, Fluorescence, Staining, Flow Cytometry

Journal: Theranostics

Article Title: Engineering blood exosomes for tumor-targeting efficient gene/chemo combination therapy

doi: 10.7150/thno.45028

Figure Lengend Snippet: Tumor targeting efficiency of engineered exosomes in tumor-bearing mice in vivo . (A) In vivo NIRF imaging of U87 tumor-bearing nude mice after intravenous injection of Cy5.5-labeled D-Exos/miR21i-L17E with/without an external MF at 1 h, 4 h and 24 h post-injection. (B) Quantitative analysis of the tumor accumulation of the D-Exos/miR21i-L17E based on the fluorescence intensity from the in vivo images. (C) The accumulation of Dox and Cy5-miR21i in the tumor section, where red represents Dox, green represents miR21i and cell nucleus was stained by DAPI. The scale bar is 100 μm. Data in (B) are presented as the mean ± SD (n=5).

Article Snippet: Cy5 and Cy5.5 fluorescence dyes were purchased from US Everbright ® Inc (Suzhou, China).

Techniques: In Vivo, Imaging, Injection, Labeling, Fluorescence, Staining