Journal: Materials Today Bio

Article Title: Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma

doi: 10.1016/j.mtbio.2024.101244

Figure Lengend Snippet: Barrier permeation enhancement effect of EVs and efficient delivery of aPD-L1/EV into brain. ( A ) Scheme of the in vitro barrier permeation test with C2BBe1 trans-well system (created from Biorender.com ). ( B ) Amounts of permeated aPD-L1 with free aPD-L1 or aPD-L1/EV administration through the C2BBe1 barrier. ( C ) Comparison of drug delivery efficiency with RO or IN administration of FITC-conjugated IgG or IgG/EV. The upper column displays the fluorescence of IgG within brain sections, and the lower column shows the cumulative fluorescence intensity of brain sections. ( D ) Distribution of aPD-L1 in GL261 tumor-bearing mice. Green fluorescence indicates aPD-L1, and nuclei were stained with DAPI. Statistics in ( B, C ) were conducted by one-way ANOVA with Bonferroni post hoc test. Asterisks represent a significant difference between indicated groups, with significance defined as p < 0.05. Data were presented as mean ± SD with n = 3.

Article Snippet: The GL261-luc murine glioblastoma cell line was purchased from Creative Biolabs (Shirley, NY, USA).

Techniques: In Vitro, Comparison, Fluorescence, Staining

Journal: Materials Today Bio

Article Title: Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma

doi: 10.1016/j.mtbio.2024.101244

Figure Lengend Snippet: Therapeutic effect of ICI/EVs in managing orthotopic GL261 tumors. ( A ) Schematic of experimental procedures for GL261 tumor management (created from Biorender.com ). ( B, C ) Tumor volumes were evaluated with IVIS, and representative images are shown in ( B ). Changes in tumor volume were calculated based on the total flux observed on day 0 ( C ). ( D ) The survival of orthotopic GL261 tumor-bearing mice post-treatment was analyzed. The aPD-L1/EV + aCTLA-4/EV group demonstrated an extended survival time compared to other treatment groups. ( E ) Ki67 staining of GL261 tumors. Scale bar: 50 μm. Statistics for ( C ) were obtained from GLM with Bonferroni post hoc test, and for ( D ) from the Kaplan-Meier survival analysis with log rank test. The asterisk in ( C ) indicates a significant difference from the PBS and aPD-L1 treatments. In ( D ), the asterisk and hash signs represent significant difference from all treatment groups and from the PBS group, respectively. Significance was defined as p < 0.05. Data are presented as mean ± SEM with n = 4 in ( C ) and in ( D ) with n = 8∼18.

Article Snippet: The GL261-luc murine glioblastoma cell line was purchased from Creative Biolabs (Shirley, NY, USA).

Techniques: Staining

Journal: Materials Today Bio

Article Title: Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma

doi: 10.1016/j.mtbio.2024.101244

Figure Lengend Snippet: Immune cells in GL261 tumors post treatment. ( A ) IHC images of cytotoxic T cells within the tumor tissue. Green fluorescence indicates CD8 expression on T cells, while nuclei are counterstained with DAPI. Scale bar: 50 μm. ( B ) IHC images of M1 macrophages within tumor tissue. Green fluorescence indicates F4/80-stained macrophages, and red fluorescence indicates iNOS expression. Nuclei are counterstained with DAPI. Scale bar: 20 μm. ( C-E ) Flow cytometry analysis results for total T cells ( C ), cytotoxic T cells ( D ), and M1 macrophages ( E ). Statistics for ( C, D, E ) were obtained from one-way ANOVA with Bonferroni post hoc test, with significance defined as p < 0.05. Data are expressed as mean ± SEM with n = 5.

Article Snippet: The GL261-luc murine glioblastoma cell line was purchased from Creative Biolabs (Shirley, NY, USA).

Techniques: Fluorescence, Expressing, Staining, Flow Cytometry