Journal: Biomedicines

Article Title: Therapeutic Exosomes Carrying VEGFA siRNA Inhibit Pathological Corneal Angiogenesis via PI3K–Akt–Caspase-3 Signaling

doi: 10.3390/biomedicines14010246

Figure Lengend Snippet: VEGFA siRNA-FAM-loaded exosome-mediated introduction into MSCs. ( A ) VEGFA siRNA-FAM (green fluorescence) was successfully injected into exosomes and delivered to target cells. White circles represent exosomes fluorescently labeled with VEGFA siRNA-FAM. ( B ) RT-PCR and Western Blot analysis of MSCs over time after treatment. A decrease in VEGFA mRNA and protein expression was observed under the same conditions. ( C ) Intracellular and extracellular VEGFA levels in MSCs were quantified over time using a multiplex assay. Bars represent 200 μm. * p < 0.05 and ** p < 0.01 were considered significant differences.

Article Snippet: After establishing the model, mice ( n = 6 per group) were randomly assigned to five groups: Normal control group (no suturing and injection; Normal), a Bevacizumab-treated group (Bevacizumab), a VEGFA siRNA-FAM-loaded exosome-treated group ( VEGFA siRNA-FAM-EXO), a non-loaded Exosome-treated group (Exosome), and a PBS-treated control group (PBS).

Techniques: Fluorescence, Injection, Labeling, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Multiplex Assay

Journal: Biomedicines

Article Title: Therapeutic Exosomes Carrying VEGFA siRNA Inhibit Pathological Corneal Angiogenesis via PI3K–Akt–Caspase-3 Signaling

doi: 10.3390/biomedicines14010246

Figure Lengend Snippet: Decreased cell viability, increased apoptosis, and inhibited HUVEC-derived tube formation in MSCs induced by VEGFA siRNA-FAM-loaded exosomes in vitro. ( A ) Cell viability (CCK 8) as a function of VEGFA siRNA-FAM-loaded exosomes into the target cells according to exposure time. ( B ) Time-dependent effects of VEGFA siRNA treatment as determined by FACS analysis. Terms: Q1, necrosis; Q2, late apoptosis; Q3, survival; Q4, early apoptosis. ( C ) Morphological changes in tube formation in HUVECs over time after VEGFA siRNA-FAM-loaded exosome treatment. ( D ) Quantitative graph of the number of master segments involved in tube formation and total master segment length. Statistics: * p < 0.05 and *** p < 0.001 were considered significant differences, and were evaluated by one-way ANOVA followed by Tukey’s multiple comparisons. Scale bar = 200 μm.

Article Snippet: After establishing the model, mice ( n = 6 per group) were randomly assigned to five groups: Normal control group (no suturing and injection; Normal), a Bevacizumab-treated group (Bevacizumab), a VEGFA siRNA-FAM-loaded exosome-treated group ( VEGFA siRNA-FAM-EXO), a non-loaded Exosome-treated group (Exosome), and a PBS-treated control group (PBS).

Techniques: Derivative Assay, In Vitro, CCK-8 Assay

Journal: Biomedicines

Article Title: Therapeutic Exosomes Carrying VEGFA siRNA Inhibit Pathological Corneal Angiogenesis via PI3K–Akt–Caspase-3 Signaling

doi: 10.3390/biomedicines14010246

Figure Lengend Snippet: Establishment of conditions for the treatment of VEGFA siRNA-FAM-loaded exosomes in the right eye of each mouse with the CNV model in vivo. ( A ) Experimental timeline. After successful establishment of the CNV model, intrastromal injections were performed into the corneal stroma on days 7, 9, 12, 14, and 17, and sacrifice and biopsy were performed on day 21. ( B ) Fluorescence imaging after treatment with VEGFA siRNA-FAM-loaded exosomes. Fluorescence images of the VEGFA siRNA-FAM-EXO-treated group are shown. Green signal (FAM) indicates that VEGFA siRNA-loaded exosomes have successfully entered the corneal stroma. ( C ) Quantification of total radiative efficiency (TRE) and average radiative efficiency (AER) at 0, 12, 24, 48, and 72 h after VEGFA siRNA-FAM-EXO treatment. Data are presented as mean ± standard error. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test. Standard criteria: * p < 0.05, ** p < 0.01.

Article Snippet: After establishing the model, mice ( n = 6 per group) were randomly assigned to five groups: Normal control group (no suturing and injection; Normal), a Bevacizumab-treated group (Bevacizumab), a VEGFA siRNA-FAM-loaded exosome-treated group ( VEGFA siRNA-FAM-EXO), a non-loaded Exosome-treated group (Exosome), and a PBS-treated control group (PBS).

Techniques: In Vivo, Fluorescence, Imaging, Comparison

Journal: Biomedicines

Article Title: Therapeutic Exosomes Carrying VEGFA siRNA Inhibit Pathological Corneal Angiogenesis via PI3K–Akt–Caspase-3 Signaling

doi: 10.3390/biomedicines14010246

Figure Lengend Snippet: VEGFA siRNA-FAM-loaded exosomes inhibit corneal neovascularization in vivo. ( A ) Representative anterior segment images from each group (Normal, Bevacizumab, VEGFA siRNA-FAM-EXO, Exosomes, PBS). Images were taken on days 7, 9, 12, 14, 17, and 19 after model induction to compare changes in neovascularization over time. ( B ) Quantitative results of the angiogenic index (AI) (mean ± standard error, n = 6). A significant decrease was observed in the VEGFA siRNA-FAM-EXO group from days 14 to 19. ( C ) Optical photograph (top) and immunohistochemical staining (bottom) of mouse corneas. The expression levels of CD31 and VEGFA were indicated by their respective fluorescence (green and red) within circles (corneas), and vascular signals were confirmed in the expanded image. ( D ) Western Blot analysis. The expression of CD31 (a marker of vascular endothelial cells) and VEGFA was compared in lysates collected from corneal tissues at day 21 (the time of sacrifice) in five groups. * p < 0.05 and ** p < 0.01 were considered significant differences. Scale bar = 200 μm.

Article Snippet: After establishing the model, mice ( n = 6 per group) were randomly assigned to five groups: Normal control group (no suturing and injection; Normal), a Bevacizumab-treated group (Bevacizumab), a VEGFA siRNA-FAM-loaded exosome-treated group ( VEGFA siRNA-FAM-EXO), a non-loaded Exosome-treated group (Exosome), and a PBS-treated control group (PBS).

Techniques: In Vivo, Immunohistochemical staining, Staining, Expressing, Fluorescence, Western Blot, Marker

Journal: Biomedicines

Article Title: Therapeutic Exosomes Carrying VEGFA siRNA Inhibit Pathological Corneal Angiogenesis via PI3K–Akt–Caspase-3 Signaling

doi: 10.3390/biomedicines14010246

Figure Lengend Snippet: VEGFA siRNA-FAM-loaded exosomes reduce angiogenesis and attenuate inflammatory changes. ( A ) H&E-stained mouse corneal tissue images. Anterior segment sections (top) and enlarged corneal sections (bottom) are shown for each group: Normal, Bevacizumab, VEGFA siRNA-FAM-EXO, Exosome, and PBS. Neovascular invasion and inflammatory hypertrophy were attenuated in the VEGFA siRNA-FAM-EXO group. Scale bar = 200 μm. ( B ) Quantification of the number of corneal neovascularization (mean ± standard error, n = 4). A significant reduction in blood vessels was observed in the VEGFA siRNA-FAM-EXO group compared to PBS. ( C ) Quantification of corneal epithelial thickness (mean ± standard error, n = 6). Inflammatory epithelial hyperplasia was suppressed in the VEGFA siRNA-FAM-EXO group. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons, and the significance levels were * p < 0.05 and ** p < 0.01.

Article Snippet: After establishing the model, mice ( n = 6 per group) were randomly assigned to five groups: Normal control group (no suturing and injection; Normal), a Bevacizumab-treated group (Bevacizumab), a VEGFA siRNA-FAM-loaded exosome-treated group ( VEGFA siRNA-FAM-EXO), a non-loaded Exosome-treated group (Exosome), and a PBS-treated control group (PBS).

Techniques: Staining

Journal: Biomedicines

Article Title: Therapeutic Exosomes Carrying VEGFA siRNA Inhibit Pathological Corneal Angiogenesis via PI3K–Akt–Caspase-3 Signaling

doi: 10.3390/biomedicines14010246

Figure Lengend Snippet: VEGFA siRNA-FAM-loaded exosomes inhibit PI3K/Akt signaling and induce apoptosis. ( A ) Western Blot analysis showed that the VEGFA siRNA-FAM-EXO group showed decreased levels of phosphorylated PI3K (p-PI3K, 85 kDa) and phosphorylated Akt (p-Akt, 60 kDa), while cleaved caspase 3 (15–17 kDa) increased. Total PI3K (85 kDa) and total Akt (60 kDa) were presented as comparative indices, and β-actin (45 kDa) was used as a loading control. Band intensities were quantified by normalization to β-actin. ( B ) Immunofluorescence staining analysis. The expression of p-PI3K (green), p-Akt (red), and cleaved caspase-3 (green) in each treatment group (Normal, Bevacizumab, VEGFA siRNA-FAM-EXO, Exosome, and PBS) was presented as a merged image with DAPI (blue) nuclear staining. The VEGFA siRNA-FAM-EXO group showed a decrease in p-PI3K and p-Akt signals and an increase in cleaved caspase-3 signals, consistent with the Western Blot results. The white scale bar represents 200 μm.

Article Snippet: After establishing the model, mice ( n = 6 per group) were randomly assigned to five groups: Normal control group (no suturing and injection; Normal), a Bevacizumab-treated group (Bevacizumab), a VEGFA siRNA-FAM-loaded exosome-treated group ( VEGFA siRNA-FAM-EXO), a non-loaded Exosome-treated group (Exosome), and a PBS-treated control group (PBS).

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

Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: Validation of M2-Exo stimulated FAM83A-mediated EMT activation through FAM83A silencing and examining by RT-qPCR analysis. ( A ) Silencing of FAM83A in M2-Exo–induced HT-29 cells using varying concentrations of FAM83A-siRNA (sir10, 20, 50 nM) and corresponding scramble-siRNA controls (scr10, 20, 50 nM). HT-29 cells alone and HT-29 cells stimulated with M2-Exo served as negative and positive controls, respectively. Silencing FAM83A attenuates M2-Exo–mediated EMT, as demonstrated by reduced expression of the ( B ) mesenchymal marker N-Cadherin and ( C ) the metastasis marker MMP-2, in M2-Exo stimulated HT-29 cells, respectively. ( D ) FAM83A expression was assessed in breast cancer (MCF-7), melanoma (451LU), and glioblastoma (T98G) cells without M2-Exosome stimulation (NS = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001). Western Blot images obtained with N-cadherin and E-cadherin to assess the response of HT-29 cells against M2-Exo, M2-Exo–Akt inhibitor co-load, M2-Exo–siRNA co-load, and M2-Exo–Akt inhibitor-siRNA co-load. RT-qPCR experiments were conducted in triplicate across three biological replicates. Western blot analysis was performed in duplicate using protein samples pooled from three independent biological replicates.

Article Snippet: Silencing FAM83A attenuates M2-Exo–mediated EMT, as demonstrated by reduced expression of the ( B ) mesenchymal marker N-Cadherin and ( C ) the metastasis marker MMP-2, in M2-Exo stimulated HT-29 cells, respectively. ( D ) FAM83A expression was assessed in breast cancer (MCF-7), melanoma (451LU), and glioblastoma (T98G) cells without M2-Exosome stimulation (NS = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001).

Techniques: Biomarker Discovery, Activation Assay, Quantitative RT-PCR, Expressing, Marker, Western Blot