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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a <t>fluorescein</t> <t>isothiocyanate</t> <t>(FITC)</t> dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.
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CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a fluorescein isothiocyanate (FITC) dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.

Journal: Journal of Pharmaceutical Analysis

Article Title: Hyaluronic acid-modified polymeric nanoplatform delivering 131 I-Hyp suppresses post-ablation residual lesions in colorectal cancer metastases via necrosis-targeted radiotherapy

doi: 10.1016/j.jpha.2025.101488

Figure Lengend Snippet: CD44 receptor expression and HA-PCL@( 131 I-Hyp) nanoparticles (HP-NPs) binding affinity in vitro . (A) Immunocytochemistry of CD44 expression in HT-29 and HCT-15 cells assessed by fluorescence microscopy with a fluorescein isothiocyanate (FITC) dye. (B) Flow cytometry analysis of CD44 surface expression in HT-29 and HCT-15 colorectal cancer cell lines. Cells were analyzed using unstained isotype controls for gating. CD44-positive cells represented 86.8% of the HT-29 population versus 0.65% of the HCT-15 population. (C) Intracellular distribution of fluorescence from hypericin (Hyp) at 0.5, 1, and 2 h posttreatment in HT-29 cells, HCT-15 cells, and HT-29 cells treated with free hyaluronic acid (HA). Red fluorescence: Hyp; blue fluorescence: Hoechst. (D, E) Flow cytometry (D) and quantitative analysis (E) of the fluorescence intensity of HT-29 and HCT-15 cells incubated with the same concentration of HP-NPs. ∗ P < 0.05, ns: no significant. HA-PCL: hyaluronan- b -poly(ε-caprolactone); DAPI: 4',6-diamidino-2-phenylindole.

Article Snippet: Anti-human/mouse CD44 conjugated with fluorescein isothiocyanate (FITC) (F1104401; Lianke Biotech Co., Ltd., Hangzhou, China) was added to the cells in the experimental groups at a 1:20 dilution in flow staining buffer, and the mixture was incubated at room temperature for 15 min in the dark.

Techniques: Expressing, Binding Assay, In Vitro, Immunocytochemistry, Fluorescence, Microscopy, Flow Cytometry, Incubation, Concentration Assay