Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: Material characterization of Au x Cu y Pd z @PSMA nanoparticles with different Pd ratios (0–20%): (a) UV–Visible spectra of as-prepared Au x Cu y Pd z nanoparticles and TEM images of (b) Au 72 Cu 28 , (c) Au 69 Cu 23 Pd 8 , (d) Au 57 Cu 16 Pd 27 , and (e) Au 36 Cu 5 Pd 59 nanocomposites. (f) AAS elemental analysis of Au x Cu y Pd z nanoparticles.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques:

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: (a) High-magnification TEM image of the Au 36 Cu 5 Pd 59 nanoparticles and corresponding EDS mapping images and (b) line-scan analysis of Au, Cu, and Pd. XPS spectra for the binding energies of (c) Cu 2p, (d) Au 4f, and (e) Pd 3d + Au 4d by the Au 36 Cu 5 Pd 59 nanoparticles. UV–visible records for the (f) catalytic reduction of 4-NP and (g) catalytic oxidation of TMB by the Au 72 Cu 28 , Au 69 Cu 23 Pd 8 , Au 57 Cu 16 Pd 27 , and Au 36 Cu 5 Pd 59 nanoparticles.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: Binding Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: Cytotoxicity and autophagy induction effects of Au x Cu y Pd z nanoparticles with different Pd ratios on bladder cancer cells. MTT assay with (a) Au x Cu y Pd z nanoparticles at 24 h and (b,c) different incubation times in T24 human bladder cancer cells. (d) Au 72 Cu 28 and Au 36 Cu 5 Pd 59 differentially induced autophagy in T24 human bladder cancer cells at different concentrations at 8 h, as determined via AO staining and flow cytometry analysis. The results were statistically analyzed, with three replicates shown in (e). (f) Confocal microscopy images of T24 cells after treatment with different concentrations of Au 72 Cu 28 or Au 36 Cu 5 Pd 59 for 24 h, followed by LC3 and Lamp-2 immunofluorescence staining. Scale bar = 50 μm. (g) Changes in the expression levels of the autophagy protein LC3 in T24 cells after being treated with 0.02 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 for 24 h, along with the quantification results. (h) Changes in the expression levels of the autophagy protein LC3 in T24 cells after being treated with different concentrations of the Au 36 Cu 5 Pd 59 micronanoshells for 24 h, along with the quantification results.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: MTT Assay, Incubation, Staining, Flow Cytometry, Confocal Microscopy, Immunofluorescence, Expressing

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: Cell death type analysis. (a) Apoptosis results of cancer cells treated with different concentrations of Au 72 Cu 28 and Au 36 Cu 5 Pd 59 NPs via flow cytometric analysis of T24 cells after 4 h of culture. (b) Statistical analysis of apoptosis ratios from triplicate experiments in (a). Fluorescence images of mitochondrial ROS in cancer cells treated with 0.01 and 0.05 mM (c) Au 72 Cu 28 and (d) Au 36 Cu 5 Pd 59 NPs in T24 cells after 1, 2, and 4 h of culture with different concentrations of Au x Cu y Pd z and the related quantitation results (in the below). Scale bar = 50 μm. (e) Mitophagy fluorescence images of T24 cells treated with different concentrations of Au x Cu y Pd z for 8 and 16 h of culture were obtained with a mitophagy detection kit and the related quantitation results. Scale bar = 50 μm.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: Fluorescence, Quantitation Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: (a) LPO activity in T24 cells treated with different concentrations of Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NPs after 4 and 16 h of culture. (b) Statistical analysis of LPO ratios from triplicate experiments in (a). n = 3. (c) Changes in the expression levels of GPX4 in T24 cells after coculture with indicated concentrations of Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NPs for 16 h, as determined by Western blotting. (d) The quantification results and (e) fluorescent images of intracellular labile iron level analysis in T24 cells treated with Au x Cu y Pd z NPs for 8 and 16 h by FerroOrange staining, Scale bar = 50 μm.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: Activity Assay, Expressing, Western Blot, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: (a) Cell uptake of Au x Cu y Pd z NPs was measured via dark-field microscopy images of T24 cells treated with 0.025 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NPs at different time points (0–24 h). Scale bar = 20 μm. (b) Normalized punctate signals within the cells from (a). (c) Total uptake of Au in T24 cells by atomic absorption spectroscopy during the first 6 h of incubation with Au x Cu y Pd z micronanoshells, followed by an additional 6 and 24 h after removing the NPs. (d) Cu metabolism-related gene expression analysis of T24 treated with Au x Cu y Pd z materials for 24 h via qPCR. (e) Schematic illustration showing that Pd-doped AuCu NPs enhanced cell uptake and exocytosis while they still induced ROS and autophagy, leading to cell death. The statistics are compared between Au 72 Cu 28 and Au 36 Cu 5 Pd 59 NPs at the same time points.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: Microscopy, Atomic Absorption Spectroscopy, Incubation, Gene Expression

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: Cytotoxicity results of (a) MTT of T24 cells treated with equal amounts of metal ions for 24 h. (b) MTT results of Au 36 Cu 5 Pd 59 micronanoshells combined with autophagy/Cu/ferroptosis inhibitors or activators. (c) Analysis of the intracellular labile iron level in Au x Cu y Pd z -treated T24 cells via FerroOrange staining under 20 μM CQ cotreatment for 16 h, and the fluorescence quantification results are presented in (d). Scale bar = 50 μm. (e) Changes in the expression levels of IDO1, PD-L1, and CD47 in T24 cells after treatment with indicated concentrations of Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP for 16 h, as determined by Western blotting. (f) The scheme illustrates that the Au x Cu y Pd z NPs enhanced cancer cell uptake and induced ROS, which led to subsequent lipid peroxidation (ferroptosis), sustained autophagy (mitophagy), and Cu metabolism alterations (upregulated CTR1/2, ATOX, and ATP7A/B) and ultimately caused cell death. The sustained autophagy additionally triggered the degradation of immune evasion proteins (PD-L1, CD47, IDO-1) via lysosomes and proteasomes, resulting in the reversal of the immunosuppressive tumor microenvironment (right seesaw). Using inhibitors or reagents for blocking uptake, autophagy or copper chelation can significantly rescue cells from death.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: Staining, Fluorescence, Expressing, Western Blot, Blocking Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Development of Au x Cu y Pd z Nanocomposites as Therapeutic Agents: Enhancing Cancer Treatment through Autophagy Modulation and Immune-Associated Effects

doi: 10.1021/acsami.5c20536

Figure Lengend Snippet: In vivo mechanism validation by orthotopic MB49 tumor-bearing mice. (a) Schematic illustration of treatment groups and experiment timelines for the mouse orthotopic bladder cancer model. Ultrasound imaging (b) and the normalized growth curve (c) of the tumor growth for the MB49 tumor-bearing mice received Au 36 Cu 5 Pd 59 micronanoshells and Au 86 Cu 14 nanoshells. The related survival rate and body weight curve are shown in (d,e), respectively. Scale bar = 2 mm in (b). (f) The major organ (heart, lung, liver, kidney, spleen, tumor) H&E staining images of Au 36 Cu 5 Pd 59 micronanoshells, Au 86 Cu 14 nanoshells, and particle-free groups. Scale bar = 100 μm. (g) The biodistribution of Au 36 Cu 5 Pd 59 micronanoshells at different time points (1 h, 1 day, 3 days, and 7 days). N.D. means less than 0.1 ppm. (h) The tumor IHC images of Au 36 Cu 5 Pd 59 micronanoshells, Au 86 Cu 14 nanoshells, and particle-free groups for autophagy, ferroptosis, and immune cell markers. Scale bar = 200 μm.

Article Snippet: T24 and MB49 cells cultured in 6-well dishes (2 × 10 5 per well) subjected to 0.01, 0.025, and 0.05 mM Au 72 Cu 28 or Au 36 Cu 5 Pd 59 NP treatments (w/o CQ and MG-132) for 8 and 16 h were harvested and lysed in lysis buffer (M-PER mammalian protein extraction reagent, Thermo Fisher Scientific, Inc.) for 30 min on ice and then centrifuged at 14,000 rpm for 20 min at 4 °C to remove precipitates.

Techniques: In Vivo, Biomarker Discovery, Imaging, Staining

Journal: International Journal of Pharmaceutics: X

Article Title: Gold-incorporated hyaluronic acid nanoparticles enhance ablative radiotherapy efficacy in lung cancer

doi: 10.1016/j.ijpx.2025.100480

Figure Lengend Snippet: Preparation and characteristics of Au-HA-functionalized SN38 NPs. (a) HA NPs are prepared by complexing SN38 with a human serum albumin (HSA)-based HA/PEI mixture. Simultaneously, Au NP-incorporated HA NPs (Au/HA NPs) are synthesized by integrating PEI-stabilized Au NPs into the HA/albumin complex. HA-functionalized SN38 NPs are prepared using a lyophilization–hydration method. (b) Transmission electron microscopy images of HA NPs and Au/HA NPs. Samples are negatively stained with 2 % uranyl acetate before imaging. Scale bar = 200 nm. (c) The average size and zeta potential of HA NPs and Au/HA NPs are analyzed using dynamic light scattering and aqueous electrophoresis. Each experimental group includes five samples ( n = 5). Data are expressed as the mean ± standard error of the mean. Statistical analyses are performed using the Mann–Whitney U test. (d) Full scan XPS spectrum of HA NPs and Au/HA NPs. (e) X-ray diffraction patterns of SN38, HA NP, and Au/HA NPs. (f) Storage stability of NPs as evaluated according to particle size (D) is determined by dissolving the re-lyophilized HA NPs and Au/HA NPs in 1 mL double-distilled water (ddH 2 O). Each experimental group include five samples ( n = 5). (g) Colloidal stability of NPs, as measured according to particle size (D), is assessed in Dulbecco's Modified Eagle Medium (Gibco) supplemented with 10 % ( v /v) fetal bovine serum to evaluate stability under physiologically relevant conditions. Each experimental group includes five samples (n = 5). (h) Representative and (i) quantitative cell surface CD44 expression posttreatment with SN38, HA NPs, and Au/HA NPs in A549, H226, and LLC cells as determined using flow cytometry. Each experimental group includes five samples (n = 5). Data are expressed as the mean ± standard error of the mean. Statistical significance is assessed using the one-way analysis of variance with Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: HA, hyaluronic acid; LLC, Lewis lung carcinoma; NP, nanoparticle; PEI, polyethyleneimine.

Article Snippet: Powder X-ray diffraction (XRD) patterns of SN38, HA NP, and Au/HA NP were recorded using a D2 PHASER X-ray diffractometer (Bruker Analytical X-Ray Solutions, Madison, WI, USA) equipped with a Cu Kα radiation source (λ = 1.5418 Å).

Techniques: Synthesized, Lyophilization, Transmission Assay, Electron Microscopy, Staining, Imaging, Zeta Potential Analyzer, Electrophoresis, MANN-WHITNEY, Modification, Expressing, Flow Cytometry

Journal: International Journal of Pharmaceutics: X

Article Title: Gold-incorporated hyaluronic acid nanoparticles enhance ablative radiotherapy efficacy in lung cancer

doi: 10.1016/j.ijpx.2025.100480

Figure Lengend Snippet: In vitro cytotoxicity and radiosensitization effects of Au-hyaluronic acid (HA)-functionalized SN38 NPs in A549 (human adenocarcinoma), H226 (human squamous cell carcinoma), and Lewis lung carcinoma (murine lung carcinoma) lung cancer cell lines. (a) In vitro cytotoxicity effect of SN38, HA NPs, and Au/HA NPs in A549, H226, and LLC cells. Cells are treated with different concentrations of SN38, HA NPs, and Au/HA NPs. Cell viability is determined using a Cell Counting Kit-8 (Sigma–Aldrich, Saint Louis, MO, USA) assay. Each experimental group includes six samples per concentration ( n = 6). (b) Response of lung cancer cells to radiation (0, 2, 4, 6, or 8 Gy) in the presence of SN38, HA NPs, and Au/HA NPs, as determined via colony formation assay. Each experimental group includes five samples (n = 5). (c) Quantification of γH2AX foci per cell, as a marker of DNA double-strand breaks after exposure to ionizing radiation, in A549, H226, and LLC cells treated with SN38, HA NPs, or Au/HA NPs and ablative radiotherapy (RT, 6 Gy). Each experimental group includes six samples (n = 6). (d) Representative immunofluorescence merged images of γ-H2AX foci shown in green (AlexaFluor488) and nuclei in blue (Hoechst 33342) in LLC cells treated with SN38, HA NPs, or Au/HA NPs and 6-Gy RT. (e) Analysis of cell cycle distribution. Each experimental group comprised five samples (n = 5). (f) Representative cell cycle distribution of A549, H226, and LLC cells treated with SN38, HA NPs, or Au/HA NPs and 6 Gy RT, as analyzed via flow cytometry. Data are expressed as the mean ± standard error of the mean. Statistical significance is assessed using the one-way analysis of variance with Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: HA, hyaluronic acid; LLC, Lewis lung carcinoma; NP, nanoparticles; RT, radiotherapy. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Powder X-ray diffraction (XRD) patterns of SN38, HA NP, and Au/HA NP were recorded using a D2 PHASER X-ray diffractometer (Bruker Analytical X-Ray Solutions, Madison, WI, USA) equipped with a Cu Kα radiation source (λ = 1.5418 Å).

Techniques: In Vitro, Cell Counting, Concentration Assay, Colony Assay, Marker, Immunofluorescence, Flow Cytometry

Journal: International Journal of Pharmaceutics: X

Article Title: Gold-incorporated hyaluronic acid nanoparticles enhance ablative radiotherapy efficacy in lung cancer

doi: 10.1016/j.ijpx.2025.100480

Figure Lengend Snippet: Radiation-induced ICD through ROS generation, CRT expression, and extracellular ATP release. ROS generation induces nuclear damage and triggers ICD. Intracellular ROS are detected by DCFH-DA staining. (a) Representative ROS generation and (b) quantitative intracellular ROS generation after treatment with SN38, HA-functionalized SN38 NPs, Au-HA NPs, or RT (6 Gy) in H226 and LLC cells, as determined using flow cytometry. Each experimental group includes three samples ( n = 3). (c) Representative CRT expression. (d) Quantification of CRT expression in lung cancer cells after RT combined with SN38, HA NPs, or Au/HA NPs, as determined using flow cytometry. Each experimental group includes three samples (n = 3). (e) Quantification of extracellular ATP release of lung cancer cells after RT treatment combined with SN38, HA NPs, or Au/HA NPs. Each experimental group includes three samples (n = 3). Data are expressed as the mean ± standard error of the mean. Statistical significance is assessed using the one-way analysis of variance with Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: CRT, calreticulin; HA, hyaluronic acid; ICD, immunogenic cell death; LLC, Lewis lung carcinoma; NPs, nanoparticles; ROS, reactive oxygen species; RT, radiotherapy.

Article Snippet: Powder X-ray diffraction (XRD) patterns of SN38, HA NP, and Au/HA NP were recorded using a D2 PHASER X-ray diffractometer (Bruker Analytical X-Ray Solutions, Madison, WI, USA) equipped with a Cu Kα radiation source (λ = 1.5418 Å).

Techniques: Expressing, Staining, Flow Cytometry

Journal: International Journal of Pharmaceutics: X

Article Title: Gold-incorporated hyaluronic acid nanoparticles enhance ablative radiotherapy efficacy in lung cancer

doi: 10.1016/j.ijpx.2025.100480

Figure Lengend Snippet: In vivo antitumor effect of Au/HA-functionalized SN38 NPs combined with RT. (a) Nude mice are subcutaneously implanted with human adenocarcinoma (A549) cells (2 × 10 6 ) on the flanks. Flank solid tumors (38 ± 14 mm 3 ) develop 14 days after inoculation. Mice are then randomized to treatment: irinotecan (20 mg/kg on day 0, administered intravenously), HA SN38 NPs (20 mg SN38/kg on day 0, administered intravenously), or Au-modified HA SN38 NPs (20 mg/kg SN38 on day 0, administered intravenously), with or without ablative RT (12 Gy/day for 2 days on days 0 and 1) to the flank tumor. Each experimental group involves five mice (n = 5). (b) Tumor images, (c) relative tumor growth curves, and (d) tumor volumes on day 56 post-RT of the subcutaneous A549 xenograft flank tumor model. (e) Synchronous LLC lung and flank tumor model. C57BL/6 mice undergo intrapulmonary injection and flank subcutaneous implantation with murine LLC cells. C57BL/6 mice first receive an intrapulmonary injection of 2 × 10 3 LLC cells for tumor development. After 3 days, mice are subcutaneously injected with 2 × 10 5 LLC cells in the flank. Flank solid tumors (36 ± 13 mm 3 ) develop 7 days after inoculation. Tumor-bearing mice are randomized into eight groups, as described above. The control group consisted of five mice (n = 5), whereas each treatment group consisted of six mice (n = 6). (f) Relative flank tumor growth curves and (g) Kaplan–Meier curves of overall survival for the synchronous LLC lung and flank tumor model mice. Mice are euthanized 3 weeks after intrapulmonary injection of LLC cells to assess both lung and flank tumor burdens, (h) flank tumor weights, and (i) lung weights as a proxy of intrapulmonary tumor burden and to obtain (j) lung images from the front and back. Data are expressed as the mean ± standard error of the mean. Statistical analyses are performed using the one-way analysis of variance with Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: HA, hyaluronic acid; i.v., intravenous; LLC, Lewis lung carcinoma; NP, nanoparticle; RT, radiotherapy; s.c., subcutaneous.

Article Snippet: Powder X-ray diffraction (XRD) patterns of SN38, HA NP, and Au/HA NP were recorded using a D2 PHASER X-ray diffractometer (Bruker Analytical X-Ray Solutions, Madison, WI, USA) equipped with a Cu Kα radiation source (λ = 1.5418 Å).

Techniques: In Vivo, Modification, Injection, Control

Journal: International Journal of Pharmaceutics: X

Article Title: Gold-incorporated hyaluronic acid nanoparticles enhance ablative radiotherapy efficacy in lung cancer

doi: 10.1016/j.ijpx.2025.100480

Figure Lengend Snippet: Quantitative flow cytometric analysis of subpopulations of tumor-infiltrating immune cell in the flank and lung tumor microenvironment. (a) C57BL/6 mice undergo subcutaneous implantation of murine LLC cells in the flanks and are randomized to treatment: irinotecan (20 mg/kg on day 0, administered intravenously), hyaluronic acid SN38 nanoparticles (HA NPs, 20 mg SN38/kg on day 0, administered intravenously), or Au-modified HA SN38 NPs (Au/HA NPs, 20 mg SN38/kg on day 0, administered intravenously), with or without ablative radiotherapy (12 Gy/day for 2 days on days 0 and 1) to the flank tumor. TIICs are isolated from the flank tumor microenvironment to obtain cell suspensions for surface staining. (b) Quantitative data of the percentage of tumor-infiltrating immune cells in the flank tumor environment, CD3 − CD49b + NK cells and CD3 + CD49b + NK T cells, CD4 + T cells, CD11c + dendritic cells, CD8 + T cells, and CD4 + FoxP3 + regulatory T cells and the ratio of CD8 + T cells and regulatory T cells are shown. (c) C57BL/6 mice receive intrapulmonary injections of murine LLC cells, undergo subcutaneous implantation with murine LLC cells in the flanks, and are randomized to treatment as described above. Tumor-infiltrating immune cells are isolated from the non-irradiated lung tumor microenvironment to obtain cell suspensions for surface staining. (d) Quantitative data of the percentage of tumor-infiltrating immune cells in the non-irradiated lung tumor microenvironment, CD3 − CD49b + NK cells and CD3 + CD49b + NK T cells, CD11c + dendritic cells, and CD8 + T cells are shown. (e) Representative flow cytometric analysis, CD3 − CD49b + NK cells and CD3 + CD49b + NK T cells, CD11c + dendritic cells, and CD8 + T cells in the flank tumor microenvironment (left panel) and the non-irradiated lung tumor microenvironment (right panel) are shown. The control group consisted of five mice (n = 5), whereas each treatment group consisted of six mice (n = 6). Data are expressed as the mean ± standard error of the mean. Statistical analyses are performed using the one-way analysis of variance with Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: HA, hyaluronic acid; i.v., intravenous; LLC, Lewis lung carcinoma; NK, natural killer cell; NKT, natural killer T cell; NP, nanoparticle; RT, radiotherapy; s.c., subcutaneous.

Article Snippet: Powder X-ray diffraction (XRD) patterns of SN38, HA NP, and Au/HA NP were recorded using a D2 PHASER X-ray diffractometer (Bruker Analytical X-Ray Solutions, Madison, WI, USA) equipped with a Cu Kα radiation source (λ = 1.5418 Å).

Techniques: Modification, Isolation, Staining, Irradiation, Control

Journal: International Journal of Pharmaceutics: X

Article Title: Gold-incorporated hyaluronic acid nanoparticles enhance ablative radiotherapy efficacy in lung cancer

doi: 10.1016/j.ijpx.2025.100480

Figure Lengend Snippet: In vivo biodistribution of Au-HA-functionalized SN38 nanoparticles and CD44 and CRT expression in flank tumors. (a) Distribution of HA NPs and Au/HA NPs in the main organs and LLC tumors at 4 and 24 h. (b) Representative immunohistochemical images of CD44 expression in A549 (human adenocarcinoma) flank tumor tissue treated with HA NPs or Au/HA NPs. Scale bar = 60 μm. (c) Quantitative results for CD44 expression. The data represent the percentage of CD44-stained area within the observed area. Each experimental group involves three mice (n = 3). (d) Quantitative results for CRT expression. The data represent the percentage of CRT-stained areas within the observed area. Each experimental group involves three mice (n = 3). (e) Representative immunohistochemical images of CRT expression in LLC flank tumor tissue treated with radiotherapy alone or in combination with SN38, HA NPs, or Au/HA NPs. Scale bar = 60 μm. Data are expressed as the mean ± standard error of the mean. Statistical analyses are performed using the one-way analysis of variance with Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: CRT, calreticulin; HA, hyaluronic acid; LLC, Lewis lung carcinoma; NP, nanoparticle; RT, radiotherapy.

Article Snippet: Powder X-ray diffraction (XRD) patterns of SN38, HA NP, and Au/HA NP were recorded using a D2 PHASER X-ray diffractometer (Bruker Analytical X-Ray Solutions, Madison, WI, USA) equipped with a Cu Kα radiation source (λ = 1.5418 Å).

Techniques: In Vivo, Expressing, Immunohistochemical staining, Staining