Journal: Pharmaceutics

Article Title: Design and Evaluation of Autophagy-Inducing Particles for the Treatment of Abnormal Lipid Accumulation

doi: 10.3390/pharmaceutics14071379

Figure Lengend Snippet: In vivo and organ DiR fluorescence imaging of genetically obese ob/ob mice following a single IV injection (120 µL) of soluble fluorophore or fluorescent particles (0.25% w/v ). ( a ) Autophagy-inducing fluorescent particles (NP DiR T-B) and plain fluorescent particles (NP DiR) accumulate in the liver over the course of 72 h. Images are representative of scans from 3–4 mice per group. ( b ) Quantification of whole-body fluorescence up to 72 h post a single IV injection. The percentage values reported are calculated from the 5 min timepoint, when the maximal fluorescence was observed, and are reported as mean ± s.e.m. Results were analyzed using one-way ANOVA followed by Sidak’s multiple comparison test, which revealed no significant difference between the NP DiR and the NP DiR T-B group for all timepoints. ( c , d ) Organ biodistribution of fluorescent NPs 24 h after a single intravenous (IV) injection in ob/ob mice. n = 3–4 mice per group. Ex vivo DiR fluorescence imaging on main organs (liver, spleen, pancreas and kidneys) ( c ) and quantification of DiR fluorescence ( d ). The percentage values reported are calculated from total organ fluorescence and presented as mean ± s.e.m ( d ).

Article Snippet: Mice were immediately injected in the retro-orbital vein with 120 µL of either free fluorophore (Xenolight DiR-Fluorescent Dye, PerkinElmer, Waltham, MA, USA, 125964), fluorescent nanoparticles (NP DiR, Near Infra-Red Fluorescent i-Particles ® , Adjuvatis, France) or fluorescent Tat-Beclin nanoparticles (NP DiR T-B) of 0.25% w/v solid content.

Techniques: In Vivo, Fluorescence, Imaging, IV Injection, Ex Vivo

Journal: Theranostics

Article Title: Reversal of Vascular Calcification and Aneurysms in a Rat Model Using Dual Targeted Therapy with EDTA- and PGG-Loaded Nanoparticles

doi: 10.7150/thno.16547

Figure Lengend Snippet: Schematic representation of the experiment. The experiment consisted of five groups (n=6 animals per group) with different NP treatments viz. DIR NPs, Blank NPs, EDTA+PGG NPs, BB-94 NPs and PGG NPs. Calcium chloride injury to the aorta was done on Day 0 following which the animals were allowed to develop aneurysm for 4 weeks. After 4 weeks, they were treated with NPs and euthanized at various chosen time points as shown in the legend. DIR NPs were administered once after 4 weeks, Blank and PGG NPs were injected twice biweekly after 6 and 8 weeks, and BB-94 NPs were injected one weekly starting at 6 weeks and through 9 weeks after injury. The EDTA+PGG NPs group was given two EDTA NP injections every week starting at 4 weeks and through 5 weeks after injury, immediately followed by biweekly PGG NP injections through 9 weeks after injury. All rats were euthanized at week 12.

Article Snippet: Briefly, fluorescent infra-red dye 1, 1-dioctadecyl-3, 3, 3, 3-tetramethylindotricarbocyanine iodide (DIR)-loaded NPs(DIR-NPs) were obtained by dissolving 250 mg of BSA (Seracare, MA) in 4 mL of deionized water.

Techniques: Injection

Journal: Theranostics

Article Title: Reversal of Vascular Calcification and Aneurysms in a Rat Model Using Dual Targeted Therapy with EDTA- and PGG-Loaded Nanoparticles

doi: 10.7150/thno.16547

Figure Lengend Snippet: In situ zymography for MMPs (A1), alizarin red staining for calcium (A2) and VVG staining for elastin (fibers stained black) (A3) for aortic samples 30 days after injury. In situ zymography confirmed the high activity of MMPs (-9 and-2). Alizarin red confirmed calcification in the media and adventitia. VVG showed the elastin damage. (B) NP accumulation after intravenous injection of EL-NP-DIR, 30 days after injury at the site of elastin damage (squares show sections of the abdominal aorta where CaCl 2 was applied); image taken 24 hrs after injection. (C) Organ distribution of fluorescent NPs 24 hrs after injection of EL-NP-DIR. (D) Cross-section of abdominal aorta showing NPs (EL-NP-DIR) targeting from the adventitial side and accumulating more in the degraded elastic lamina; DIR fluorescence (purple) and elastin (green autofluorescence). * indicates lumen.

Article Snippet: Briefly, fluorescent infra-red dye 1, 1-dioctadecyl-3, 3, 3, 3-tetramethylindotricarbocyanine iodide (DIR)-loaded NPs(DIR-NPs) were obtained by dissolving 250 mg of BSA (Seracare, MA) in 4 mL of deionized water.

Techniques: In Situ, Zymography, Staining, Activity Assay, Injection, Fluorescence

Journal: Science advances

Article Title: Endogenous/exogenous dual-responsive nanozyme for photothermally enhanced ferroptosis-immune reciprocal synergistic tumor therapy.

doi: 10.1126/sciadv.adq3870

Figure Lengend Snippet: Fig. 4. Cellular uptake and in vitro therapeutic effect. (A) Fluorescence images of 4T1 cells incubated with RhB-MCMSFT for various time intervals. h, hours. (B) Fluores- cence images of caspase-3 (Cas-3) immunofluorescence staining and LPO staining in 4T1 cells after incubation with PBS or MCMSFT. (C) The cell viability of 4T1 cells treated with MCMSFT in the presence of DFO (100 μM), Fer-1(10 μM), or DEVD (50 μM). (D) The cell viability and (E) live/dead cell staining images of 4T1 cells treated with different formulations with or without laser irradiation. The data are presented as the means ± SD, n = 3, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. NS, no significance difference. AM, acetoxymethyl ester.

Article Snippet: Near- infrared fluorescence dye DiR was purchased from Abbkine Scientific (Wuhan, China).

Techniques: In Vitro, Fluorescence, Incubation, Immunofluorescence, Staining, Irradiation

Journal: Science advances

Article Title: Endogenous/exogenous dual-responsive nanozyme for photothermally enhanced ferroptosis-immune reciprocal synergistic tumor therapy.

doi: 10.1126/sciadv.adq3870

Figure Lengend Snippet: Fig. 5. Potential mechanism of hybrid multiple cell deaths induction. (A) The proposed molecular mechanisms of MCMSFT-induced hybrid cell death including apop- tosis, ferroptosis, and necroptosis. (B) Representative Western blot images of GPX4, ferritin, Cas-3, and PD-L1 expression measurement after different treatments. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. (C) Bio-TEM images of 4T1 cells treated with PBS or MCMSFT. (D to G) Fluorescence staining images of intracellular Fe2+, ROS, GSH, and LPO detection in 4T1 cells after different treatments.

Article Snippet: Near- infrared fluorescence dye DiR was purchased from Abbkine Scientific (Wuhan, China).

Techniques: Western Blot, Expressing, Fluorescence, Staining

Journal: Science advances

Article Title: Endogenous/exogenous dual-responsive nanozyme for photothermally enhanced ferroptosis-immune reciprocal synergistic tumor therapy.

doi: 10.1126/sciadv.adq3870

Figure Lengend Snippet: Fig. 7. In vivo live imaging and therapeutic efficacy. (A) Fluorescence images of 4T1 tumor–bearing mice at different time points after intravenous administration of free DiR or DiR-CMSFT. (B) Time-dependent photothermal images of 4T1 tumor–bearing mice after intravenous administration of saline or MCMSFT with laser irradiation. (C) Schematic illustration of 4T1 subcutaneous tumor–bearing mouse model establishment and treatment schedule. (D) Photograph of tumors from mice after various treatments. (E) Representative hematoxylin and eosin (H&E), terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL), and Ki67 staining images of tumor sections. Scale bars, 100 μm. (F) Representative LPO, CD8, and Cas-3 staining images of tumor sections. Scale bars, 100 μm.

Article Snippet: Near- infrared fluorescence dye DiR was purchased from Abbkine Scientific (Wuhan, China).

Techniques: In Vivo, Imaging, Drug discovery, Fluorescence, Saline, Irradiation, End Labeling, TUNEL Assay, Staining

Journal: Science advances

Article Title: Endogenous/exogenous dual-responsive nanozyme for photothermally enhanced ferroptosis-immune reciprocal synergistic tumor therapy.

doi: 10.1126/sciadv.adq3870

Figure Lengend Snippet: Fig. 8. Ferroptosis-immune reciprocal effect and immune antitumor activity. (A) Representative fluorescence and immunofluorescence staining images of ROS, GPX4, and ferritin. Scale bars, 100 μm. (B) Representative immunofluorescence staining images of CRT and PD-L1. Scale bars, 100 μm. (C) ELISA measurement of IFN-γ levels in serum collected from mice after various treatments. (D) Representative immunofluorescence staining images of SLC7A11 and SLC3A2. Scale bars, 100 μm. (E) Representative flow cytometry plots and (F) quantitative analysis of mature DCs (CD11C+CD80+CD86+) in tumor-draining lymph nodes (LNs). (G to L) Representative flow cytometry plots and quantitative analysis of cytotoxic T lymphocytes (CD3+CD8+) and helper T lymphocytes (CD3+CD4+) in the spleen and tumor. The data are pre- sented as the means ± SD, n = 3, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Near- infrared fluorescence dye DiR was purchased from Abbkine Scientific (Wuhan, China).

Techniques: Activity Assay, Fluorescence, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Science advances

Article Title: Endogenous/exogenous dual-responsive nanozyme for photothermally enhanced ferroptosis-immune reciprocal synergistic tumor therapy.

doi: 10.1126/sciadv.adq3870

Figure Lengend Snippet: Fig. 9. Suppression of lung metastasis and rechallenge tumor. (A) Schematic illustration of lung metastasis tumor–bearing mouse model establishment and treatment schedule. (B and C) Photographs of Bouin’s solution–stained lung tissues and quantitative analysis of metastasis nodules with different treatments. (D) H&E and immuno- fluorescence staining images of CD8 and Ki67 in the lung sections with different treatments. (E) Schematic illustration of rechallenge tumor–bearing mouse model estab- lishment and treatment schedule. (F) Observation of primary tumors and rechallenge tumors growth with different treatments. (G) Quantitative analysis of the rechallenge tumor growth of mice with different treatments. (H) Immunofluorescence staining images of CD8 and Ki67 in the tumor sections with different treatments. (I) Survival curve of mice with different treatments. The data are presented as the means ± SD, n = 5, **P < 0.01 and ***P < 0.001.

Article Snippet: Near- infrared fluorescence dye DiR was purchased from Abbkine Scientific (Wuhan, China).

Techniques: Staining, Fluorescence, Immunofluorescence