Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: Schematic illustration of photoresponsive IR783/BC NP and its therapeutic effect upon light irradiation. (a) Self‐assembly of IR783/BC NP and its dissociation upon light irradiation. (b) Cellular uptake of IR783/BC NP by a HCT116 cell and light‐triggered drug release for cancer therapy

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: Irradiation

Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: Preparation and characterization of IR783/BC NPs. (a) Nanoparticle preparation by the flash nanoprecipitation method. (b) Representative images of free IR783, free BC prodrug, and IR783/BC NPs in PBS. (c) Representative size and (d) zeta potential distribution of IR783/BC NPs. (e) Stability test of IR783/BC NPs at 37°C in PBS for 48 h. (f) Size distributions of IR783/BC NPs at 0, 24, and 48 h

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: Zeta Potential Analyzer

Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: Light‐triggered photocleavage, nanoparticle dissociation and drug release. (a) TEM image of IR783/BC NPs without light irradiation. (b) TEM image of IR783/BC NPs after light irradiation for 5 min. (c) Size distributions of IR783/BC NPs under light irradiation for 0, 3, and 5 min. (d) HPLC analysis of IR783/BC NPs upon light irradiation. (e) Quantitative analysis of IR783/BC degradation and Cb release ( n = 3). Light irradiation: 530 nm, 50 mW/cm 2

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: Irradiation

Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: Spectrum study and photochemistry mechanisms. (a) UV‐Vis absorption spectra of IR783/BC NPs upon 530 nm light irradiation. Fluorescence spectra of (b) IR783/BC NPs and (c) free IR783 upon 530 nm light irradiation (Ex. 783 nm). (d) Photocleavage process of BC prodrug. (e) ROS‐triggered decomposition of IR783. The LC‐MS characterization of intermediates and products ①–⑧ is showed in Figure . Light irradiation: 530 nm, 50 mW/cm 2

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: Irradiation, Fluorescence, Liquid Chromatography with Mass Spectroscopy

Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: In vitro cellular uptake, cytotoxicity and 1 O 2 generation. (a) CLSM images of HCT116 cells incubated with free IR783 and IR783/BC NPs for 6 h. Scale bar: 20 μm. Blue channel: ex. 405 nm; Red channel: ex. 639 nm, with a 650 nm optical filter. (b) In vitro cytotoxicity test of free BC prodrug and IR783/BC NPs with/without light irradiation by MTT assay. * p < 0.05, ** p < 0.01. (c) Calcein‐AM/PI staining of HCT116 cells after treatment with free BC, free IR783, IR783/BC NPs with/without light irradiation. Scale bar: 100 μm. (d) Fluorescent change of IR783/BC NPs solution containing SOSG as 1 O 2 sensor. (e) The 1 O 2 production of IR783, free BC, and IR783/BC NPs in the presence of light. (f) CLSM images of intracellular 1 O 2 in HCT116 cells incubated with IR783/BC NPs at different concentrations with/without light irradiation. DCFH‐DA was used as the indicator. Scale bar: 10 μm. Light irradiation: 530 nm, 50 mW/cm 2 , 5 min (except d and e)

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: In Vitro, Incubation, Irradiation, MTT Assay, Staining

Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: In vivo biodistribution and in‐situ monitoring upon light irradiation. (a) Schematic illustration of the treatment process in HCT116 tumor‐bearing mouse. (b) Representative IVIS fluorescent images of the mice post‐injection of free IR783 or IR783/BC NPs within 24 h ( n = 3) (Ex. 780 nm). (c) Quantitative analysis of the biodistribution determined by IVIS in major organs and tumors. Tu, He, Lu, Sp, Li, and Ki represent tumor, heart, lung, spleen, liver, and kidney, respectively. ** p < 0.01. (d) Quantitative analysis of the penetration efficiency of 530 nm green light through mouse skin samples ( n = 3) within 100 s. (e) Quantitative analysis of fluorescence intensity in the tumor regions exposed to light irradiation for various durations. (f) In‐situ fluorescence imaging of the mice at 24 h post‐injection of IR783/BC NPs with light irradiation at the tumor sites for various durations ( n = 3). The tumor sites are marked by white circles. Light irradiation: 530 nm, 100 mW/cm 2

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: In Vivo, In Situ, Irradiation, Injection, Fluorescence, Imaging

Journal: Bioengineering & Translational Medicine

Article Title: Photoresponsive prodrug‐dye nanoassembly for in‐situ monitorable cancer therapy

doi: 10.1002/btm2.10311

Figure Lengend Snippet: Antitumor efficacy in HCT116 tumor model. (a) Schematic illustrating treatment schedule for inhibiting subcutaneous tumor growth. (b) Tumor volume changes after treatments with PBS, IR783, Cb, IR783/BC NPs, and IR783/BC NPs with light irradiation, separately ( n = 4). ** p < 0.01. (c) Photos of the mice with subcutaneous tumors, (d) tumor weight, (e) photos of the tumors at Day 14 after different treatments as indicated. (f) Changes in body weight of the mice during the treatments. Light irradiation: 530 nm, 100 mW/cm 2 , 10 min

Article Snippet: Indocyanine IR783 dye was purchased from Tokyo Chemical Industry (TCI) Co., Ltd. Chlorambucil (Cb) was purchased from J&K Scientific.

Techniques: Irradiation

Journal:

Article Title: Near Infrared Heptamethine Cyanine Dye-Mediated Cancer Imaging *

doi: 10.1158/1078-0432.CCR-10-0059

Figure Lengend Snippet: Active uptake of hepatamethine cyanine dyes by human cancer cells but not normal cells in culture. (A), The chemical structures of two hepatamethine cyanine dyes, IR-783 and MHI-148. (B), Normal human cells including bone marrow stromal cells (HS-27A), normal prostate epithelial cells (NPE), normal prostate stromal fibroblasts (NPF), vascular endothelial cells (HUVEC-CS) and human embryonic kidney cells (HEK293) showed very low uptake of these dyes in culture. (C), Human cancer cell lines including prostate (C4-2, PC-3, ARCaPM), breast (MCF-7), cervical (HeLa), lung (H358), liver (HepG2), pancreatic (MIA PaCa-2) and renal (SN12C) cancer cells, as well as a human leukemia cell line (K562), showed significant uptake of IR-783 dye under similar staining and imaging conditions. Results are shown with images obtained from cells stained with DAPI of cell nuclei, the heptamethine cyanine IR-783 stain (NIR), and a merger of the two images (Merge). All the images were acquired at 630 × magnification.

Article Snippet: To assess tissue distribution of these dyes, athymic mice without tumor implantation were sacrificed at 0, 6 and 80 hrs (N=3 each) after i.v injection of IR-783 dye at a dose of 10 nmol/20 g. Dissected organs were subjected to NIR imaging by a Kodak Imaging Station 4000 MM.

Techniques: Staining, Imaging

Journal:

Article Title: Near Infrared Heptamethine Cyanine Dye-Mediated Cancer Imaging *

doi: 10.1158/1078-0432.CCR-10-0059

Figure Lengend Snippet: Kinetics and subcellular localization of the NIR dyes. (A), Confocal imaging shows significant uptake of IR-783 dye in ARCaPM cells but not in normal human prostate epithelial P69 cells at 630× magnification. (B), Histogram shows differential and time-dependent uptake of IR-783 by human prostate cancer ARCaPM cells and P69 cells. (C), Uptake of the IR-783 dye (20 μM) by ARCaPM cells can be abrogated by 250 μM BSP. (D), Subcellular co-localization of the NIR hepatamethine cyanine dyes with lysosomes (Lyso) and mitochondrial (Mito) tracking dyes. ARCaPM cells that were stained with IR-783 were stained with a lysosome-specific dye, Lyso Tracking Green DND-26, and a mitochondria-specific dye, Mito Tracker Orange CMTMROS (630×). Fluorescence imaging indicates that a large portion of the IR-783 was co-localized with these subcellular organelles.

Article Snippet: To assess tissue distribution of these dyes, athymic mice without tumor implantation were sacrificed at 0, 6 and 80 hrs (N=3 each) after i.v injection of IR-783 dye at a dose of 10 nmol/20 g. Dissected organs were subjected to NIR imaging by a Kodak Imaging Station 4000 MM.

Techniques: Imaging, Staining, Fluorescence

Journal:

Article Title: Near Infrared Heptamethine Cyanine Dye-Mediated Cancer Imaging *

doi: 10.1158/1078-0432.CCR-10-0059

Figure Lengend Snippet: Preferential uptake and retention of the hepatamethine cyanine dyes in human tumor xenografts. Mice bearing human prostate (ARCaPM, orthotopic prostate tumor, p.o), bladder (T24, subcutaneous, s.c), pancreatic (MIA PaCa-2, subcutaneous), and renal (SN12C, intraosseous to tibia, i.o) tumors were injected i.p with IR-783 at a dose of 10 nmol/20g. NIR imaging was performed 24 hrs later. Each mouse was subjected to fluorescence imaging (NIR) and X-ray imaging (X-ray) using the Kodak Imaging Station Imaging System, and the two images were superimposed (Merge) for tumor localization. After imaging, tissues with specific fluorescence signals were dissected, fixed in 10 % formaldehyde, and subjected to histopathologic analysis by H/E staining (200×). In mice bearing subcutaneous tumors, we detected both tumors based on fluorescence imaging (see arrows).

Article Snippet: To assess tissue distribution of these dyes, athymic mice without tumor implantation were sacrificed at 0, 6 and 80 hrs (N=3 each) after i.v injection of IR-783 dye at a dose of 10 nmol/20 g. Dissected organs were subjected to NIR imaging by a Kodak Imaging Station 4000 MM.

Techniques: Injection, Imaging, Fluorescence, Staining

Journal:

Article Title: Near Infrared Heptamethine Cyanine Dye-Mediated Cancer Imaging *

doi: 10.1158/1078-0432.CCR-10-0059

Figure Lengend Snippet: Detection of tumor metastasis in mice and spontaneous tumors in transgenic animals. (A) Confirmation of the presence of bone metastatic prostate tumors in mice by NIR imaging after IR-783 i.p injection at a dose of 10 nmol/20g. (a), The ARCaPM human prostate cancer cell line was stably transfected with AsRed2 RFP. The clone being used in this study exhibited typical ARCaPM cell morphology (bright field, 100×) and could emit intense red fluorescence. (b), Cells from this clone were inoculated orthotopically to athymic mice to produce both localized prostate tumor (thick arrow) and bone metastatic tumor (thin arrow), which were detected by IR-783 fluorescence imaging of the whole animal (left) and of the dissected skeletal bone (right). (c), To confirm the detection of metastasis, marrow cells from the affected tibia/femur were cultured and isolated cancer cells were found to express RFP. (d), ARCaPM cells in the metastatic tibial/femur tumor could also be seen in formaldehyde fixed sections, either by conventional H/E stain or directly by red florescence imaging. These analyses unanimously confirmed that the signals attained in IR-783 imaging reflect metastases of the orthotopic ARCaPM tumor.

Article Snippet: To assess tissue distribution of these dyes, athymic mice without tumor implantation were sacrificed at 0, 6 and 80 hrs (N=3 each) after i.v injection of IR-783 dye at a dose of 10 nmol/20 g. Dissected organs were subjected to NIR imaging by a Kodak Imaging Station 4000 MM.

Techniques: Transgenic Assay, Imaging, Injection, Stable Transfection, Transfection, Fluorescence, Cell Culture, Isolation, Staining

Journal:

Article Title: Near Infrared Heptamethine Cyanine Dye-Mediated Cancer Imaging *

doi: 10.1158/1078-0432.CCR-10-0059

Figure Lengend Snippet: Distribution of heptamethine cyanine dye IR-783 and its metabolites in tissues; time-course and concentration-dependent studies in normal and tumor-bearing mice. (A) Normal organs dissected at 0, 6 and 80 hrs after IR-783 iv injection at a dose of 10 nmol/20g were subjected to NIR dye imaging with a Kodak Imaging Station 4000 MM (see Methods). Note that at 80 hrs, IR-783 was completely cleared from all vital organs examined. (B) A representative mouse bearing orthotopic ARCaPM human prostate tumor was imaged after IR-783 10 nmol/20g iv injection at 0.5, 24, 48, 72 and 96 hrs. Note dye uptake and retention seen in an ARCaPM orthotopic tumor. (C) A representative mouse bearing a subcutaneous ARCaPM tumor subjected to NIR imaging after IR-783 i.p injection at a dose of 10 nmol/20g. The left panel shows the retention of IR-783 in the tumor 24 hrs after dye administration in whole body in vivo imaging. The right panel shows the ex vivo imaging of surgically dissected tissues which confirmed the uptake and retention of IR-783 in a surgically dissected ARCaPM tumor. Top row of this panel from left: liver, lung, and heart. Bottom row of this panel from left: spleen, kidneys, and tumor. Tumor tissue displayed strong signals in both in vivo and ex vivo imaging. (D) A standard curve was constructed based on the fluorescence emission intensity of IR-783 at 820 nm (see Methods) with the dye added to a PBS solution at concentrations of 0.5, 1, 2, 4, 8, 16 and 32 μM. The correlation coefficient between the fluorescence emission intensity and concentration of IR-783 was estimated to be r=0.9991 (see left panel). The apparent dye concentration (μg/g) in organs and tumor was calculated based on the standard curve established above (see right panel). The apparent dye concentration is defined here by the light emission intensity at 820 nm, which could include the parental IR-783 and its metabolites. Data are expressed as average ± SEM of 3 determinations.

Article Snippet: To assess tissue distribution of these dyes, athymic mice without tumor implantation were sacrificed at 0, 6 and 80 hrs (N=3 each) after i.v injection of IR-783 dye at a dose of 10 nmol/20 g. Dissected organs were subjected to NIR imaging by a Kodak Imaging Station 4000 MM.

Techniques: Concentration Assay, IV Injection, Imaging, Injection, In Vivo Imaging, Ex Vivo, In Vivo, Construct, Fluorescence

Journal:

Article Title: Near Infrared Heptamethine Cyanine Dye-Mediated Cancer Imaging *

doi: 10.1158/1078-0432.CCR-10-0059

Figure Lengend Snippet: Detection of human prostate cancer cells in human blood. (A), ARCaPM cells mixed with human blood were incubated with IR-783 and the particulate fractions containing normal healthy mononuclear cells and cancer cells were isolated using gradient centrifugation. The cells were resuspended in PBS for acquisition of fluorescent images under a confocal microscope. Significant uptake and retention of the dye could be detected in ARCaPM cells in a fluorescent field (white arrow), while mononuclear cells hardly showed any signals (black arrow). (B), To determine the sensitivity of this novel method for tumor cell detection, known numbers of ARCaPM cells (10 to 1,000 cells) were added to 1 ml of whole blood. Following gradient centrifugation, washing and re-suspension, positive fluorescent cancer cells were counted. Results presented in the histograph represent three separate experiments (n=3) with data expressed as mean +/-SEM.

Article Snippet: To assess tissue distribution of these dyes, athymic mice without tumor implantation were sacrificed at 0, 6 and 80 hrs (N=3 each) after i.v injection of IR-783 dye at a dose of 10 nmol/20 g. Dissected organs were subjected to NIR imaging by a Kodak Imaging Station 4000 MM.

Techniques: Incubation, Isolation, Gradient Centrifugation, Microscopy

Journal: ACS Omega

Article Title: Theranostic Nanoprobes Assisted NIR-II Fluorescence Imaging for Efficient Angiography and Tumor Therapy

doi: 10.1021/acsomega.5c01553

Figure Lengend Snippet: (a) Cytotoxicity of HeLa cells incubated with free IR820, IR-T, and IR-DT at different concentrations (50, 100, 150, and 200 μg/mL) for 4 h. (b) NIR confocal laser scanning microscopic images of HeLa cells incubated with IR-DT nanoprobes. Scale bar = 50 μm. Excitation for the nanoprobes was 640 nm, excitation for HOCHEST was 405 nm. (c) Flow cytometric analysis of HeLa cells incubated with various concentrations (50, 100, 150, and 200 μg/mL) of IR783, IR-T nanoprobes, and IR-DT nanoprobes in aqueous dispersion, respectively.

Article Snippet: For metabolic imaging, IR783 and IR-DT nanoprobes (200 μL, 2.7 mg/kg) were intravenously injected into mice and imaged at different time points (2, 4, 8, 24, 48, and 72 h) by using the NIR-II imaging system (MARS-Pathfinder) under 808 nm laser irradiation.

Techniques: Incubation, Dispersion

Journal: ACS Omega

Article Title: Theranostic Nanoprobes Assisted NIR-II Fluorescence Imaging for Efficient Angiography and Tumor Therapy

doi: 10.1021/acsomega.5c01553

Figure Lengend Snippet: (a) In vivo NIR-II fluorescence imaging of IR-DT nanoprobes and IR783-treated nude mice at different time points (2 h, 4 h, 8 h, 24 h, 48 h, and 72 h). (b) Corresponding fluorescence intensity analysis at different time points in (a). (c) Corresponding quantitative fluorescence analysis of main organs in (d).

Article Snippet: For metabolic imaging, IR783 and IR-DT nanoprobes (200 μL, 2.7 mg/kg) were intravenously injected into mice and imaged at different time points (2, 4, 8, 24, 48, and 72 h) by using the NIR-II imaging system (MARS-Pathfinder) under 808 nm laser irradiation.

Techniques: In Vivo, Fluorescence, Imaging