Journal: Nanoscale horizons

Article Title: Nanoassemblies of heptamethine cyanine dye-initiated poly(amino acid) enhance ROS generation for effective antitumour phototherapy.

doi: 10.1039/d3nh00584d

Figure Lengend Snippet: Scheme 1 Schematic illustration of material synthesis and the formation of a polymeric nanoassembly. (A) Synthesis route of the materials. IR783 was firstly reacted with cysteamine to introduce the active thiol into the molecule. The IR783-cysteamine derivative then initiated the N-carboxyanhydride (NCA)- mediated ROP reaction to generate the polymer (IR783-pBLA). (B) The IR783- pBLA polymers self-assemble to form IR-PA nanoparticles in an aqueous environment by hydrophobic interactions, which are further stabilized by the negative surface charge and the helical structure of the pBLA block.

Article Snippet: IR783 (sodium salt), triethylamine (TEA), ethyl acetate, and 1,3- diphenylisobenzofuran (DPBF) were purchased from Tokyo Chemical Industry Co., Ltd (Tokyo, Japan).

Techniques: Introduce, Polymer, Blocking Assay

Journal: Nanoscale horizons

Article Title: Nanoassemblies of heptamethine cyanine dye-initiated poly(amino acid) enhance ROS generation for effective antitumour phototherapy.

doi: 10.1039/d3nh00584d

Figure Lengend Snippet: Fig. 1 Characterization of IR-PA. (A) Thermodynamically optimal conformation of the IR783-derivative materials. (B) Representative histogram showing the hydrodynamic size distribution of IR-PA in aqueous solution. (C) z-Potential of IR-PA in aqueous buffer with different pH. Data shown as mean S.D., n = 3. (D) Representative fluorescence emission spectra of IR-PA in different solvents. The emission intensities are normalized to the maximal emission intensity of IR-PA in DMSO. (E) Representative fluorescence image of IR-PA samples dispersed in different solvents. (Excitation filter: 640 nm, emission filter: 780 nm). (F) Representative absorbance spectra of 1,3-diphenylisobenzofuran (DBPF) incubated with IR-PA after irradiation with a 633 nm laser for different times. (G) ROS generation efficiency from IR783-cysteamine or IR-PA reflected by the absorbance intensity of DBPF at 430 nm after different treatments. Data shown as mean S.D., n = 3. (H) Absorbance spectra of IR783-cysteamine (black lines) and IR-PA (red lines) before (solid lines) and after (dashed lines) 30 min irradiation with a 633 nm laser. (I) Relative absorbance at 660 nm of IR783-cysteamine or IR-PA after irradiation with a 633 nm laser for different times. Data shown as mean S.D., n = 3.

Article Snippet: IR783 (sodium salt), triethylamine (TEA), ethyl acetate, and 1,3- diphenylisobenzofuran (DPBF) were purchased from Tokyo Chemical Industry Co., Ltd (Tokyo, Japan).

Techniques: Fluorescence, Incubation, Irradiation

Journal: Nanoscale horizons

Article Title: Nanoassemblies of heptamethine cyanine dye-initiated poly(amino acid) enhance ROS generation for effective antitumour phototherapy.

doi: 10.1039/d3nh00584d

Figure Lengend Snippet: Fig. 2 In vitro performance of IP-PA. (A) Cellular uptake of IR783-cysteamine and IR-PA after incubation with CT26 cells for 6 h. Data shown as mean S.D., n = 3. The values are compared via unpaired t-test. **p o 0.002. (B) Cellular uptake of IR-PA in CT26 cells after different treatments. Data shown as mean S.D., n = 3. The values are compared with the non-treated group via one-way ANOVA. **p o 0.002, *** p o 0.0002. (C) Representative fluorescence images of CT26 cells after incubating with IR-PA for 6 h (red: IR-PA, green: mitochondria stained by MitoTracker, yellow: pixels with co- localized red and green colours). Scale bar = 20 mm. Quantitative co-localization analysis of red and green signals is performed by correlation regression between red and green intensities shown in the scatterplot at the upper right corner. Qualitative co-localization analysis is visualized by the plot profile (lower right corner) along the direction of the white arrow in the microscopic image. (D) Representative fluorescence images of CT26 cells incubated with CM-H2DCFDA probes after different treatment (green: activated CM-H2DCFDA probes indicating generated ROS). Scale bar = 100 mm. (E) Mean green fluorescence intensity from the images in (D). Data shown as mean S.D., n = 3. The values are compared via unpaired t-test. **p o 0.002, *** p o 0.0002. (F) Live/dead (calcein/EthD-1) cytotoxic assay staining of CT26 cells after different treatments (green: calcein staining indicating living cells, red: EthD-1 staining indicating dead cells). Scale bar = 100 mm. (G) Quantified result showing the dead cell percentage from the experiment presented in Fig. 2D. Data shown as mean S.D., n = 4. The values are compared via unpaired t-test. *p o 0.03, *** p o 0.0002. (H) Cytotoxicity of different treatments measured by CCK-8 kit with CT26 cells. Data shown as mean S.D., n = 3. The values are compared via unpaired t-test, **p o 0.002, **** p o 0.0001.

Article Snippet: IR783 (sodium salt), triethylamine (TEA), ethyl acetate, and 1,3- diphenylisobenzofuran (DPBF) were purchased from Tokyo Chemical Industry Co., Ltd (Tokyo, Japan).

Techniques: In Vitro, Incubation, Fluorescence, Staining, Generated, CCK-8 Assay