Supporting Information

Transcription

1 Supporting Information Chemical Modifications of Porous Carbon Nanospheres Obtained from Ubiquitous Precursors for Targeted Drug Delivery and Live Cell Imaging Sutanu Kapri, Rahul Majee, and Sayan Bhattacharyya* Department of Chemical Sciences, and Centre for Advanced Functional Materials, Indian Institute of Science Education and Research (IISER) Kolkata, Mohanpur , India * for Correspondence: sayanb@iiserkol.ac.in Total number of pages: 16 Total number of Figures: 13 Total number of Tables: 3 Table of contents Item Description Page Table S1 List of common fluorescent dyes with their prices S2 Figure S1 FESEM images of PC-G, PC-H, PC-S, and PC-S-CTAB S3 Table S2 Results of elemental analysis of the porous carbon nanocarriers S3 Figure S2 Raman spectra of PCs S4 Figure S3 FTIR spectra of PCs S5 Figure S4 N 2 sorption isotherm and corresponding pore diameter distribution S6 (insets) of the PCs and PN-G-DOX Figure S5 Loading of PI onto PNs S7 Figure S6 Digital photograph of PC-G dispersion S8 Figure S7 Absorbance spectra of PN-G-DOX, PN-G-DOX/PEI and PN-G- S9 DOX/PEI-FA Figure S8 DLS spectra of the PNs before and after PI dye loading S10 Table S3 Hydrodynamic diameter and zeta potential of PNs before and after S11 PI loading Figure S9 Time dependent DOX release from PN-G-DOX and PN-G- S12 DOX/PEI Figure S10 Cell viability of different concentrations of PNs and PNs-PI against S13 HeLa cells, PN-G in MDA-MB-231 cells, respectively Figure S11 Cell viability of DOX, PN-G-DOX, PN-G-DOX/PEI and PN-G- S14 DOX/PEI-FA against RAW and J774A.1 cells Figure S12 Flow cytometry of PBS control, free DOX, PN-G-DOX/PEI and PN-G-DOX/PEI-DOX in HEK 293 cells S15 Figure S13 Quantification of fluorescence intensities of PI in HeLa cells S16 S1

2 Table S1: A list of common fluorescent dyes used for live cell imaging with their excitation (λ ex ) and emission (λ em ) wavelengths and base price from Sigma Aldrich. SL. No. Fluorescent Dyes λ ex (nm) λ em (nm) Base Price (US$)/mg Dyes with λ ex > 500 nm and λ em > 600 nm 1 Propidium Iodide (PI) Cy 5-azide Ethidiumhomodimer Cy 5 Maleimide Mono-Reactive Dye Dyes with λ ex > 500 nm and λ em < 600 nm 5 Rhodamine Nile Red > Tetramethylrhodamine isothiocyanate (TRITC) 8 Cy 3 Mono Dye Fluo Dyes with λ ex < 500 nm and λ em < 550 nm 10 Fluorescein 5-isothiocyanate FITC Hoechst ',6-diamidino-2-phenylindole (DAPI) CF 488A hydrazide Fura-2 AM Calcein-AM S2

3 Figure S1: FESEM images of (a) PC-G, (b) PC-H, (c) PC-S, and (d) PC-S-CTAB. Table S2: Results of elemental analysis of the porous carbon nanocarriers. Sample Carbon (wt%) Nitrogen (wt%) Hydrogen (wt%) Oxygen (calculated) (wt%) PN-G PN-H PN-S PN-S-CTAB PN-G-DOX PN-G-DOX/PEI PN-G-DOX/PEI-FA S3

4 Figure S2: Raman spectra of PCs obtained from different precursors. S4

5 Figure S3: FTIR spectra of PCs obtained from different precursors. S5

6 Figure S4: N 2 sorption isotherms and corresponding pore diameter distributions (inset) of (a) PC-G and PC-H, (b) PC-S and PC-S-CTAB and (c) PN-G-DOX, respectively. S6

7 Figure S5: Loading of PI onto PNs in PBS solution at ph 7.4. S7

8 Figure S6: Digital photographs of PC-G dispersion (water) and PN-G dispersion in water, PBS and DMEM media, respectively. S8

9 Figure S7: Absorbance spectra of PN-G-DOX, PN-G-DOX/PEI and PN-G-DOX/PEI-FA in PBS solution at ph 7.4. S9

10 Figure S8: DLS spectra of the PNs before and after PI dye loading (top and middle panel), and those of PN-G before and after DOX loading, PEI coating and FA functionalization (bottom panel). S10

11 Table S3: Hydrodynamic diameter and zeta potential of PNs before and after PI loading. Sample Hydrodynamic diameter (nm) Zeta potential (mv) PN-G PN-G-PI PN-H PN-H-PI PN-S PN-S-PI PN-S-CTAB PN-S-CTAB-PI S11

12 Figure S9: Time dependent DOX release from PN-G-DOX and PN-G-DOX/PEI in PBS buffer solutions at different ph and 37 o C for one week incubation. Data is represented as mean values ± standard deviation (SD) (n = 3), monitored by UV-vis spectroscopy. S12

13 Figure S10: (a) Cell viability of different concentrations of PNs against HeLa cells incubated for 48 h. (b) Cell viability of unbound PI, and different concentrations of PN-G-PI, PN-H-PI, PN-S-PI and PN-S-CTABA-PI against HeLa cells incubated for 48 h. (c) Cell viability of different concentrations of PN-G against MDA-MB-231 cells incubated for 48 h. S13

14 Figure S11: Cell viability of DOX, PN-G-DOX, PN-G-DOX/PEI and PN-G-DOX/PEI-FA against (a) RAW cells and (b) J774A.1 cells, incubated with different DOX concentrations for 24 h. Data is represented as mean values ± standard deviations (SD) (n = 3). S14

15 Figure S12: Quantitative analyses of DOX uptake by flow cytometry in HEK 293 cells where fluorescent intensity is shown in the x-axis and y-axis shows the cell counts. Cells were treated for 6 h with PBS control, free DOX, PN-G-DOX/PEI and PN-G-DOX/PEI-DOX. S15

16 Figure S13: Quantification of fluorescence intensities generated using ZEN lite software where average fluorescence intensities were calculated from 15 cells per sample. The intensities reflect the accumulation of PI inside the HeLa cells incubated with PI, PN-G-PI, PN-H-PI, PN-S-PI and PN-S-CTAB-PI for 12 h. S16