Supporting information. Thermosensitive Lipid Bilayer-Coated Mesoporous Carbon. Nanoparticles for Synergistic Thermochemotherapy of Tumor

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1 Supporting information Thermosensitive Lipid Bilayer-Coated Mesoporous Carbon Nanoparticles for Synergistic Thermochemotherapy of Tumor Xian Li, Xiudan Wang, Luping Sha, Da Wang, Wei Shi, Qinfu Zhao*,, and Siling Wang*, School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang , China Shenyang No. 2 High School, 6 Wuai Street, Shenyang , China Corresponding authors Qinfu Zhao Phone: zqf @163.com Siling Wang Phone: silingwang@syphu.edu.cn S-1

2 Prescription optimization experiment In order to design the best prescription of liposome coverage, prescription screening experiment was conducted and details are stated as follows. MCN-COOH and liposomes were added at a weight ratio of 2:1, 1:1 and 1:2 to optimize the experiment. There are not any obvious differences from Zeta potential results (Figure S10). Nevertheless, the TSMCN with the ratio of both 1:1 and 1:2 had a better stability compared with the prescription of 2:1 (Figure S11). Therefore, the prescription of 1:1 was preferred from the economic angle. The weight ratio of liposome in TSMCN was 43.5 ± 1.65%, which was calculated by weight loss method. In detail, the TSMCN precipitate and the supernatant were collected and underwent freeze-drying process. The mass of liposome loaded in TSMCN was calculated by subtracting the amount in supernatant from the total added liposome. And the weight ratio of liposome in TSMCN was calculated according to the following equation: The weight ratio (%) = (total mass of liposome added the mass of liposome in supernatant) / (total mass of nanocarriers added + the mass of liposome loaded in the nanocarriers) 100. Hemolysis assays Rabbit blood stabilized with EDTA was provided by Animal Center of the Shenyang Pharmaceutical University. The whole blood was centrifuged to discard plasma and buffy coat and the red blood cells were then washed with PBS for several times. The resulting cells were diluted 10 times with sterile PBS solution. The 2 ml of the saline solutions of TSLP, MCN-COOH and TSMCN were added into the equal S-2

3 volume of rabbit red blood cells (RBCs) solutions, and the final concentrations of the samples were 20, 50, 100, 200, 500 and 1000 μg/ml. The mixtures let to rest for 4 h at room temperature after gently vortexing. Finally, the mixtures were centrifuged and the resulting solution of the upper layer was detected at 541 nm in a UV vis spectrophotometer. The hemolysis percentage of MCN-COOH and TSMCN was calculated by the following equation: Hemolytic percentage (%) = (Asample - Anegative control) / (Apositive control - Anegative control) 100 (A = the absorbance intensity of UV-Vis spectra) Table S1. The N 2 adsorption-desorption parameters of MCN, MCN-COOH and TSMCN. Sample S BET (m 2 /g) V t (cm 3 /g) D p (nm) MCN MCN-COOH TSMCN Figure S1. Raman spectrum of MCN-COOH. S-3

4 Figure S2. IR spectra of MCN, MCN-COOH and TSMCN. Figure S3. Dispersion stability of MCN-COOH and TSMCN at different times. S-4

5 Figure S4. (A) Hemolytic photographs and (B) hemolysis percentage of MCN-COOH and TSMCN at different concentrations (μg/ml). Figure S5. UV-vis-NIR absorption spectra of free DOX, liposome, TSMCN and DOX/TSMCN. Figure S6. IR thermal image of water and TSMCN suspensions under continuous NIR laser irradiation (1.25 W cm -2, 3 min). S-5

6 Figure S7. DSC curves of MCN-COOH, liposome and TSMCN. Figure S8. XPS spectra of MCN-COOH and TSMCN. S-6

7 Figure S9. Fluorescence images of 4T1 cells (A) irradiated by NIR laser and (B) incubated with 50 µg/ml TSMCN (scale bars = 50 μm). (C) and (D) represent the quantifiable analysis of the fluorescence in (A) and (B) using the ImageJ software, respectively. Figure S10. Heating curves of tumors of 4T1 mice treated with saline and TSMCN. S-7

8 Figure S11. Zeta potentials of TSMCN prepared at weight ratios of 2:1, 1:1 and 1:2 (MCN-COOH: liposome). Figure S12. Dispersion stabilities of TSMCN prepared at weight ratios of 2:1, 1:1 and 1:2 (MCN-COOH: liposome). S-8