Multifunctional Reduced Graphene Oxide Hydrogel as Drug Carrier for Localized and Synergic Photothermal/Photodynamics/Chemo Therapy

doi:10.1016/j.jmst.2016.06.014

Abstract

Abstract:

To combine localized drug release with multimodal therapy for malignant tumor, a composite hydrogel as an integrative drug delivery system was facilely prepared. The system contains spinach extract (SE), reduced graphene oxide (rGO) and gold nanocages (AuNCs). SE conduces to the formation of hydrogel, and also serves as a green material for improving the biocompatibility of hydrogel, and a natural photosensitizer for killing tumor cells under laser radiation (660?nm). AuNCs show obvious photothermy and can enhance the generation of cytotoxic singlet oxygen (¹O₂). The composite hydrogel shell on tumor cells exhibits several competitive advantages including enhanced antitumor effect by retaining the high concentration of drugs around cancer cell, excellent PDT/PTT compatibility as well as high loading and controllable release of fluorouracil (5-FU) for synergetic multimodal treatment. The survival rate of HeLa cells incubated with 5-FU loaded hydrogel under NIR radiation for 10?min sharply decreases to 1.2%, indicating remarkably improved antitumor effects. These results demonstrate that the hydrogel is an excellent delivery carrier for localizable, NIR-responsive and combined PTT/PDT/Chemo synergetic antitumor.

Key words: Multimodal synergic therapy, Au nanocages, Spinach extract, Natural photosensitizer, Localized antitumor

Chang Guanru,Li Shikuo,Huang Fangzhi,Zhang Xiuzhen,Shen Yuhua,Xie Anjian. Multifunctional Reduced Graphene Oxide Hydrogel as Drug Carrier for Localized and Synergic Photothermal/Photodynamics/Chemo Therapy[J]. J. Mater. Sci. Technol., 2016, 32(8): 753-762.

Figures/Tables 9

Fig. 1. Illustration of the formation process, loaded with 5-FU and the antitumor mechanism of the composite hydrogel.

Fig. 2. (a) FTIR spectra of GO, rGO/AuNCs/SE hydrogel; (b) Raman spectra of GO, rGO/AuNCs/SE hydrogel; (c) XRD patterns of GO, rGO/AuNCs/SE and rGO/SE hydrogel; (d) XPS survey; (e) Au4f XPS spectrum and (f) C1s core level spectra of rGO/SE/AuNPs hydrogel, respectively.

Fig. 3. (a) SEM images of rGO/AuNCs/SE hydrogel and (b) element distribution of C, O and Au in rGO/AuNCs/SE hydrogel; TEM images of (c) GO, (d) and (e) rGO/AuNCs/SE hydrogel; (f) the selected area electron diffraction pattern of AuNCs.

Fig. 4. (a) SEM images of AuNCs; UV-vis-NIR absorption spectra of (b) AuNCs and (c) GO and rGO/AuNCs/SE; (d) IR thermal images of samples under NIR irradiation for 10?min and (e) temperature variation of samples at different time intervals when exposed to the NIR laser.

Fig. 5. Photobleaching of DPBF (9.0?×?10-5?M) via 1O2 generation in the presence of (a) SE, (b) rGO/SE precursor, (c) rGO/AuNCs/SE precursor, respectively; (d) time-dependent decrease in the absorbance at 410?nm via the oxidation of DPBF with the precursor of GO, SE, rGO/SE and rGO/AuNCs/SE under irradiation by a 660?nm laser. Error bars were based on the standard deviation of triplicate samples.

Fig. 6. Cumulative release profiles of 5-FU from hydrogel at pH of 7.4 and pH of 5.0 in PBS buffer at 37?°C.

Fig. 7. (a) HeLa cell viability assay of SE, rGO/SE and rGO/SE/AuNPs precursor at different concentrations (with the volume ratio of precursor and medium ranging from 1 to 0.01?mL/mL) and (b) viability assay of HeLa cells and CHO cells incubated with5-FU loaded rGO/AuNCs/SE hydrogels at different incubation period with or without irradiation (660?nm laser at a power density of 200 mW/cm2 for 10?min).

Fig. 8. Fluorescence microscopy images of HeLa cells incubated with rGO/SE precursor (a) without irradiation and (b) with irradiation for 10?min; HeLa cells incubated with rGO/AuNCs/SE precursor (c) before and (d) after irradiation for 10?min; HeLa cells incubated with rGO/AuNCs/SE precursor loaded with 5-FU (e) before and (f) after irradiation for 10?min; each series can be classified to cell nucleus dyed in blue by Hoechst 33342, red by PI and the merged images of both, respectively.

Fig. 9. (a) Optical and (b) fluorescence microscopy images of HeLa cells incubated with hydrogel shells; (c) the merge image of (a) and (b); straight 1 and 2 indicate HeLa cell and hydrogel shell, respectively; fluorescence microscopy images of (d) FITC-dextran with HeLa cells, (e) FITC-dextran labeled rGO/SE/AuNPs precursor and (f) rGO/AuNCs/SE hydrogel with HeLa cells.

References

[1]	A. Umar, B.K. Dunn, P. Greenwald. Nat.Rev. Cancer, 12(2012), pp. 835-848
[2]	S.H. Kaufmann, W.C. Earnshaw.Exp. Cell Res, 256(2000), pp. 42-49
[3]	Z.G. Zhou, B. Kong, C. Yu, X.Y. Shi, M.W. Wang, W. Liu, Y.N. Sun, Y.J. Zhang, H. Yang, S.P. Yang.Sci. Rep, 4(2014), pp. 3653-3662
[4]	Y. Yong, L.J. Zhou, Z.J. Gu, L. Yan, G. Tian, X.P. Zheng, X.D. Liu, X. Zhang, J.X. Shi, W.S. Cong, W.Y. Yin, Y.L. Zhao.Nanoscale, 6(2014), pp. 10394-10403
[5]	F. Li, K. Na.Biomacromolecules, 12(2011), pp. 1724-1730
[6]	Z. Chen, Z. Li, J. Wang, E. Ju, L. Zhou, J. Ren, X. Qu. Adv.Funct. Mater, 24(2014), pp. 522-529
[7]	Z. Wang, Z. Chen, Z. Liu, P. Shi, K. Dong, E. Ju, J. Ren, X. Qu.Biomaterials, 35(2014), pp. 9678-9688
[8]	L. Gao, J. Fei, J. Zhao, H. Li, Y. Cui, J. Li.ACS Nano, 6(2012), pp. 8030-8040
[9]	D. Phillips. Photochem.Photobiol. Sci, 9(2010), pp. 1589-1596
[10]	H. Zhang, C. Peng, J.Z. Yang, M. Lv, R. Liu, D.N. He, C.H. Fan, Q. Huang.ACS Appl. Mater. Interfaces, 5(2013), pp. 1761-1767
[11]	C. Zhu, Q. Yang, F. Lv, L. Liu, S. Wang. Adv.Mater, 25(2013), pp. 1203-1208
[12]	G.H. Wu, A. MikhaiLovsky, H.A. Khant, C. Fu, W. Chiu, J.A. Zasadzinski. J. Am. Chem.Soc, 130(2008), pp. 8175-8177
[13]	J.H. Park, G.V. Maltzahn, L.L. Ong, A. Centrone, T.A. Hatton, E. Ruoslahti, S.N. Bhatia, M.J. Sailor.Adv. Mater, 22(2010), pp. 880-885
[14]	S.T. Wang, K.J. Chen, T.H. Wu, H. Wang, W.Y. Lin, M. Ohashi, P.Y. Chiou, H.R. Tseng.Angew. Chem. Int. Ed. Engl, 49(2010), pp. 3777-3781
[15]	P. Shi, E. Ju, J. Ren, X. Qu. Adv.Funct. Mater, 24(2014), pp. 826-834
[16]	X. Yang, X. Liu, Z. Liu, F. Pu, J. Ren, X. Qu. Adv.Mater, 24(2012), pp. 2890-2895
[17]	Y.C. Wang, K.C.L.Black, L. Luehmann, W.Y. Li, Y. Zhang, X. Cai, D.H. Wan, S.Y. Liu, M. Li, P. Kim, Z.Y. Liu, L.V. Wang, Y.J. Liu, Y.N. Xia. ACS Nano, 7(2013), pp. 2068-2077
[18]	Y.T. Chen, F. Guo, Y. Qiu, H.R. Hu, I. Kulaots, E. Walsh, R.H. Hurt.ACS Nano, 7(2013), pp. 3744-3753
[19]	F. Li, S.J. Park, D. Ling, W. Park, J.Y. Han, K. Na, K. Char. J.Mater. Chem. B, 1(2013), pp. 1678-1686
[20]	K. Yang, J. Wan, S. Zhang, B. Tian, Y. Zhang, Z. Liu.Biomaterials, 33(2012), pp. 2206-2214
[21]	J.J. Castillo, T. Rindzevicius, L.V. Novoa, W.E. Svendsen, N. Rozlosnik, A. Boisen, P. Escobar, F. Martínez, J. Castillo-León. J. Mater. Chem.B, 1(2013), pp. 1475-1481
[22]	C.S. Jin, G. Zheng.Lasers Surg. Med, 43(2011), pp. 734-748
[23]	S.G. Shi, X.L. Zhu, Z.X. Zhao, W.J. Fang, M. Chen, Y.Z. Huang, X.L. Chen. J. Mater. Chem.B, 1(2013), pp. 1133-1141
[24]	L. Zhou, H. Jiang, S. Wei, X. Ge, J. Zhou, J. Shen.Carbon, 50(2012), pp. 5594-5604
[25]	S.A.M.Bergquist, U.E. Gertsson, M.E. Olsson. J. Sci. Food Agric, 86(2006), pp. 346-355
[26]	W.T. Li, H.W. Tsao, Y.Y. Chen, S.W. Cheng, Y.C. Hsu.Photochem. Photobiol. Sci, 6(2007), pp. 1341-1348
[27]	I. Gomaa, S.E. Ali, T.A.El-Tayeb, M.H. Abdel-Kader. PhotodiagnosisPhotodyn. Ther, 9(2012), pp. 362-368
[28]	X.J. Fan, G.Z. Jiao, L. Gao, P.F. Jin, X. Li. J.Mater. Chem. B, 1(2013), pp. 2658-2664
[29]	J.J. Castillo, T. Rindzevicius, L.V. Novoa, W.E. Svendsen, N. Rozlosnik, A. Boisen, P. Escobar, F. Martínez, J. Castillo-Leon. J. Mater. Chem.B, 1(2013), pp. 1475-1481
[30]	C.F. Hu, Y.L. Liu, J.L. Qin, G.T. Nie, B.F. Lei, Y. Xiao, M.T. Zheng, J.H. Rong.ACS Appl. Mater. Interfaces, 5(2013), pp. 4760-4768
[31]	Y.L. Wang, B.Z. Zhang, L. Zhu, Y.J. Li, F.Z. Huang, S.K. Li, Y.H. Shen, A.J. Xie.ACS Appl. Mater. Interfaces, 6(2014), pp. 15000-15006
[32]	G.R. Chang, Y.L. Wang, B.Y. Gong, Y.X. Xiao, Y. Chen, S.H. Wang, S.K. Li, F.Z. Huang, Y.H. Shen, A.J. Xie.ACS Appl. Mater. Interfaces, 7(2015), pp. 11246-11256
[33]	W.S. Hummers, R.E. Offeman. J. Am. Chem.Soc, 80(1958), p. 1339
[34]	S.E. Skrabalak, L. Au, X. Li, Y. Xia. Nat.Protoc, 2(2007), pp. 2182-2190
[35]	S.G. Awuah, J. Polreis, V. Biradar, Y. You. Org.Lett, 13(2011), pp. 3884-3887
[36]	B. Kundu, S.C. Kundu.Biomaterials, 33(2012), pp. 7456-7467
[37]	K. Takayama, H. Hirose, G. Tanaka, S. Pujals, S. Katayama, I. Nakase, S. Futaki. Mol.Pharm, 9(2012), pp. 1222-1230
[38]	Q. Liu, L. Wei, J. Wang, F. Peng, D. Luo, R. Cui, Y. Niu, X. Qin, Y. Liu, H. Sun, J. Yang, Y. Li.Nanoscale, 4(2012), pp. 7084-7089
[39]	H.X. Wu, H.L. Shi, Y.P. Wang, X.Q. Jia, C.Z. Tang, J.M. Zhang, S.P. Yang.Carbon, 69(2014), pp. 379-389
[40]	V. Singh, D. Joung, L. Zhai, S.S. Das, I. Khondaker, S. Seal. Prog.Mater. Sci, 56(2011), pp. 1178-1271
[41]	K. Krishnamoorthy, M. Veerapandian, K. Yun, S.J. Kim.Carbon, 53(2013), pp. 38-49
[42]	Y. Zhang, N. Zhang, Z.R. Tang, Y.J. Xu.ACS Nano, 6(2012), pp. 9777-9789
[43]	M. Turner, V.B. Golovko, O.P.H.Vaughan, P. Abdulkin, A.B. Murcia, M.S. Tikhov, B.F.G. Johnson, R.M. Lambert. Nature, 454(2008), pp. 981-983
[44]	N. Bouropoulos, S. Weiner, L. Addadi. Chem.Eur. J., 7(2001), pp. 1881-1888
[45]	X.Y. Yang, X.Y. Zhang, Z.F. Liu, Y.F. Ma, Y. Huang, Y.S. Chen. J. Phys. Chem.C, 112(2008), pp. 17554-17558
[46]	H. Pandey, V. Parashar, R. Parashar, R. Prakash, P.W. Ramteke, A.C. Pandey.Nanoscale, 3(2011), pp. 4104-4108
[47]	A. Ahsan, S.M. Hiniker, M.A. Davis, T.S. Lawrence, M.K. Nyati.Cancer Res, 69(2009), pp. 5108-5114
[48]	J. Zhao, Y. Huang, Y. Song, X. Zhao, J. Jin, J. Wang, L. Huang. Toxicol.Lett, 185(2009), pp. 124-131
[49]	X. Yang, G. Niu, X. Cao, Y. Wen, R. Xiang, H. Duan, Y. Chen. J.Mater. Chem, 22(2012), pp. 6649-6654