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SCIENCE CHINA Materials, Volume 60, Issue 9: 866-880(2017) https://doi.org/10.1007/s40843-017-9079-6

A novel bone marrow targeted gadofullerene agent protect against oxidative injury in chemotherapy

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  • ReceivedJun 11, 2017
  • AcceptedJul 17, 2017
  • PublishedAug 14, 2017

Abstract

Chemotherapy as an effective cancer treatment technique has been widely used in tumor therapy. However, it is still a challenge to overcome the serious side effects of chemotherapy, especially for its myelotoxicity. Here we report a novel strategy using the water soluble gadofullerene nanocrystals (GFNCs) to protect against chemotherapy injury in hepatocarcinoma bearing mice, which was induced by the commonly chemotherapeutic agent cyclophosphamide (CTX). The GFNCs were revealed to specifically accumulate in the bone marrow after intravenously injecting to mice and they exhibited excellent radical scavenging function, resulting in a prominent increase of mice blood cells and pathological improvements of the primary organs in the GFNCs (15 mg kg−1) treated mice after the CTX (60 mg kg−1) therapy. Moreover, the GFNCs maintained and even strengthened the antineoplastic activity of the CTX agent. Therefore, the GFNCs would be the promising chemoprotective agents in chemotherapy based on their high efficiency, low toxicity and metabolizable property.


Funded by

National Natural Science Foundation of China(51472248,51372251,51502301)

National Major Scientific Instruments and Equipments Development Project(ZDYZ2015-2)

Key Research Program of the Chinese Academy of Sciences(QYZDJ-SSW-SLH025,KGZD-EW-T02,XDA09030302)


Acknowledgment

This work was financially supported by the National Natural Science Foundation of China (51472248, 51372251 and 51502301), the National Major Scientific Instruments and Equipments Development Project (ZDYZ2015-2), and the Key Research Program of the Chinese Academy of Sciences (QYZDJ-SSW-SLH025, KGZD-EW-T02 and XDA09030302).


Interest statement

The authors declare that they have no conflict of interest.


Contributions statement

Zhang Y, Zhen M and Wang C performed the experiments, collected and analyzed the data, and wrote the manuscript; Shu C provided critical comments on the design of the study and the writing of the manuscript; Li J and Yu T performed the preparation of gadofullerenes nanocrystals (GFNCs); Jia W, Li X, Deng R, and Zhou Y provided essential technical assistance with experiments; Wang C conceived of and designed the study, supervised the research and wrote the manuscript. All authors discussed the results and approved the manuscript.


Author information

Ying Zhang was born in 1989. She received her PhD degree in physical chemistry from the Institute of Chemistry, Chinese Academy of Sciences (ICCAS) in 2017. Her research interests include biomedical applications of fullerenes and gadofullerenes.


Mingming Zhen was born in 1987. She received her PhD degree in physical chemistry from the ICCAS in 2014. Currently, she is an assistant professor at the ICCAS. Her research interests include biomedical applications of fullerenes and gadofullerenes.


Chunru Wang was born in 1965. He received his PhD degree in physical chemistry from Dalian Institute of Chemistry Physics, Chinese Academy of Sciences in 1992. Currently, he is a professor at the ICCAS. His research interests include fullerenes and endohedral fullerenes, mainly focusing on their industrialization and applications. He discovered the metal carbide fullerenes for the first time, researched on high efficiency MRI contrast agents and developed a novel tumor vascular-targeting therapy technique using gadofullerenes.


Supplement

Supplementary information

Experimental details and supporting data are available in the online version of the paper.


References

[1] André N, Carré M, Pasquier E. Metronomics: towards personalized chemotherapy?. Nat Rev Clin Oncol, 2014, 11: 413-431 CrossRef PubMed Google Scholar

[2] Kurtova AV, Xiao J, Mo Q, et al. Blocking PGE2-induced tumour repopulation abrogates bladder cancer chemoresistance. Nature, 2014, 517: 209-213 CrossRef PubMed ADS Google Scholar

[3] Fu D, Calvo JA, Samson LD. Balancing repair and tolerance of DNA damage caused by alkylating agents. Nat Rev Cancer, 2012, 52 CrossRef PubMed Google Scholar

[4] Helleday T, Petermann E, Lundin C, et al. DNA repair pathways as targets for cancer therapy. Nat Rev Cancer, 2008, 8: 193-204 CrossRef PubMed Google Scholar

[5] Costa L, Major PP. Effect of bisphosphonates on pain and quality of life in patients with bone metastases. Nat Clin Prac Oncol, 2009, 6: 163-174 CrossRef PubMed Google Scholar

[6] Lachmann N, Czarnecki K, Brennig S, et al. Deoxycytidine-kinase knockdown as a novel myeloprotective strategy in the context of fludarabine, cytarabine or cladribine therapy. Leukemia, 2015, 29: 2266-2269 CrossRef PubMed Google Scholar

[7] Das UB, Mallick M, Debnath JM, Ghosh D. Protective effect of ascorbic acid on cyclophosphamide-induced testicular gametogenic and androgenic disorders in male rats. Asian J androl, 2002, 4: 201–207. Google Scholar

[8] Lucas D, Scheiermann C, Chow A, et al. Chemotherapy-induced bone marrow nerve injury impairs hematopoietic regeneration. Nat Med, 2013, 19: 695-703 CrossRef PubMed Google Scholar

[9] Levesque JP, Winkler IG. It takes nerves to recover from chemotherapy. Nat Med, 2013, 19: 669-671 CrossRef PubMed Google Scholar

[10] Trachootham D, Alexandre J, Huang P. Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach?. Nat Rev Drug Discov, 2009, 8: 579-591 CrossRef PubMed Google Scholar

[11] Dvash E, Har-Tal M, Barak S, et al. Leukotriene C4 is the major trigger of stress-induced oxidative DNA damage. Nat Commun, 2015, 6: 10112 CrossRef PubMed ADS Google Scholar

[12] Macleod KF. The role of the RB tumour suppressor pathway in oxidative stress responses in the haematopoietic system. Nat Rev Cancer, 2008, 8: 769-781 CrossRef PubMed Google Scholar

[13] Lin W, Yuan N, Wang Z, et al. Autophagy confers DNA damage repair pathways to protect the hematopoietic system from nuclear radiation injury. Sci Rep, 2015, 5: 12362-12373 CrossRef PubMed ADS Google Scholar

[14] Kerbel RS, Kamen BA. The anti-angiogenic basis of metronomic chemotherapy. Nat Rev Cancer, 2004, 4: 423-436 CrossRef PubMed Google Scholar

[15] Manda K, Bhatia AL. Prophylactic action of melatonin against cyclophosphamide-induced oxidative stress in mice. Cell Biol Toxicol, 2003, 19: 367-372 CrossRef Google Scholar

[16] Patra K, Bose S, Sarkar S, et al. Amelioration of cyclophosphamide induced myelosuppression and oxidative stress by cinnamic acid. Chemico-Biol Interactions, 2012, 195: 231-239 CrossRef PubMed Google Scholar

[17] Xue Y, Lim S, Yang Y, et al. PDGF-BB modulates hematopoiesis and tumor angiogenesis by inducing erythropoietin production in stromal cells. Nat Med, 2011, 18: 100-110 CrossRef PubMed Google Scholar

[18] Passegué E, Ernst P. IFN-α wakes up sleeping hematopoietic stem cells. Nat Med, 2009, 15: 612-613 CrossRef PubMed Google Scholar

[19] Finkel T. Oxidant signals and oxidative stress. Curr Opin Cell Biol, 2003, 15: 247-254 CrossRef Google Scholar

[20] Winterbourn C. Oxidative denaturation in congenital hemolytic anemias: the unstable hemoglobins. Semin Hematol, 1990, 27: 41–50. Google Scholar

[21] Lee J, Lim KT. Protection against cyclophosphamide-induced myelosuppression by ZPDC glycoprotein (24 kDa). Immunol Invest, 2013, 42: 61-80 CrossRef PubMed Google Scholar

[22] Kawano M, Mabuchi S, Matsumoto Y, et al. The significance of G-CSF expression and myeloid-derived suppressor cells in the chemoresistance of uterine cervical cancer. Sci Rep, 2016, 5: 18217 CrossRef PubMed ADS Google Scholar

[23] Bakanay M, Demirer T. Novel agents and approaches for stem cell mobilization in normal donors and patients. Bone Marrow Transplant, 2012, 47: 1154-1163 CrossRef PubMed Google Scholar

[24] Afifi S, Adel NG, Devlin S, et al. Upfront plerixafor plus G-CSF versus cyclophosphamide plus G-CSF for stem cell mobilization in multiple myeloma: efficacy and cost analysis study. Bone Marrow Transplant, 2016, 51: 546-552 CrossRef PubMed Google Scholar

[25] Barreto JA, O'Malley W, Kubeil M, et al. Nanomaterials: applications in cancer imaging and therapy. Adv Mater, 2011, 23: H18-H40 CrossRef PubMed Google Scholar

[26] Ferrari M. Cancer nanotechnology: opportunities and challenges. Nat Rev Cancer, 2005, 5: 161-171 CrossRef PubMed Google Scholar

[27] Wicki A, Witzigmann D, Balasubramanian V, et al. Nanomedicine in cancer therapy: challenges, opportunities, and clinical applications. J Control Release, 2015, 200: 138-157 CrossRef PubMed Google Scholar

[28] Sayes CM, Marchione AA, Reed KL, et al. Comparative pulmonary toxicity assessments of C60 water suspensions in rats:  few differences in fullerene toxicity in vivo in contrast to in vitro profiles. Nano Lett, 2007, 7: 2399-2406 CrossRef PubMed ADS Google Scholar

[29] Aschberger K, Johnston HJ, Stone V, et al. Review of fullerene toxicity and exposure—appraisal of a human health risk assessment, based on open literature. Regul Toxicol Pharmacol, 2010, 58: 455-473 CrossRef PubMed Google Scholar

[30] Yan L, Zhao F, Li S, et al. Low-toxic and safe nanomaterials by surface-chemical design, carbon nanotubes, fullerenes, metallofullerenes, and graphenes. Nanoscale, 2011, 3: 362-382 CrossRef PubMed ADS Google Scholar

[31] Wang J, Chen C, Li B, et al. Antioxidative function and biodistribution of [Gd@C82(OH)22]n nanoparticles in tumor-bearing mice. Biochem Pharmacol, 2006, 71: 872-881 CrossRef PubMed Google Scholar

[32] Zhen M, Shu C, Li J, et al. A highly efficient and tumor vascular-targeting therapeutic technique with size-expansible gadofullerene nanocrystals. Sci China Mater, 2015, 58: 799-810 CrossRef Google Scholar

[33] Cagle DW, Kennel SJ, Mirzadeh S, et al. In vivo studies of fullerene-based materials using endohedral metallofullerene radiotracers. Proc Natl Acad Sci USA, 1999, 96: 5182-5187 CrossRef ADS Google Scholar

[34] Gharbi N, Pressac M, Hadchouel M, et al. Fullerene is a powerful antioxidant in vivo with no acute or subacute toxicity. Nano Lett, 2005, 5: 2578-2585 CrossRef PubMed ADS Google Scholar

[35] Yin JJ, Lao F, Fu PP, et al. The scavenging of reactive oxygen species and the potential for cell protection by functionalized fullerene materials. Biomaterials, 2009, 30: 611-621 CrossRef PubMed Google Scholar

[36] Duarte JH. Experimental arthritis: fullerene nanoparticles ameliorate disease in arthritis mouse model. Nat Rev Rheumatol, 2015, 11: 319-319 CrossRef PubMed Google Scholar

[37] Xu JY, Su YY, Cheng JS, et al. Protective effects of fullerenol on carbon tetrachloride-induced acute hepatotoxicity and nephrotoxicity in rats. Carbon, 2010, 48: 1388-1396 CrossRef Google Scholar

[38] Milic VD, Stankov K, Injac R, et al. Activity of antioxidative enzymes in erythrocytes after a single dose administration of doxorubicin in rats pretreated with fullerenol C60(OH)24. Toxicol Mech Methods, 2009, 19: 24-28 CrossRef PubMed Google Scholar

[39] Baati T, Bourasset F, Gharbi N, et al. The prolongation of the lifespan of rats by repeated oral administration of fullerene. Biomaterials, 2012, 33: 4936-4946 CrossRef PubMed Google Scholar

[40] Ji ZQ, Sun H, Wang H, et al. Biodistribution and tumor uptake of C60(OH)x in mice. J Nanopart Res, 2006, 8: 53-63 CrossRef ADS Google Scholar

[41] Zheng J, Zhen MM, Ge JC, et al. Multifunctional gadofulleride nanoprobe for magnetic resonance imaging/fluorescent dual modality molecular imaging and free radical scavenging. Carbon, 2013, 65: 175-180 CrossRef Google Scholar

[42] Markovic Z, Trajkovic V. Biomedical potential of the reactive oxygen species generation and quenching by fullerenes (C60). Biomaterials, 2008, 29: 3561-3573 CrossRef PubMed Google Scholar

[43] Andrievsky GV, Bruskov VI, Tykhomyrov AA, et al. Peculiarities of the antioxidant and radioprotective effects of hydrated C60 fullerene nanostuctures in vitro and in vivo. Free Radical Biol Med, 2009, 47: 786-793 CrossRef PubMed Google Scholar

[44] Karakoti A, Singh S, Dowding JM, et al. Redox-active radical scavenging nanomaterials. Chem Soc Rev, 2010, 39: 4422-4432 CrossRef PubMed Google Scholar

[45] Lee HJ, Selesniemi K, Niikura Y, et al. Bone marrow transplantation generates immature oocytes and rescues long-term fertility in a preclinical mouse model of chemotherapy-induced premature ovarian failure. J Clin Oncol, 2007, 25: 3198-3204 CrossRef PubMed Google Scholar

[46] Wang H, Agarwal P, Zhao S, et al. Combined cancer therapy with hyaluronan-decorated fullerene-silica multifunctional nanoparticles to target cancer stem-like cells. Biomaterials, 2016, 97: 62-73 CrossRef PubMed Google Scholar

[47] Sun M, Kiourti A, Wang H, et al. Enhanced microwave hyperthermia of cancer cells with fullerene. Mol Pharm, 2016, 13: 2184-2192 CrossRef PubMed Google Scholar

[48] Naveiras O, Nardi V, Wenzel PL, et al. Bone-marrow adipocytes as negative regulators of the haematopoietic microenvironment. Nature, 2009, 460: 259-263 CrossRef PubMed ADS Google Scholar

[49] Cao S, Durrani FA, Tóth K, et al. Se-methylselenocysteine offers selective protection against toxicity and potentiates the antitumour activity of anticancer drugs in preclinical animal models. Br J Cancer, 2014, 110: 1733-1743 CrossRef PubMed Google Scholar

  • Figure 1

    Characteristics of GFNCs in vitro. (a) Schematic diagram of the as-synthesized GFNCs by hydroxylation and a photograph of the GFNCs aqueous solution. (b) AFM study of GFNCs to show the core size of the samples. (c) Average hydrodynamic diameter distributions of GFNCs in saline. (d) The ESR spectra of the hydroxyl radicals captured by DMPO after treatment with 50 μmol L−1 of GFNCs (red line). The saline was used as a blank (black line). The hydroxyl radical was generated by the H2O2 exposure to UV light for 8 min. (e) The cell viability of mice bone marrow FDC-P1 cells incubated with the GFNCs (0.5‒30 μmol L−1) for 24 h. It shows no toxicity of GFNCs towards the FDC-P1 cells. (f) Cytoprotective effects of the GFNCs (0.5‒30 μmol L−1) against H2O2-induced damage on the FDC-P1 cells (n = 6; *, P< 0.05).

  • Figure 2

    The biodistribution of GFNCs in vivo. (a) The GFNCs distribution in blood plasma of the test mice at different time points (5 min, 15 min,30 min, 45 min, 1 h, 4 h, and 24 h) after a single i.v. injection of GFNCs (expressed as ng Gd3+ mg−1 blood). (b) The GFNCs distribution in sclerotin and bone marrow of the test mice at 1 h and 24 h after administration with GFNCs (n = 5; *, P< 0.05). (c) Schematic illustration of the electron microprobe used to analyze the distribution of GFNCs in the mice bones. (d) The electron microprobe study about the time dependence of Gd3+ in mice bone marrow after GFNCs injection (0.5, 1, 4 and 24 h).

  • Figure 3

    The myelosuppression protective tests. (a) Schematic illustration of bone marrow protective process of the GFNCs. (b‒e) Hematological parameters including WBC, HGB, LY and MO% in the control group, the CTX group and the GFNCs + CTX group during the chemoprotective tests. (n = 6; *, P< 0.05). (f) Activities of MDA, SOD, GST, CAT and GPx in mice plasma on the day 18.

  • Figure 4

    The antitumor activity of CTX and the GFNCs protection against CTX-induced toxicity in mice. (a) The growth inhibitory curves of tumors (left) and body weight changes (right) in the control group, CTX group and GFNCs + CTX group (n = 6; *, P< 0.05, compared with the control group). (b) Photos of tumors of 18 tested mice after chemotherapy in three different groups. (c) Typical biochemical parameters: ALT, AST, ALP, LDH, BUN and UA, which are relevant to liver (ALT, AST and ALP), heart (LDH) and kidney functions (BUN and UA) (n = 6; *, P< 0.05). (d) Selected H&E sections of major organs (heart, liver, spleen, lung and kidney) and tumor tissues from the tested mice in different groups.

  • Figure 5

    Oxidative stress-related enzymes activities and H&E, ESEM analysis of mice tissues on the fourth day of CTX chemotherapy. (a) Activities of MDA, SOD, CAT, GPx, and GST in control, CTX, and GFNCs + CTX groups of mice (n = 4; *, P< 0.05). (b) Light microscopy of H&E sections of mice femur bone, in which the bone (Bo) and bone marrow (BM) labeled different bone sections; (► in red) labeled the normal bone marrow cells and hematopoietic cells, (► in black) labeled the fatty infiltration of hematopoietic tissues. (c) Light microscopy of H&E sections of mice spleen, in which (→ in black) labeled the splenic red pulp and (→ in red) the splenic white pulp. (d) ESEM images of mice bone marrow, in which (→ in blue) labeled the adipocytes and (→ in red) labeled the necrotic cells by the CTX injury. (e) ESEM images of mice blood red cells. Many shriveled and abnormal red cells were observed in the CTX group of mice, and the blood red cells in the GFNCs + CTX group showed normal morphology as that in controls.

  • Figure 6

    Oxidative stress-related enzyme activities and H&E, ESEM analyses of mice tissues on the eighth day of the CTX therapy. (a) Activities of MDA, SOD, CAT, GPx and GST in control, CTX, and GFNCs + CTX groups. (n = 4; *, P< 0.05). (b) Light microscopy of H&E sections of mice femur bone on the eighth day, in which the bone (Bo) and bone marrow (BM) labeled different bone sections; (► in red) labeled the normal bone marrow cells and hematopoietic cells, (► in black) labeled the fatty infiltration of hematopoietic tissues. (c) Light microscopy of H&E sections of mice spleen, in which (→ in black) labeled the splenic red pulp and (→ in red) the splenic white pulp. (d) ESEM images of mice bone marrow, in which (→ in blue) labeled the adipocytes and (→ in red) labeled the necrotic cells by the CTX injury. (e) ESEM images of mice blood red cells, in which the shriveled and abnormal red cells were mainly observed in the CTX group of mice.

  • Figure 7

    The schematic representation of the GFNCs protective mechanism related to bone marrow enrichment and free radical scavenging.

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