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‘In Vitro’, ‘In Vivo’ and ‘In Silico’ Investigation of the Anticancer Effectiveness of Oxygen-Loaded Chitosan-Shelled Nanodroplets as Potential Drug Vector

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ABSTRACT

Purpose

Chitosan-shelled/decafluoropentane-cored oxygen-loaded nanodroplets (OLN) are a new class of nanodevices to effectively deliver anti-cancer drugs to tumoral cells. This study investigated their antitumoral effects ‘per se’, using a mathematical model validated on experimental data.

Methods

OLN were prepared and characterized either in vitro or in vivo. TUBO cells, established from a lobular carcinoma of a BALB-neuT mouse, were investigated following 48 h of incubation in the absence/presence of different concentrations of OLN. OLN internalization, cell viability, necrosis, apoptosis, cell cycle and reactive oxygen species (ROS) production were checked as described in the Method section.

In vivo tumor growth was evaluated after subcutaneous transplant in BALB/c mice of TUBO cells either without treatment or after 24 h incubation with 10% v/v OLN.

Results

OLN showed sizes of about 350 nm and a positive surface charge (45 mV). Dose-dependent TUBO cell death through ROS-triggered apoptosis following OLN internalization was detected. A mathematical model predicting the effects of OLN uptake was validated on both in vitro and in vivo results.

Conclusions

Due to their intrinsic toxicity OLN might be considered an adjuvant tool suitable to deliver their therapeutic cargo intracellularly and may be proposed as promising combined delivery system.

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Abbreviations

DAPI:

Diamidino-2-phenylindole dihydrochloride

DMEM:

Dulbecco’s modified eagle medium

FITC:

Fluorescein isothiocyanate

LDH:

Lactate dehydrogenase

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OLN:

Oxygen-loaded Nanodroplets

ROS:

Oxygen Reactive Species

References

  1. Talevi A, Gantner ME, Ruiz ME. Applications of nanosystems to anticancer drug therapy (Part I. Nanogels, nanospheres, nanocapsules). Recent Pat Anticancer Drug Discov. 2014;9(1):83–98.

    Article  CAS  PubMed  Google Scholar 

  2. Ferrari M. Nanovector therapeutics. Curr Opin Chem Biol. 2005;9(4):343–6.

    Article  CAS  PubMed  Google Scholar 

  3. Gao H, Shi W, Freund LB. Mechanics of receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 2005;102(27):9469–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Mora-Huertas CE, Fessi H, Elaissari A. Polymer-based nanocapsules for drug delivery. Int J Pharm. 2010;385(1–2):113–42.

    Article  CAS  PubMed  Google Scholar 

  5. Barrera G, Daga M, Ferrara B, Dianzani C, Pizzimenti S, Argenziano M, et al. Drug delivery nanoparticles in treating Chemoresistant tumor cells. Curr Med Chem. 2017;24(42):4800–15.

    CAS  PubMed  Google Scholar 

  6. Lin Q, Bao C, Yang Y, Liang Q, Zhang D, Cheng S, et al. Highly discriminating photorelease of anticancer drugs based on hypoxia activatable phototrigger conjugated chitosan nanoparticles. Adv Mater. 2013;25(14):1981–6.

    Article  CAS  PubMed  Google Scholar 

  7. Magnetto C, Prato M, Khadjavi A, Giribaldi G, Fenoglio I, Jose J, et al. Ultrasound-activated decafluoropentane-cored and chitosan-shelled nanodroplets for oxygen delivery to hypoxic cutaneous tissues. RSC Adv. 2014;4(72):38433–41.

    Article  CAS  Google Scholar 

  8. Prato M, Magnetto C, Jose J, Khadjavi A, Cavallo F, Quaglino E, et al. 2H,3H-decafluoropentane-based nanodroplets: new perspectives for oxygen delivery to hypoxic cutaneous tissues. PLoS One. 2015;10(3):e0119769.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Cavalli R, Bisazza A, Giustetto P, Civra A, Lembo D, Trotta G, et al. Preparation and characterization of dextran nanobubbles for oxygen delivery. Int J Pharm. 2009;381(2):160–5.

    Article  CAS  PubMed  Google Scholar 

  10. Cavalli R, Bisazza A, Rolfo A, Balbis S, Madonnaripa D, Caniggia I, et al. Ultrasound-mediated oxygen delivery from chitosan nanobubbles. Int J Pharm. 2009;378(1–2):215–7.

    Article  CAS  PubMed  Google Scholar 

  11. Banche G, Prato M, Magnetto C, Allizond V, Giribaldi G, Argenziano M, et al. Antimicrobial chitosan nanodroplets: new insights for ultrasound-mediated adjuvant treatment of skin infection. Future Microbiol. 2015;10(6):929–39.

    Article  CAS  PubMed  Google Scholar 

  12. Khadjavi A, Magnetto C, Panariti A, Argenziano M, Gulino GR, Rivolta I, et al. Chitosan-shelled oxygen-loaded nanodroplets abrogate hypoxia dysregulation of human keratinocyte gelatinases and inhibitors: new insights for chronic wound healing. Toxicol Appl Pharmacol. 2015;286(3):198–206.

    Article  CAS  PubMed  Google Scholar 

  13. Rovero S, Amici A, Carlo ED, Bei R, Nanni P, Quaglino E, et al. DNA vaccination against rat her-2/Neu p185 more effectively inhibits carcinogenesis than transplantable carcinomas in transgenic BALB/c mice. J Immunol. 2000;165(9):5133–42.

    Article  CAS  PubMed  Google Scholar 

  14. Minero VG, Khadjavi A, Costelli P, Baccino FM, Bonelli G. JNK activation is required for TNFalpha-induced apoptosis in human hepatocarcinoma cells. Int Immunopharmacol. 2013;17(1):92–8.

    Article  CAS  PubMed  Google Scholar 

  15. Salvati A, Åberg C, dos Santos T, Varela J, Pinto P, Lynch I, et al. Experimental and theoretical comparison of intracellular import of polymeric nanoparticles and small molecules: toward models of uptake kinetics. Nanomedicine. 2011;7(6):818–26.

    Article  CAS  PubMed  Google Scholar 

  16. Maher MA, Naha PC, Mukherjee SP, Byrne HJ. Numerical simulations of in vitro nanoparticle toxicity - the case of poly(amido amine) dendrimers. Toxicol in Vitro. 2014;28(8):1449–60.

    Article  CAS  PubMed  Google Scholar 

  17. Souto GD, Farhane Z, Casey A, Efeoglu E, McIntyre J, Byrne HJ. Evaluation of cytotoxicity profile and intracellular localisation of doxorubicin-loaded chitosan nanoparticles. Anal Bioanal Chem. 2016;408(20):5443–55.

    Article  CAS  PubMed  Google Scholar 

  18. Kanapathipillai M, Brock A, Ingber DE. Nanoparticle targeting of anti-cancer drugs that alter intracellular signaling or influence the tumor microenvironment. Adv Drug Deliv Rev. 2014;79-80:107–18.

    Article  CAS  PubMed  Google Scholar 

  19. Duchene D, Cavalli R, Gref R. Cyclodextrin-based polymeric nanoparticles as efficient carriers for anticancer drugs. Curr Pharm Biotechnol. 2016;17(3):248–55.

    Article  CAS  PubMed  Google Scholar 

  20. Li Q, Dunn ET, Grandmaison EW, Goosen MFA. Applications and properties of chitosan. J Bioact Compat Polym. 1992;7(4):370–97.

    Article  CAS  Google Scholar 

  21. Kumar MNVR, Muzzarelli RAA, Muzzarelli C, Sashiwa H, Domb AJ. Chitosan chemistry and pharmaceutical perspectives. Chem Rev. 2004;104(12):6017–84.

    Article  PubMed  Google Scholar 

  22. Qi LF, Xu ZR, Li Y, Jiang X, Han XY. In Vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells. World J Gastroenterol. 2005;11(33):5136–41.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Qi LF, Xu ZR. In vivo antitumor activity of chitosan nanoparticles. Bioorg Med Chem Lett. 2006;16(16):4243–5.

    Article  CAS  PubMed  Google Scholar 

  24. Jiang M, Ouyang H, Ruan P, Zhao H, Pi Z, Huang S, et al. Chitosan derivatives inhibit cell proliferation and induce apoptosis in breast cancer cells. Anticancer Res. 2011;31(4):1321–8.

    CAS  PubMed  Google Scholar 

  25. Salah R, Michaud P, Mati F, Harrat Z, Lounici H, Abdi N, et al. Anticancer activity of chemically prepared shrimp low molecular weight chitin evaluation with the human monocyte leukaemia cell line, THP-1. Int J Biol Macromol. 2013;52:333–9.

    Article  CAS  PubMed  Google Scholar 

  26. Xu YL, et al. Chitosan nanoparticles inhibit the growth of human hepatocellular carcinoma xenografts through an antiangiogenic mechanism. Anticancer Res. 2009;29(12):5103–9.

    CAS  PubMed  Google Scholar 

  27. Hasegawa M, Yagi K, Iwakawa S, Hirai M. Chitosan induces apoptosis via caspase-3 activation in bladder tumor cells. Jpn J Cancer Res. 2001;92(4):459–66.

    Article  CAS  PubMed  Google Scholar 

  28. Takimoto H, Hasegawa M, Yagi K, Nakamura T, Sakaeda T, Hirai M. Proapoptotic effect of a dietary supplement: water soluble chitosan activates caspase-8 and modulating death receptor expression. Drug Metab Pharmacokinet. 2004;19(1):76–82.

    Article  CAS  PubMed  Google Scholar 

  29. Amidi M, Mastrobattista E, Jiskoot W, Hennink WE. Chitosan-based delivery systems for protein therapeutics and antigens. Adv Drug Deliv Rev. 2010;62(1):59–82.

    Article  CAS  PubMed  Google Scholar 

  30. Kean T, Thanou M. Biodegradation, biodistribution and toxicity of chitosan. Adv Drug Deliv Rev. 2010;62(1):3–11.

    Article  CAS  PubMed  Google Scholar 

  31. Cavalli R, Leone F, Minelli R, Fantozzi R, Dianzani C. New chitosan nanospheres for the delivery of 5-fluorouracil: preparation, characterization and in vitro studies. Curr Drug Deliv. 2014;11(2):270–8.

    Article  CAS  PubMed  Google Scholar 

  32. Wilhelm C, Billotey C, Roger J, Pons JN, Bacri JC, Gazeau F. Intracellular uptake of anionic superparamagnetic nanoparticles as a function of their surface coating. Biomaterials. 2003;24(6):1001–11.

    Article  CAS  PubMed  Google Scholar 

  33. Lee JK, Lim HS, Kim JH. Cytotoxic activity of aminoderivatized cationic chitosan derivatives. Bioorg Med Chem Lett. 2002;12(20):2949–51.

    Article  CAS  PubMed  Google Scholar 

  34. Edinger AL, Thompson CB. Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol. 2004;16(6):663–9.

    Article  CAS  PubMed  Google Scholar 

  35. Görlach A, Dimova EY, Petry A, Martínez-Ruiz A, Hernansanz-Agustín P, Rolo AP, et al. Reactive oxygen species, nutrition, hypoxia and diseases: problems solved? Redox Biol. 2015;6:372–85.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Dunn JD, et al. Reactive oxygen species and mitochondria: a nexus of cellular homeostasis. Redox Biol. 2015;6:472–85.

    Article  Google Scholar 

  37. Hsin YH, Chen CF, Huang S, Shih TS, Lai PS, Chueh PJ. The apoptotic effect of nanosilver is mediated by a ROS- and JNK-dependent mechanism involving the mitochondrial pathway in NIH3T3 cells. Toxicol Lett. 2008;179(3):130–9.

    Article  CAS  PubMed  Google Scholar 

  38. AshaRani PV, Low Kah Mun G, Hande MP, Valiyaveettil S. Cytotoxicity and genotoxicity of silver nanoparticles in human cells. ACS Nano. 2009;3(2):279–90.

    Article  CAS  PubMed  Google Scholar 

  39. Hussain SM, Hess KL, Gearhart JM, Geiss KT, Schlager JJ. In vitro toxicity of nanoparticles in BRL 3A rat liver cells. Toxicol in Vitro. 2005;19(7):975–83.

    Article  CAS  PubMed  Google Scholar 

  40. Limbach LK, Wick P, Manser P, Grass RN, Bruinink A, Stark WJ. Exposure of engineered nanoparticles to human lung epithelial cells: influence of chemical composition and catalytic activity on oxidative stress. Environ Sci Technol. 2007;41(11):4158–63.

    Article  CAS  PubMed  Google Scholar 

  41. Park EJ, Choi J, Park YK, Park K. Oxidative stress induced by cerium oxide nanoparticles in cultured BEAS-2B cells. Toxicology. 2008;245(1–2):90–100.

    Article  CAS  PubMed  Google Scholar 

  42. Kehrer JP, Klotz L. Free radicals and related reactive species as mediators of tissue injury and disease: implications for health. Crit Rev Toxicol. 2015;45(9):765–98.

    Article  CAS  PubMed  Google Scholar 

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Author information

Author notes

  1. Amina Khadjavi and Ilaria Stura contributed equally to this work.

Authors and Affiliations

  1. Dipartimento di Neuroscienze, Università di Torino, Corso Raffaello 30, 10125, Torino, Italy

    Amina Khadjavi & Caterina Guiot

  2. Dipartimento di Scienze della Sanità Pubblica e Pediatriche, Università di Torino, Via Santena 5 bis, Torino, 10126, Italy

    Ilaria Stura & Mauro Prato

  3. Dipartimento di Biotecnologie Molecolari e Scienze per la Salute, Università di Torino, Torino, Italy

    Valerio Giacomo Minero, Elena Quaglino & Federica Cavallo

  4. Dipartimento di Medicina Sperimentale, Università Milano Bicocca, Monza, Italy

    Alice Panariti & Ilaria Rivolta

  5. Dipartimento di Oncologia, Università di Torino, Torino, Italy

    Giulia Rossana Gulino & Giuliana Giribaldi

  6. Dipartimento di Scienze e Tecnologia del Farmaco, Università di Torino, Torino, Italy

    Federica Bessone & Roberta Cavalli

Authors
  1. Amina Khadjavi
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  2. Ilaria Stura
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  3. Mauro Prato
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  4. Valerio Giacomo Minero
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  8. Federica Bessone
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  10. Elena Quaglino
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  11. Roberta Cavalli
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  12. Federica Cavallo
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Corresponding author

Correspondence to Ilaria Stura.

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Khadjavi, A., Stura, I., Prato, M. et al. ‘In Vitro’, ‘In Vivo’ and ‘In Silico’ Investigation of the Anticancer Effectiveness of Oxygen-Loaded Chitosan-Shelled Nanodroplets as Potential Drug Vector. Pharm Res 35, 75 (2018). https://doi.org/10.1007/s11095-018-2371-z

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  • DOI: https://doi.org/10.1007/s11095-018-2371-z

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