The Role of Crocetin-Loaded PLGA Nanoparticles as a Pre-Treatment Agent on Indocyanine-Photodynamic Therapy of Breast Cancer Cell

Document Type : Original Paper


1 Medical Physics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

2 Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences ,Mashhad, Iran


Introduction: Photodynamic therapy (PDT) can be considered as a non-invasive method for cancer treatment. One of the most commonly of a water-soluble dye photosensitizer (PS) used in photothermal therapy (PTT) and PDT is Indocyanine Green (ICG). However, high cytotoxicity in high concentration and instability in aqueous media were limited its application. It was shown that using nanoparticles or plant extracts in combination with PS could improve PDT efficiency. In this study, anti-cancer properties of crocetin (Crt) loaded PLGA (Poly lactic-co-glycolic acid) nanoparticles (NPs) were utilized to increase the PDT efficacy with ICG on the MCF-7 cells.
Material and Methods: Crt was encapsulated into PLGA NPs and its particle size distribution and encapsulation efficiency were evaluated. IC10 of Crt, PLGA-Crt NPs and ICG was determined by MTT assay in MCF-7 cancer cells. At these concentrations, the cells were pre-treated with Crt or PLG-Crt, then treated with ICG and finally exposure to near infrared (NIR) laser with 2.5 W powers at different times. The cells viability was evaluated by the MTT assay.
Results: The findings showed no dark cytotoxicity due to ICG (12.9 μM), Crt or PLGA-Crt alone. But NIR laser irradiation in the presence of ICG after cells pre-treatment by the Crt or PLGA-Crt NPs leads to induce cell death to (61.6 ±7) % and (75.5 ±5) %, respectively (P<0.05).
Conclusion: The results demonstrated that PLGA-Crt NPs in combination with ICG could improve PDT outcomes more efficiently in comparison with Crt and ICG. Therefore, this method could be effective in breast cancer therapy with low cytotoxicity.


Main Subjects

  1. Longo JPF, Muehlmann LA, Miranda-Vilela AL, Portilho FA, de Souza LR, Silva JR, et al. Prevention of Distant Lung Metastasis after Photodynamic Therapy Application in a Breast Cancer Tumor Model. J Biomed Nanotechnol. 2016;12(4):689-99.
  2. Brown SB, Brown EA, Walker I. The present and future role of photodynamic therapy in cancer treatment. Lancet Oncol. 2004;5(8):497-508.
  3. Villacorta RB, Roque KFJ, Tapang GA, Jacinto SD. Plant extracts as natural photosensitizers in photodynamic therapy: in vitro activity against human mammary adenocarcinoma MCF-7 cells. Asian Pac J Trop Biomed. 2017;7(4):358-66.
  4. Li B, Chu X, Gao M, Li W. Apoptotic mechanism of MCF-7 breast cells in vivo and in vitro induced by photodynamic therapy with C-phycocyanin. Acta Biochim Biophys Sin. 2010;42(1):80-9.
  5. Li L, Huh KM. Polymeric nanocarrier systems for photodynamic therapy. Biomater Res. 2014;18(1):19.
  6. Xiong W, Wang X, Hu J, Liu Y, Liu Q, Wang P. Comparative study of two kinds of repeated photodynamic therapy strategies in breast cancer by using a sensitizer, sinoporphyrin sodium. J Photochem Photobiol B. 2016;160:299-305.
  7. Khdair A, Chen D, Patil Y, Ma L, Dou QP, Shekhar MP, et al. Nanoparticle-mediated combination chemotherapy and photodynamic therapy overcomes tumor drug resistance. J Control Release. 2010;141(2):137-44.
  8. Sazgarnia A, Montazerabadi AR, Bahreyni-Toosi MH, Ahmadi A, Aledavood A. In vitro survival of MCF-7 breast cancer cells following combined treatment with ionizing radiation and mitoxantrone-mediated photodynamic therapy. Photodiagnosis Photodyn Ther. 2013;10(1):72-8.
  9. ZielińskaB A. Expression of Proapoptotic BAX and TP53 Genes and Antiapoptotic BCL-2 Gene in MCF-7 and T-47D Tumour Cell Cultures of the Mammary Gland After a Photodynamic Therapy with Photolon. Adv Clin Exp Med. 2015:37.
  10. Fan W, Huang P, Chen X. Overcoming the Achilles' heel of photodynamic therapy. Chem Soc Rev. 2016,45, 6488-6519.
  11. Ahn J-C, Kang J-W, Shin J-I, Chung P-S. Combination treatment with photodynamic therapy and curcumin induces mitochondria-dependent apoptosis in AMC-HN3 cells. Int J Oncol. 2012;41(6):2184-90.
  12. Sivasubramanian M, Chuang YC, Lo LW. Evolution of nanoparticle-mediated photodynamic therapy: From superficial to deep-seated cancers. Molecules. 2019 Jan;24(3):520.
  13. Boix-Garriga E, Acedo P, Casadó A, Villanueva A, Stockert JC, Cañete M, et al. Poly (D, L-lactide-co-glycolide) nanoparticles as delivery agents for photodynamic therapy: enhancing singlet oxygen release and photototoxicity by surface PEG coating. Nanotechnology. 2015;26(36):365104.
  14. El-Daly SM, Gamal-Eldeen AM, Abo-Zeid MA, Borai IH, Wafay HA, Abdel-Ghaffar A-RB. Photodynamic therapeutic activity of indocyanine green entrapped in polymeric nanoparticles. Photodiagnosis Photodyn Ther. 2013;10(2):173-85.
  15. Akbari T, Pourhajibagher M, Chiniforush N, Shahabi S, Hosseini F, Bahador A. Improve ICG based photodynamic properties through conjugation of icg into nano-graphene oxide against enterococcus faecalis. Avicenna J Clin Microbiol Infect. 2018;5(1): e64624.
  16. Zheng X, Zhou F, Wu B, Chen WR, Xing D. Enhanced tumor treatment using biofunctional indocyanine green-containing nanostructure by intratumoral or intravenous injection. Mol Pharm. 2012;9(3):514-22.
  17. Montazerabadi AR, Sazgarnia A, Bahreyni-Toosi MH, Ahmadi A, Aledavood A. The effects of combined treatment with ionizing radiation and indocyanine green-mediated photodynamic therapy on breast cancer cells. J Photochem Photobiol B. 2012;109:42-9.
  18. Ghorbani F, Attaran-Kakhki N, Sazgarnia A. The synergistic effect of photodynamic therapy and photothermal therapy in the presence of gold-gold sulfide nanoshells conjugated Indocyanine green on HeLa cells. Photodiagnosis Photodyn Ther. 2017;17:48-55.
  19. Skřivanová K, Škorpíková J, Švihálek J, Mornstein V, Janisch R. Photochemical properties of a potential photosensitiser indocyanine green in vitro. J Photochem Photobiol B. 2006;85(2):150-4.
  20. Kim SH, Lee JM, Kim SC, Park CB, Lee PC. Proposed cytotoxic mechanisms of the saffron carotenoids crocin and crocetin on cancer cell lines. Biochem Cell Biol. 2014;92(2):105-11.
  21. G Gutheil W, Reed G, Ray A, Anant S, Dhar A. Crocetin: an agent derived from saffron for prevention and therapy for cancer. Curr Pharm Biotechnol. 2012;13(1):173-9.
  22. Zhong Y-j, Shi F, Zheng X-l, Wang Q, Yang L, Sun H, et al. Crocetin induces cytotoxicity and enhances vincristine-induced cancer cell death via p53-dependent and-independent mechanisms. Acta Pharmacol Sin. 2011;32(12):1529-36.
  23. He K, Si P, Wang H, Tahir U, Chen K, Xiao J, et al. Crocetin induces apoptosis of BGC-823 human gastric cancer cells. Mol Med Rep. 2014;9(2):521-6.
  24. Pradhan J, Mohanty C, Sahoo SK. Protective efficacy of crocetin and its nanoformulation against cyclosporine A-mediated toxicity in human embryonic kidney cells. Life Sci. 2019;216:39-48.
  25. Hafezi Ghahestani Z, Alebooye Langroodi F, Mokhtarzadeh A, Ramezani M, Hashemi M. Evaluation of anti-cancer activity of PLGA nanoparticles containing crocetin. Artif Cells Nanomed Biotechnol. 2016:1-6.
  26. Langroodi F, Hafezi Ghahestani Z, Alibolandi M, Ebrahimian M, Hashemi M. Evaluation of the effect of crocetin on antitumor activity of doxorubicin encapsulated in PLGA nanoparticles. Nanomed J. 2016;3(1):23-34.
  27. Danhier F, Ansorena E, Silva JM, Coco R, Le Breton A, Préat V. PLGA-based nanoparticles: an overview of biomedical applications. J Control Release. 2012;161(2):505-22.
  28. Swed A, Cordonnier T, Fleury F, Boury F. Protein Encapsulation into PLGA Nanoparticles by a Novel Phase Separation Method Using Non-Toxic Solvents. J Nanomed Nanotechnol. 2014;5(241):2.
  29. Paszko E, Ehrhardt C, Senge MO, Kelleher DP, Reynolds JV. Nanodrug applications in photodynamic therapy. Photodiagnosis Photodyn Ther. 2011;8(1):14-29.
  30. Plackal Adimuriyil George B, Abrahamse H. A review on novel breast cancer therapies: Photodynamic therapy and plant derived agent induced cell death mechanisms. Anticancer Agents Med Chem. 2016;16(7):793-801.
  31. Marrelli M, Menichini G, Provenzano E, Conforti F. Applications of natural compounds in the photodynamic therapy of skin cancer. Curr Med Chem. 2014;21(12):1371-90.
  32. Gift MM, Ann KC, Ivan M-T, Heidi A. A review of nanoparticle photosensitizer drug delivery uptake systems for photodynamic treatment of lung cancer. Photodiagnosis Photodyn Ther. 2018;1(22):147-54.
  33. Saxena V, Sadoqi M, Shao J. Polymeric nanoparticulate delivery system for Indocyanine green: biodistribution in healthy mice. Int J Pharm. 2006;308(1):200-4.