In vitro and in vivo evaluation of photo-induced antileishmanial activity of indocyanine green-loaded nanomicelles

Document Type : Original Article

Authors

1 Institute of Police Equipment and Technologies, Policing Sciences and Social Studies Research Institute, Tehran, Iran

2 Research Center of Advanced Technologies in Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran

3 Health Sciences Research Center, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran

4 Research Center for Life & Health Sciences & Biotechnology of the Police, Directorate of Health, Rescue & Treatment, Police Headquarters, Tehran, Iran

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

6 Department of Laboratory Sciences, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran

7 Department of Radiology Technology, School of Paramedical Sciences, Torbat Heydarieh University of Medical Sciences, Torbat Heydarieh, Iran

Abstract

Objective(s): Due to its low toxicity and high absorbance in the range of 650 to 900 nm, indocyanine green (ICG) has garnered significant attention for its applications in photodynamic therapy (PDT) and photothermal therapy (PTT). However, its tendency to aggregate in aqueous environments limits its efficacy in both in vitro and in vivo applications. Encapsulating ICG in a biocompatible nanomicelle can improve its aqueous stability and photophysical properties. The present study investigated the synergistic effect of ICG-loaded nanomicelles upon irradiation by an 808-nm laser on Leishmania major (L. major) parasites. 
Materials and Methods: Initially, a nanomicelle comprised ICG was synthesized and characterized. Then, the temperature increase during irradiation and promastigote viability were evaluated in vitro. Subsequently, the prepared samples’ in vitro dark toxicity and phototoxicity were assessed via the MTS assay. Finally, the in vivo antileishmanial efficacy of the ICG-loaded nanomicelles formulation was investigated in BALB/c mice. 
Results: The absorbance of ICG-loaded nanomicelles at 808 nm was more than 2 times greater than Free-ICG. Also, the prepared formulation exhibited a mean diameter of ~25 nm and a zeta potential of -2.3 ± 1 mV. The combination of ICG-loaded nanomicelles and 808 nm laser with a power density of 2.5 W cm−2 led to a significant reduction in the survival rate of promastigotes and lesion size of infected mice compared to control groups. 
Conclusion: The PDT/PTT mediated by ICG-loaded nanomicelles can be considered a promising and efficient therapeutic method for L. major, as it is inexpensive, safe, and easy to implement.

Keywords

Main Subjects


1. Akhzari S, Nabian S, Shayan P, Mazaheri Nezhad Fard R, Soltani M, Taheri M. Designing of RNA molecule translating for activitable melittin as selective targeting of Leishmania infected cells. Iran J Parasitol 2021; 16: 443-453.
2. Bilgic-Temel A, Murrell DF, Uzun S. Cutaneous leishmaniasis: A neglected disfiguring disease for women. Int J Womens Dermatol 2019; 5: 158-165.
3. Fernando A-S, Ana Lucia A-S, Kátia da Silva C, Flávia de Oliveira C. Introductory Chapter: Leishmania Parasites -Epidemiology and Immunopathogenesis. In: Fernando A-S, Flávia de Oliveira C, Ana Lucia A-S, Kátia da Silva C, editors. Leishmania Parasites. Rijeka: IntechOpen; 2024. p. Ch. 1
4. Firooz A, Mortazavi H, Khamesipour A, Ghiasi M, Abedini R, Balighi K, et al. Old world cutaneous leishmaniasis in Iran: Clinical variants and treatments. J Dermatolog Treat 2021; 32: 673-683.
5. Mohaghegh MA, Fata A, Salehi GH, Berenji F, Mousavi Bazzaz M, Rafatpanah H, et al. Molecular identification of Leishmania species using samples obtained from negative stained smears. Iran J Parasitol 2013; 8: 337-341.
6. Salehi G, Fata A, Mohaghegh MA, Mousavi Bazzaz SM, Rafatpanah H, Movahedi A. Molecular identification of Leishmania species in Taybad district, Iran. Asian Pac J Trop Dis 2014; 4: S535-S539.
7. Keyhani A, Sharifi I, Salarkia E, Khosravi A, Tavakoli Oliaee R, Babaei Z, et al. In vitro and in vivo therapeutic potentials of 6-gingerol in combination with amphotericin B for treatment of Leishmania major infection: Powerful synergistic and multifunctional effects. Int Immunopharmacol 2021; 101: 108274.
8. Kavian Z, Alavizadeh SH, Golmohamadzadeh S, Badiee A, Khamesipour A, Jaafari MR. Development of topical liposomes containing miltefosine for the treatment of Leishmania major infection in susceptible BALB/c mice. Acta Trop 2019; 196: 142-149.
9. Jaafari MR, Hatamipour M, Alavizadeh SH, Abbasi A, Saberi Z, Rafati S. Development of a topical liposomal formulation of Amphotericin B for the treatment of cutaneous leishmaniasis. Int J Parasitol Drugs Drug Resist 2019; 11: 156-165.
10. Aghaei  M, Aghaei  S, Sokhanvari  F, Ansari  N, Hosseini  SM, Mohaghegh MA, et al. The therapeutic effect of ozonated olive oil plus glucantime on human cutaneous leishmaniasis. Iran J Basic Med Sci 2019; 22: 25-30.
11. Sazgarnia A, Bahreyni-Toosi MH, Layegh P, Rajabi O, Ghodsinia RM. Liposomal zinc phthalocyanine as a potential agent for photodynamic therapy of leishmaniasis. Indian J Dermatol Venereol Leprol 2010; 76: 417-418.
12. Mayelifar K, Taheri AR, Rajabi O, Sazgarnia A. Ultraviolet B efficacy in improving antileishmanial effects of silver nanoparticles. Iran J Basic Med Sci 2015; 18: 677-683.
13. Shaddel M, Sharifi I, Karvar M, Keyhani A, Baziar Z. Cryotherapy of cutaneous leishmaniasis caused by Leishmania major in BALB/c mice: A comparative experimental study. J Vector Borne Dis 2018; 55: 42-46.
14. Berry S, Walker K, Hoskins C, Telling ND, Price HP. Nanoparticle-mediated magnetic hyperthermia is an effective method for killing the human-infective protozoan parasite Leishmania mexicana in vitro. Sci Rep 2019; 9: 1059-1067.
15. 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.
16. de Oliveira de Siqueira LB, da Silva Cardoso V, Rodrigues IA, Vazquez-Villa AL, Dos Santos EP, da Costa Leal Ribeiro Guimarães B, et al. Development and evaluation of zinc phthalocyanine nanoemulsions for use in photodynamic therapy for Leishmania spp. Nanotechnology 2017; 28: 065101.
17. Aureliano DP, Lindoso JAL, de Castro Soares SR, Takakura CFH, Pereira TM, Ribeiro MS. Cell death mechanisms in Leishmania amazonensis triggered by methylene blue-mediated antiparasitic photodynamic therapy. Photodiagnosis Photodyn Ther 2018; 23: 1-8.
18. Escobar, P, Vera AM, Neira LF, Velásquez AO, Carreño H. Photodynamic therapy using ultradeformable liposomes loaded with chlorine aluminum phthalocyanine against L. (V.) braziliensis experimental models. Exp Parasitol 2018; 194: 45-52.
19. Sepúlveda AAL, Arenas Velásquez AM, Patiño Linares IA, de Almeida L, Fontana CR, Garcia C, et al. Efficacy of photodynamic therapy using TiO(2) nanoparticles doped with Zn and hypericin in the treatment of cutaneous Leishmaniasis caused by Leishmania amazonensis. Photodiagnosis Photodyn Ther 2020; 30: 101676.
20. Marcolino LMC, Pereira AHC, Pinto JG, Mamone LA, Strixino JF. Cellular and metabolic changes after photodynamic therapy in leishmania promastigotes. Photodiagnosis Photodyn Ther 2021; 35: 102403.
21. Pereira AHC, Marcolino LMC, Pinto JG, Ferreira-Strixino J. Evaluation of the photodynamic therapy with curcumin on Leishmania braziliensis and Leishmania major amastigotes. Antibiotics (Basel) 2021; 10: 634-647.
22. El-Daly SM, Gamal-Eldeen AM, Abo-Zeid MA, Borai IH, Wafay HA, Abdel-Ghaffar AR. Photodynamic therapeutic activity of indocyanine green entrapped in polymeric nanoparticles. Photodiagnosis Photodyn Ther 2013; 10: 173-185.
23. Houthoofd S, Vuylsteke M, Mordon S, Fourneau I. Photodynamic therapy for atherosclerosis. The potential of indocyanine green. Photodiagnosis Photodyn Ther 2020; 29: 101568.
24. Topaloğlu N, Kadıköylü G, Onak G, Karaman O. The effect of indocyanine green-based photodynamic therapy on healthy fibroblast and keratinocyte cells. Photodiagnosis Photodyn Ther 2020; 31: 101891.
25. PaszkoE, Ehrhardt C, Senge MO, Kelleher DP, Reynolds JV. Nanodrug applications in photodynamic therapy. Photodiagnosis Photodyn Ther 2011; 8: 14-29.
26. Hamblin MR, Huang Y. Handbook of Photomedicine. 1st ed. 2013; 886.
27. Rai VK, Mishra N, Yadav KS, Yadav NP. Nanoemulsion as pharmaceutical carrier for dermal and transdermal drug delivery: Formulation development, stability issues, basic considerations and applications. J Control Release 2018; 270: 203-225.
28. Lucky SS, Soo KC, Zhang Y. Nanoparticles in photodynamic therapy. Chem Rev 2015; 115: 1990-2042.
29. Caldeira LR, Fernandes FR, Costa DF, Frézard F, Afonso LC,  Ferreira LA. Nanoemulsions loaded with amphotericin B: A new approach for the treatment of leishmaniasis. Eur J Pharm Sci 2015; 70: 125-131.
30. Sazgarnia A, Zabolinejad N, Layegh P, Rajabi O, Berenji F, Javidi Z, et al. Antileishmanial activity of liposomal clarithromycin against Leishmania major promastigotes. Iran J Basic Med Sci 2012; 15: 1210-1214.
31. Esfandiari F, Derakhshanfar A, Goudarzi F, Hatam G. Comparison of camel, dog and the laboratory animals’ sera with the fetal calf serum (FCS) for cultivation of Leishmania major. J Parasit Dis 2020; 44: 299-304.
32. Dolat E, Salarabadi SS, Layegh P, Jaafari MR, Sazgarnia S, Sazgarnia A. The effect of UV radiation in the presence of TiO(2)-NPs on Leishmania major promastigotes. Biochim Biophys Acta Gen Subj 2020; 1864: 129558.
33. Daneshvar F, Salehi F, Karimi M, Vais RD, Mosleh-Shirazi MA, Sattarahmady N. Combined X-ray radiotherapy and laser photothermal therapy of melanoma cancer cells using dual-sensitization of platinum nanoparticles. J Photochem Photobiol B 2020; 203: 111737.
34. Sazgarnia A, Taheri AR, Soudmand S, Parizi AJ, Rajabi O, Darbandi MS. Antiparasitic effects of gold nanoparticles with microwave radiation on promastigots and amastigotes of Leishmania major. Int J Hyperthermia 2013; 29: 79-86.
35. Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, Saffari M, Raoofian R, Hajjaran H, et al. Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol 2013; 135: 344-349.
36. Najm M, Badirzadeh A, Razmjou E, Alipour M, Khoshmirsafa M, Bahador A, et al. Photodynamic therapy using toluidine blue O (TBO) dye as a photosensitizer against Leishmania major. Iran J Public Health 2021; 50: 2111-2120.
37. Chen H, Wu L, Wang T, Zhang F, Song J, Fu J, et al. PTT/ PDT-induced microbial apoptosis and wound healing depend on immune activation and macrophage phenotype transformation. Acta Biomater 2023; 167: 489-505.
38. Kim H, Lee YR, Jeong H, Lee J, Wu X, Li H, et al. Photodynamic and photothermal therapies for bacterial infection treatment. Smart Molecules 2023; 1: e20220010.
39. Rodriguez VB, Henry SM, Hoffman AS, Stayton PS, Li X, Pun SH. Encapsulation and stabilization of indocyanine green within poly(styrene-alt-maleic anhydride) block-poly(styrene) micelles for near-infrared imaging. J Biomed Opt 2008; 13: 014025.
40. Kirchherr AK, Briel A, Mäder K. Stabilization of indocyanine green by encapsulation within micellar systems. Mol Pharm 2009; 6: 480-491.
41. Yan L and Qiu L. Indocyanine green targeted micelles with improved stability for near-infrared image-guided photothermal tumor therapy. Nanomedicine (Lond) 2015; 10: 361-373.
42. Roque L, Fernández M, Benito JM, Escudero I. Stability and characterization studies of Span 80 niosomes modified with CTAB in the presence of NaCl. Colloids Surf A. Physicochem Eng Asp 2020; 601: 124999.
43. Rathod S, Arya S, Shukla R, Ray D, Aswal VK, Bahadur P, et al. Investigations on the role of edge activator upon structural transitions in Span vesicles. Colloids Surf A. Physicochem Eng Asp 2021; 627: 127246.
44. Florence AT, Attwood D. Physicochemical Principles of Pharmacy: In Manufacture, Formulation and Clinical Use. Pharmaceutical press 2015. p.664.
45. Samimi S, Maghsoudnia N, Eftekhari RB, Dorkoosh F. Lipid-based nanoparticles for drug delivery systems. Charact Biol Nanomater Drug Delivery 2019; 47-76.
46. Shao J, Liang R, Ding D, Zheng X, Zhu X, Hu S, et al. A smart multifunctional nanoparticle for enhanced near-infrared image-guided photothermal therapy against gastric cancer. Int J Nanomedicine 2021; 16: 2897-2915.
47. Ting CW, Chou YH, Huang SY, Chiang WH. Indocyanine green-carrying polymeric nanoparticles with acid-triggered detachable PEG coating and drug release for boosting cancer photothermal therapy. Colloids Surf B Biointerfaces 2021; 208: 112048.
48. Wang YG, Kim H, Mun S, Kim D, Choi Y. Indocyanine green-loaded perfluorocarbon nanoemulsions for bimodal (19)F-magnetic resonance/nearinfrared fluorescence imaging and subsequent phototherapy. Quant Imaging Med Surg 2013; 3: 132-140.
49. Bilici K, Muti A, Sennaroğlu A, Yagci Acar H. Indocyanine green loaded APTMS coated SPIONs for dual phototherapy of cancer. J Photochem Photobiol B 2019; 201: 111648.
50. Ma Y, Liu X, Ma Q, Liu Y. Near-infrared nanoparticles based on indocyanine green-conjugated albumin: A versatile platform for imaging-guided synergistic tumor chemo-phototherapy with temperature-responsive drug release. Onco Targets Ther 2018; 11: 8517-8528.
51. Hao Y, Chung CK, Gu Z, Schomann T, Dong X, Veld R, et al. Combinatorial therapeutic approaches of photodynamic therapy and immune checkpoint blockade for colon cancer treatment. Mol Biomed 2022; 3: 26-45.
52. Vital-Fujii DG, Baptista MS. Progress in the photodynamic therapy treatment of Leishmaniasis. Braz J Med Biol Res 2021; 54: e11570.
53. Huang YY, Chen AC, Carroll JD, Hamblin MR. Biphasic dose response in low level light therapy. Dose Response 2009; 7: 358-383.
54. Cabral FV, Souza THDS, Sellera FP, Fontes A, Ribeiro MS. Towards effective cutaneous leishmaniasis treatment with light-based technologies. A systematic review and meta-analysis of preclinical studies. J Photochem Photobiol B 2021; 221:112236
55. Shirata C, Kaneko J, Inagaki Y, Kokudo T, Sato M, Kiritani S, et al. Near-infrared photothermal/photodynamic therapy with indocyanine green induces apoptosis of hepatocellular carcinoma cells through oxidative stress. Sci Rep 2017; 7: 13958-13965.
56. Long S, Xu Y, Zhou F, Wang B, Yang Y, Fu Y, et al. Characteristics of temperature changes in photothermal therapy induced by combined application of indocyanine green and laser. Oncol Lett 2019; 17: 3952-3959.