Synergistic effect of cold atmospheric plasma and methylene blue loaded nano micelles on treating human glioblastoma cells: An in vitro and molecular dynamics study

Document Type : Original Article

Authors

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

2 Department of Medical Physics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

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

10.22038/ijbms.2024.79858.17304

Abstract

Objective(s): One of the most recent cancer treatment methods is cold atmospheric plasma (CAP), which destroys cancer cells without affecting healthy cells. Also, the created photons in the CAP flame can be used to excite a proper photosensitizing agent (PS). Therefore, using nano micelle systems containing a proper photosensitizer may be beneficial in raising the treatment efficacy of CAP. In this study, we utilized molecular dynamics (MD) simulation to optimize a nano micellar system containing methylene blue to take advantage of the induced photodynamic effect of a CAP generator with helium gas on a glioblastoma cell line.
Materials and Methods: Some micelle properties were first determined and optimized by MD with GROMACS software. Then, micelles containing methylene blue (Micelle-MB) and free methylene blue (MB) at various concentrations were prepared. Singlet oxygen dosimetry using 1,3-diphenylisobenzofuran(DPBF)was performed in the presence and absence of Micelle-MB and MB. Subsequently, the cytotoxicity of MB and Micelle-MB was evaluated on U87-MG cancer cells, and their half-maximal inhibitory concentrations (IC50) were determined. After 48 hr of treatment, the percentage of cell survival was determined using the MTT test. The experiments were repeated at least three times. The synergy index was selected to compare the results.
Results: Treatment with CAP and MB reduced the survival rate compared to the PS-free group with CAP. Results of singlet oxygen dosimetry showed that Micelle-MB might be more efficient in producing ROS. CAP treatment with Micelle-MB resulted in more cell death than free MB. In addition, cell viability decreased in Micelle-MB groups with increasing irradiation time in the three investigated irradiation times.
Conclusion: Using Micelle-MB in the CAP treatment improves treatment efficiency in the U87-MG cell line.

Keywords

Main Subjects

References

1. Cooper GM, Adams K. The cell: A molecular approach: Oxford University Press; 2022.
2. Czarnywojtek A, Borowska M, Dyrka K, Van Gool S, Sawicka-Gutaj N, Moskal J, et al. Glioblastoma multiforme: the latest diagnostics and treatment techniques. Pharmacology 2023; 108:423-431.
3. Ihata T, Nonoguchi N, Fujishiro T, Omura N, Kawabata S, Kajimoto Y, et al. The effect of hypoxia on photodynamic therapy with 5-aminolevulinic acid in malignant gliomas. Photodiagnosis Photodyn Ther 2022; 40:103056.
4. Whelan HT. High-grade glioma/glioblastoma multiforme: Is there a role for photodynamic therapy? J Natl Compr Canc Netw 2012; 10:S-31-S-34.
5. Quirk BJ, Brandal G, Donlon S, Vera JC, Mang TS, Foy AB, et al. Photodynamic therapy (PDT) for malignant brain tumors–where do we stand? Photodiagnosis Photodyn Ther 2015; 12:530-544.
6. Pucelik B, Sułek A, Barzowska A, Dąbrowski JM. Recent advances in strategies for overcoming hypoxia in photodynamic therapy of cancer. Cancer Lett 2020; 492:116-135.
7. Noghreiyan AV, Imanparast A, Ara ES, Soudmand S, Noghreiyan VV, Sazgarnia A. In vitro investigation of cold atmospheric plasma induced photodynamic effect by Indocyanine green and Protoporphyrin IX. Photodiagnosis Photodyn Ther 2020; 31:101822.
8. Ara ES, Noghreiyan AV, Sazgarnia A. Evaluation of photodynamic effect of Indocyanine green (ICG) on the colon and glioblastoma cancer cell lines pretreated by cold atmospheric plasma. Photodiagnosis Photodyn Ther 2021; 35:102408.
9. Momeni S, Shanei A, Sazgarnia A, Attaran N, Aledavood SA. The synergistic effect of cold atmospheric plasma mediated gold nanoparticles conjugated with Indocyanine green as an innovative approach to cooperation with radiotherapy. Cell J 2023; 25:51-61.
10. Privat-Maldonado A, Gorbanev Y, Dewilde S, Smits E, Bogaerts A. Reduction of human glioblastoma spheroids using cold atmospheric plasma: The combined effect of short-and long-lived reactive species. Cancers 2018; 10:394-410.
11. Faramarzi F, Zafari P, Alimohammadi M, Moonesi M, Rafiei A, Bekeschus S. Cold physical plasma in cancer therapy: mechanisms, signaling, and immunity. Oxid Med Cell Longev 2021; 2021:9916796.
12. Tardivo JP, Del Giglio A, De Oliveira CS, Gabrielli DS, Junqueira HC, Tada DB, et al. Methylene blue in photodynamic therapy: From basic mechanisms to clinical applications. Photodiagnosis Photodyn Ther 2005; 2:175-191.
13. Wu P-T, Lin C-L, Lin C-W, Chang N-C, Tsai W-B, Yu J. Methylene-blue-encapsulated liposomes as photodynamic therapy nano agents for breast cancer cells. Nanomaterials 2018; 9:14-25.
14. Bharate GY, Qin H, Fang J. Poly (styrene-co-maleic acid) micelle of photosensitizers for targeted photodynamic therapy, exhibits prolonged singlet oxygen generating capacity and superior intracellular uptake. J Pers Med 2022; 12:493-512.
15. Joseph M, Trinh HM, Mitra AK. Peptide and protein-based therapeutic agents.  Emerging nanotechnologies for diagnostics,  drug delivery and medical devices: Elsevier; 2017. p. 145-167.
16.    Koopaie M. Nanoparticulate systems for dental drug delivery.  Nanoengineered Biomaterials for Advanced Drug Delivery: Elsevier; 2020. p. 525-559.
17. Perumal S, Atchudan R, Lee W. A review of polymeric micelles and their applications. Polymers  2022; 14:2510.
18. Mobasheri M, Attar H, Rezayat Sorkhabadi SM, Khamesipour A, Jaafari MR. Solubilization behavior of polyene antibiotics in nanomicellar system: insights from molecular dynamics simulation of the amphotericin B and nystatin interactions with polysorbate 80. Molecules 2015; 21:6-31.
19. Periole X, Marrink S-J. The Martini coarse-grained force field. Methods Mol Biol 2013:533-565.
20. Marrink SJ, Risselada HJ, Yefimov S, Tieleman DP, De Vries AH. The MARTINI force field: coarse grained model for biomolecular simulations. J Phys Chem B 2007; 111:7812-7824.
21. 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.
22. Alargova RG, Kochijashky II, Sierra ML, Zana R. Micelle Aggregation Numbers of Surfactants in Aqueous Solutions:  A Comparison between the Results from Steady-State and Time-Resolved Fluorescence Quenching. Langmuir 1998; 14:5412-5418.
23. Jesus VPS, Raniero L, Lemes GM, Bhattacharjee TT, Caetano Júnior PC, Castilho ML. Nanoparticles of methylene blue enhance photodynamic therapy. Photodiagnosis Photodyn Ther 2018; 23:212-217.
24. Patravale VB, Upadhaya PG, Jain RD. Preparation and characterization of micelles. Methods Mol Biol 2019; 2000:19-29.
25.    Soares JCM, Luiz MT, Oshiro Junior JA, Besegato JF, de Melo PBG, Rastelli ANS, et al. Antimicrobial photodynamic therapy mediated by methylene blue-loaded polymeric micelles against Streptococcus mutans and Candida albicans biofilms. Photodiagnosis Photodyn Ther 2023; 41:103285.
26. Lv G, Wang F, Cai W, Zhang X. Characterization of the addition of lipophilic Span 80 to the hydrophilic Tween 80-stabilized emulsions. Colloids Surf A Physicochem Eng Asp 2014; 447:8-13.
27. Owen SC, Chan DPY, Shoichet MS. Polymeric micelle stability. Nano Today 2012; 7:53-65.
28. Yan L, Miller J, Yuan M, Liu JF, Busch TM, Tsourkas A, et al. Improved photodynamic therapy efficacy of protoporphyrin IX-Loaded polymeric micelles using erlotinib pretreatment. Biomacromolecules 2017; 18:1836-1844.
29. Umezawa M, Kobayashi H, Ichihashi K, Sekiyama S, Okubo K, Kamimura M, et al. Heat treatment effects for controlling dye molecular states in the hydrophobic core of over-1000 nm near-infrared (NIR-II) fluorescent micellar nanoparticles. ACS Omega 2022; 7:5817-5824.
30.    Tian Z, Li H, Liu Z, Yang L, Zhang C, He J, et al. Enhanced photodynamic therapy by improved light energy capture efficiency of porphyrin photosensitizers. Curr Treat Options Oncol 2023; 24:1274-1292.
31. Hari SK, Gauba A, Shrivastava N, Tripathi RM, Jain SK, Pandey AK. Polymeric micelles and cancer therapy: An ingenious multimodal tumor-targeted drug delivery system. Drug Deliv Transl Res 2023; 13:135-163.
32. Tavakkoli Yaraki M, Liu B, Tan YN. Emerging strategies in enhancing singlet oxygen generation of nano-photosensitizers toward advanced phototherapy. Nanomicro Lett 2022; 14:123-171.
33. Shanmugham V, Subban R. Capsanthin-loaded micelles: Preparation, characterization and in vitro evaluation of cytotoxicity using MDA-MB-231 breast cancer cell line. Food Technol Biotechnol 2022; 60:350-360.
34. Kalghatgi S, Friedman G, Fridman A, Clyne AM. Endothelial cell proliferation is enhanced by low dose non-thermal plasma through fibroblast growth factor-2 release. Ann Biomed Eng 2010; 38:748-757.
35. Moniruzzaman R, Rehman MU, Zhao Q-L, Jawaid P, Takeda K, Ishikawa K, et al. Cold atmospheric helium plasma causes synergistic enhancement in cell death with hyperthermia and an additive enhancement with radiation. Sci Rep 2017; 7:11659.
36. Tang W, Xu H, Kopelman R, Philbert MA. Photodynamic characterization and in vitro application of methylene blue‐containing nanoparticle platforms. Photochem Photobiol 2005; 81:242-249.
37. Tu C, Zhu L, Li P, Chen Y, Su Y, Yan D, et al. Supramolecular polymeric micelles by the host–guest interaction of star-like calix [4] arene and chlorin e6 for photodynamic therapy. Chem Commun 2011; 47:6063-6065.
38. Abrahamse H, Hamblin MR. New photosensitizers for photodynamic therapy. Biochem J 2016; 473:347-364.
39. Lopez-Quintela MA. Synthesis of nanomaterials in microemulsions: formation mechanisms and growth control. Curr Opin Colloid Interface Sci 2003; 8:137-144.
40. Junqueira HC, Severino D, Dias LG, Gugliotti MS, Baptista MS. Modulation of methylene blue photochemical properties based on adsorption at aqueous micelle interfaces. Physical Chemistry Chemical Physics 2002; 4:2320-2328.
41. Alemayehu YA, Fan WL, Ilhami FB, Chiu C-W, Lee DJ, Cheng CC. Photosensitive supramolecular micelle-mediated cellular uptake of anticancer drugs enhances the efficiency of chemotherapy. Int J  Mol Sci 2020; 21:4677.
42. Zhou W, Li C, Wang Z, Zhang W, Liu J. Factors affecting the stability of drug-loaded polymeric micelles and strategies for improvement. J Nanopart Res 2016; 18:1-18.
Iranian Journal of Basic Medical Sciences
Volume 28, Issue 3
March 2025
Pages 299-309

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