In vitro и in vivo photodynamic therapy of solid tumors with a combination of riboflavin and upconversion nanoparticles

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Abstract

Rationale: Riboflavin (vitamin B2) is one of the most promising agents for photodynamic therapy (PDT). However, its use is limited by the excitation in the ultraviolet (UV) and visible spectral ranges and, as a result, by a small penetration into biological tissue not exceeding a few millimeters. This problem could be solved by approaches ensuring excitation of riboflavin molecules within tumor tissues by infrared (IR) light. Upconversion nanoparticles (UCNPs) can be potentially considered as mediators able to effectively convert the exciting radiation of the near IR range, penetrating into biological tissue to a 3 cm depth, into the photoluminescence in the UV and visible spectral ranges.

Aim: To evaluate the efficacy of UCNPs for IR-mediated riboflavin activation in the depth of tumor tissue during PDT.
Materials and methods: The water-soluble riboflavin flavin mononucleotide (FMN, Pharmstandard-UfaVITA, Russia) was used as a photosensitizer in in vitro and in vivo experiments. The in vitro experiments were performed on human breast adenocarcinoma SK-BR-3, human glioblastoma U-87 MG, and rat glioma C6 cell lines. Lewis lung carcinoma (LLC) inoculated to hybrid BDF1 mice was used as a model to demonstrate the delivery of FMN to the tumor. UCNPs with a core/shell structure [NaYF4:Yb3+, Tm3+/NaYF4] were used for photoactivation of FMN in vivo. PDT based on FMN, UCNPs and laser radiation 975 nm (IR) was performed on mouse xenografts of human breast adenocarcinoma SKBR-3.

Results: We were able to show that FMN could act as an effective in vitro photosensitizer for SK-BR-3, U-87 MG, and C6 cell lines. FMN IC50 values for glioma cells were ~30 μM, and for SK-BR-3 cell line ~50 μM (24 h incubation, irradiation 4.2 J/cm2). In the LLC model, the appropriate concentration of FMN (30 μM and above) can be achieved in the tumor as a result of systemic administration of FMN (at 2 and 24 hours after injection). The effect of PDT using near IR light for UCNP-mediated excitation of FMN was demonstrated in mouse xenografts SKBR-3, with the tumor growth inhibition of 90±5%.

Conclusion: The study has demonstrated the possibility to use riboflavin (vitamin B2) as a photosensitizer for PDT. The photoexcitation of FMN via the anti-Stokes photoluminescence of UCNPs allows for implementation of the PDT technique with the near IR spectral range.

About the authors

N. V. Sholina

N.N. Blokhin National Medical Research Centre of Oncology; I.M. Sechenov First Moscow State Medical University; Federal Scientific Research Centre Crystallography and Photonics, Russian Academy of Sciences

Author for correspondence.
Email: SholinaNV@gmail.com
ORCID iD: 0000-0001-9866-2878

Natal'ya V. Sholina - Postgraduate Student, Junior Research Fellow, Laboratory of Biomarkers and Mechanisms of Tumor Angiogenesis N.N. Blokhin NMRCO; Laboratory of Photon Bioengineering, Institute of Molecular Medicine I.M. Sechenov FMSMU; Laboratory of Laser Biomedicine FSRC"Crystallography and Photonic, RAS.

24 Kashirskoe shosse, Moscow, 115478; 8/2 Trubetskaya ul., Moscow, 119991; 59 Leninsky prospekt, Moscow, 119333, tel.: + 7 (926) 957 96 63

Russian Federation

R. A. Akasov

I.M. Sechenov First Moscow State Medical University; Federal Scientific Research Centre Crystallography and Photonics, Russian Academy of Sciences

Email: fake@neicon.ru
ORCID iD: 0000-0001-6486-8114

Roman A. Akasov - PhD, Research Fellow, Laboratory of Photon Bioengineering, Institute of Molecular Medicine I.M. Sechenov FMSMU; Laboratory of Laser Biomedicine FSRCCrystallography and Photonic, RAS.

8/2 Trubetskaya ul., Moscow, 119991; 59 Leninsky prospekt, Moscow, 119333

Russian Federation

D. A. Khochenkov

N.N. Blokhin National Medical Research Centre of Oncology

Email: fake@neicon.ru
ORCID iD: 0000-0002-5694-3492

Dmitry A. Khochenkov - PhD (in Biol.), Head of the Laboratory of Biomarkers and Mechanisms of Tumor Angiogenesis N.N. Blokhin NMRCO.

24 Kashirskoe shosse, Moscow, 115478

Russian Federation

A. N. Generalova

Federal Scientific Research Centre Crystallography and Photonics, Russian Academy of Sciences; M.M. Shemyakin – Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: fake@neicon.ru
ORCID iD: 0000-0001-9646-1693

Alla N. Generalova - PhD (in Chem.), Senior Research Fellow, Laboratory of Laser Biomedicine FSRCCrystallography and Photonic, RAS; Laboratory "Polymers for Biology" M.M. Shemyakin - Yu.A. Ovchinnikov IBC, RAS.

59 Leninsky prospekt, Moscow, 119333; 16/10 Miklukho-Maklaya ul., Moscow, 117997

Russian Federation

V. A. Semchishen

Federal Scientific Research Centre Crystallography and Photonics, Russian Academy of Sciences

Email: fake@neicon.ru
ORCID iD: 0000-0003-1035-3013

Vladimir A. Semchishen – PhD (in Phys. and Math.), Leading Research Fellow, Laboratory of Laser Biomedicine.

59 Leninsky prospekt, Moscow, 119333

Russian Federation

E. V. Khaydukov

I.M. Sechenov First Moscow State Medical University; Federal Scientific Research Centre Crystallography and Photonics, Russian Academy of Sciences

Email: fake@neicon.ru
ORCID iD: 0000-0002-3900-2949

Evgeny V. Khaydukov - PhD (in Phys. and Math.), Head of the Laboratory of Photon Bioengineering, Institute of Molecular Medicine I.M. Sechenov FMSMU; Head of the Laboratory of Laser Biomedicine FSRC"Crystallography and Photonic, RAS.

8/2 Trubetskaya ul., Moscow, 119991; 59 Leninsky prospekt, Moscow, 119333

Russian Federation

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Copyright (c) 2019 Sholina N.V., Akasov R.A., Khochenkov D.A., Generalova A.N., Semchishen V.A., Khaydukov E.V.

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