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Screening of two-photon activated photodynamic therapy sensitizers using a 3D osteosarcoma model

(2019) ANALYST. 144(9). p.3056-3063
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Abstract
Photodynamic therapy (PDT) involves a photosensitizing agent activated with light to induce cell death. Two-photon excited PDT (TPE-PDT) offers numerous benefits compared to traditional one-photon induced PDT, including an increased penetration depth and precision. However, the in vitro profiling and comparison of two-photon photosensitizers (PS) are still troublesome. Herein, we report the development of an in vitro screening platform of TPE-PS using a 3D osteosarcoma cell culture. The platform was tested using three different two-photon (2P) active compounds - a 2P sensitizer P2CK, a fluorescent dye Eosin Y, and a porphyrin derivative (TPP). Their 2P absorption cross-sections (sigma(2PA)) were characterised using a fully automated z-scan setup. TPP exhibited a remarkably high sigma(2PA) at 720 nm (8865 GM) and P2CK presented a high absorption at 850 nm (405 GM), while Eosin Y had the lowest 2P absorption at the studied wavelengths (<100 GM). The cellular uptake of PS visualized using confocal laser scanning microscopy showed that both TPP and P2CK were internalized by the cells, while Eosin Y stayed mainly in the surrounding media. The efficiency of the former two TPE-PS was quantified using the PrestoBlue metabolic assay, showing a significant reduction in cell viability after two-photon irradiation. The possibility of damage localization was demonstrated using a co-culture of adipose derived stem cells together with osteosarcoma spheroids showing no signs of damage to the surrounding healthy cells after TPE-PDT.
Keywords
SINGLET OXYGEN, PHOTOSENSITIZERS, EXCITATION, CELLS, PENETRATION

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MLA
Dobos, Agnes, et al. “Screening of Two-Photon Activated Photodynamic Therapy Sensitizers Using a 3D Osteosarcoma Model.” ANALYST, vol. 144, no. 9, 2019, pp. 3056–63.
APA
Dobos, A., Steiger, W., Theiner, D., Gruber, P., Lunzer, M., Van Hoorick, J., … Ovsianikov, A. (2019). Screening of two-photon activated photodynamic therapy sensitizers using a 3D osteosarcoma model. ANALYST, 144(9), 3056–3063.
Chicago author-date
Dobos, Agnes, Wolfgang Steiger, Dominik Theiner, Peter Gruber, Markus Lunzer, Jasper Van Hoorick, Sandra Van Vlierberghe, and Aleksandr Ovsianikov. 2019. “Screening of Two-Photon Activated Photodynamic Therapy Sensitizers Using a 3D Osteosarcoma Model.” ANALYST 144 (9): 3056–63.
Chicago author-date (all authors)
Dobos, Agnes, Wolfgang Steiger, Dominik Theiner, Peter Gruber, Markus Lunzer, Jasper Van Hoorick, Sandra Van Vlierberghe, and Aleksandr Ovsianikov. 2019. “Screening of Two-Photon Activated Photodynamic Therapy Sensitizers Using a 3D Osteosarcoma Model.” ANALYST 144 (9): 3056–3063.
Vancouver
1.
Dobos A, Steiger W, Theiner D, Gruber P, Lunzer M, Van Hoorick J, et al. Screening of two-photon activated photodynamic therapy sensitizers using a 3D osteosarcoma model. ANALYST. 2019;144(9):3056–63.
IEEE
[1]
A. Dobos et al., “Screening of two-photon activated photodynamic therapy sensitizers using a 3D osteosarcoma model,” ANALYST, vol. 144, no. 9, pp. 3056–3063, 2019.
@article{8610535,
  abstract     = {Photodynamic therapy (PDT) involves a photosensitizing agent activated with light to induce cell death. Two-photon excited PDT (TPE-PDT) offers numerous benefits compared to traditional one-photon induced PDT, including an increased penetration depth and precision. However, the in vitro profiling and comparison of two-photon photosensitizers (PS) are still troublesome. Herein, we report the development of an in vitro screening platform of TPE-PS using a 3D osteosarcoma cell culture. The platform was tested using three different two-photon (2P) active compounds - a 2P sensitizer P2CK, a fluorescent dye Eosin Y, and a porphyrin derivative (TPP). Their 2P absorption cross-sections (sigma(2PA)) were characterised using a fully automated z-scan setup. TPP exhibited a remarkably high sigma(2PA) at 720 nm (8865 GM) and P2CK presented a high absorption at 850 nm (405 GM), while Eosin Y had the lowest 2P absorption at the studied wavelengths (<100 GM). The cellular uptake of PS visualized using confocal laser scanning microscopy showed that both TPP and P2CK were internalized by the cells, while Eosin Y stayed mainly in the surrounding media. The efficiency of the former two TPE-PS was quantified using the PrestoBlue metabolic assay, showing a significant reduction in cell viability after two-photon irradiation. The possibility of damage localization was demonstrated using a co-culture of adipose derived stem cells together with osteosarcoma spheroids showing no signs of damage to the surrounding healthy cells after TPE-PDT.},
  author       = {Dobos, Agnes and Steiger, Wolfgang and Theiner, Dominik and Gruber, Peter and Lunzer, Markus and Van Hoorick, Jasper and Van Vlierberghe, Sandra and Ovsianikov, Aleksandr},
  issn         = {0003-2654},
  journal      = {ANALYST},
  keywords     = {SINGLET OXYGEN,PHOTOSENSITIZERS,EXCITATION,CELLS,PENETRATION},
  language     = {eng},
  number       = {9},
  pages        = {3056--3063},
  title        = {Screening of two-photon activated photodynamic therapy sensitizers using a 3D osteosarcoma model},
  url          = {http://dx.doi.org/10.1039/c9an00068b},
  volume       = {144},
  year         = {2019},
}

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