Issue 9, 2019

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

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 (σ2PA) were characterised using a fully automated z-scan setup. TPP exhibited a remarkably high σ2PA 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.

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

Supplementary files

Article information

Article type
Paper
Submitted
11 янв 2019
Accepted
06 мар 2019
First published
27 мар 2019
This article is Open Access
Creative Commons BY license

Analyst, 2019,144, 3056-3063

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

A. Dobos, W. Steiger, D. Theiner, P. Gruber, M. Lunzer, J. Van Hoorick, S. Van Vlierberghe and A. Ovsianikov, Analyst, 2019, 144, 3056 DOI: 10.1039/C9AN00068B

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