LED Light Source for in vitro Study of Photosensitizing Agents for Photodynamic Therapy

The aim of the investigation was to develop a LED light source providing a homogeneous light distribution in 96-well plates and allowing an independent irradiation of individual wells, as well as its experimental testing in in vitro study of photosensitizers for photodynamic therapy.

Materials and Methods. The experiments were carried out on human cell lines of epidermoid carcinoma А-431 and human bladder carcinoma Т24. Two photosensitizers for fluorescence diagnostics and photodynamic therapy were used: Photosens®, and newly synthesized polymer brushes nanoparticles doped with porphyrazine chromophore. To study the photo-activity of the agents we created a light source with replaceable LED-arrays. Photodynamic activity of the photosensitizers was estimated in vitro by MTT assay.

Results. The created LED light source enables to expose cell cultures with narrow-band irradiation with different wavelengths and radiation intensity being up to 90 mW/cm². Precision control of temperature conditions during the investigation is provided. Light power instability is less than 1%. Independent on and off switching of LED clusters of 4 elements is provided in order to illuminate several wells groups of a standard 96-well culture plate with different light doses simultaneously. The developed light source was tested in the study of photo-activity of agents for photodynamic therapy. The dependence of Photosens® toxicity on the light dose was evaluated and significant photodynamic activity of newly synthesized porphyrazine fluorophore was demonstrated.

Conclusion. A newly developed LED light source with replaceable LED-arrays with narrow spectral bands provides an effective in vitro study of photosensitizing preparations under development. The state-of-the-art approach enables high throughput screening of promising agents for fluorescence diagnostics and photodynamic therapy.

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Shilyagina N.Y., Plekhanov V.I., Shkunov I.V., Shilyagin P.А., Dubasova L.V., Brilkina А.А., Sokolova E.A., Turchin I.V., Balalaeva I.V. LED Light Source for in vitro Study of Photosensitizing Agents for Photodynamic Therapy. Sovremennye tehnologii v medicine 2014; 6(2): 15