Abstract
Temporal dependence of changes in the morphological characteristics of cells of two cultured lines of cancer origin, HeLa and A549, induced by photodynamic treatment with Radachlorin photosensitizer, have been monitored using digital holographic microscopy during first two hours after short-term irradiation. The observed post-treatment early dynamics of the phase shift in the transmitted wavefront indicated several distinct scenarios of cell behavior depending upon the irradiation dose. In particular the phase shift increased at low doses, which can be associated with apoptosis, while at high doses it decreased, which can be associated with necrosis. As shown, the two cell types responded differently to similar irradiation doses. Although the sequence of death scenarios with the increase of the irradiation dose was the same, each scenario was realized at substantially different doses. These findings suggest that the average phase shift of the transmitted wavefront can be used for quantitative non-invasive cell death characterization. The conclusions made were cofirmed by commonly used test assays using confocal fluorescent microscopy.
Highlights
A persistent increase of cancer incidences and high recurrence rate are key problems in modern oncology requiring improvement of existing treat-Preprint submitted to Elsevier ment modalities and development of novel approaches for early diagnostics and therapy
In [18] we presented our first results on determination of cellular morphology at PD treatment by means of digital holographic microscopy
In this paper we present results on comprehensive monitoring of variations in morphological characteristics of the two widely used cultured cancer cell lines in the course and after PD treatment with chlorin PS at various irradiation doses
Summary
A persistent increase of cancer incidences and high recurrence rate are key problems in modern oncology requiring improvement of existing treat-Preprint submitted to Elsevier ment modalities and development of novel approaches for early diagnostics and therapy. One of highly promising modalities is photodynamic therapy (PDT) which was already successfully applied for treatment of various malignant and benign pathologies, skin lesions, macular degeneration, microbial infections etc. The anti-tumor effect of PDT is provided by the three interconnected processes: direct death of tumor cells, vascular disruption and activation of immune response [4]. The integral effect of PDT is known relatively well, contributions and mechanisms of occurring processes are still far from being sufficiently understood [5, 6]. It was presumed that the contribution of each of these processes to the tumor response to PDT may vary depending upon the PS type and PDT protocol parameters, irradiation dose and duration in particular. The analysis of individual cells’ response to photodynamic (PD) treatment at various conditions is an essential aim for the study of PS efficacy as a prerequisite for evaluation of optimal treatment doses
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