Limits...
Survival analysis of F98 glioma rat cells following minibeam or broad-beam synchrotron radiation therapy.

Gil S, Sarun S, Biete A, Prezado Y, Sabés M - Radiat Oncol (2011)

Bottom Line: A dose escalation study was performed in order to delimit the range of doses where a therapeutic effect could be expected.These results will help in the design and optimization of the forthcoming in vivo studies at the ESRF.In addition, flow cytometric analysis pointed at a larger effectiveness for minibeams, due to the higher proportion of early apoptotic cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centre d'Estudis en Biofísica, Faculty of Medicine, Autonomous University of Barcelona, Cerdanyola del Vallès, Spain. silvia.gild@campus.uab.cat

ABSTRACT

Background: In the quest of a curative radiotherapy treatment for gliomas new delivery modes are being explored. At the Biomedical Beamline of the European Synchrotron Radiation Facility (ESRF), a new spatially-fractionated technique, called Minibeam Radiation Therapy (MBRT) is under development. The aim of this work is to compare the effectiveness of MBRT and broad-beam (BB) synchrotron radiation to treat F98 glioma rat cells. A dose escalation study was performed in order to delimit the range of doses where a therapeutic effect could be expected. These results will help in the design and optimization of the forthcoming in vivo studies at the ESRF.

Methods: Two hundred thousand F98 cells were seeded per well in 24-well plates, and incubated for 48 hours before being irradiated with spatially fractionated and seamless synchrotron x-rays at several doses. The percentage of each cell population (alive, early apoptotic and dead cells, where either late apoptotic as necrotic cells are included) was assessed by flow cytometry 48 hours after irradiation, whereas the metabolic activity of surviving cells was analyzed on days 3, 4, and 9 post-irradiation by using QBlue test.

Results: The endpoint (or threshold dose from which an important enhancement in the effectiveness of both radiation treatments is achieved) obtained by flow cytometry could be established just before 12 Gy in the two irradiation schemes, whilst the endpoints assessed by the QBlue reagent, taking into account the cell recovery, were set around 18 Gy in both cases. In addition, flow cytometric analysis pointed at a larger effectiveness for minibeams, due to the higher proportion of early apoptotic cells.

Conclusions: When the valley doses in MBRT equal the dose deposited in the BB scheme, similar cell survival ratio and cell recovery were observed. However, a significant increase in the number of early apoptotic cells were found 48 hours after the minibeam radiation in comparison with the seamless mode.

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24-well plate's picture (partially). Picture corresponding to a 24-well plate like those employed in the irradiations, either for MBRT or seamless irradiations. The verification of the dose assessed was done by using radiochromic films just above the wells as it is shown.
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Figure 1: 24-well plate's picture (partially). Picture corresponding to a 24-well plate like those employed in the irradiations, either for MBRT or seamless irradiations. The verification of the dose assessed was done by using radiochromic films just above the wells as it is shown.

Mentions: The dose deposited on the cells monolayer was assessed both theoretically (Monte Carlo simulations) and experimentally. In order to reduce an extra dose deposition on cells from the scattering of filled wells contiguously, the seeding well configuration for each plate was the same as those represented in Figure 1. Moreover, to have similar valley doses like the ones expected in the forthcoming in vivo experiments with small animals the wells were filled up with medium, remaining the cells on the bottom, at 1.7 cm depth. If the cell layer had been irradiated directly, in the absence of medium, the peak-to-valley dose ratio (PVDR) would have been much higher than the one calculated for the in vivo experiments with rats, due to reduced scattering filling the valleys [4].


Survival analysis of F98 glioma rat cells following minibeam or broad-beam synchrotron radiation therapy.

Gil S, Sarun S, Biete A, Prezado Y, Sabés M - Radiat Oncol (2011)

24-well plate's picture (partially). Picture corresponding to a 24-well plate like those employed in the irradiations, either for MBRT or seamless irradiations. The verification of the dose assessed was done by using radiochromic films just above the wells as it is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3094367&req=5

Figure 1: 24-well plate's picture (partially). Picture corresponding to a 24-well plate like those employed in the irradiations, either for MBRT or seamless irradiations. The verification of the dose assessed was done by using radiochromic films just above the wells as it is shown.
Mentions: The dose deposited on the cells monolayer was assessed both theoretically (Monte Carlo simulations) and experimentally. In order to reduce an extra dose deposition on cells from the scattering of filled wells contiguously, the seeding well configuration for each plate was the same as those represented in Figure 1. Moreover, to have similar valley doses like the ones expected in the forthcoming in vivo experiments with small animals the wells were filled up with medium, remaining the cells on the bottom, at 1.7 cm depth. If the cell layer had been irradiated directly, in the absence of medium, the peak-to-valley dose ratio (PVDR) would have been much higher than the one calculated for the in vivo experiments with rats, due to reduced scattering filling the valleys [4].

Bottom Line: A dose escalation study was performed in order to delimit the range of doses where a therapeutic effect could be expected.These results will help in the design and optimization of the forthcoming in vivo studies at the ESRF.In addition, flow cytometric analysis pointed at a larger effectiveness for minibeams, due to the higher proportion of early apoptotic cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centre d'Estudis en Biofísica, Faculty of Medicine, Autonomous University of Barcelona, Cerdanyola del Vallès, Spain. silvia.gild@campus.uab.cat

ABSTRACT

Background: In the quest of a curative radiotherapy treatment for gliomas new delivery modes are being explored. At the Biomedical Beamline of the European Synchrotron Radiation Facility (ESRF), a new spatially-fractionated technique, called Minibeam Radiation Therapy (MBRT) is under development. The aim of this work is to compare the effectiveness of MBRT and broad-beam (BB) synchrotron radiation to treat F98 glioma rat cells. A dose escalation study was performed in order to delimit the range of doses where a therapeutic effect could be expected. These results will help in the design and optimization of the forthcoming in vivo studies at the ESRF.

Methods: Two hundred thousand F98 cells were seeded per well in 24-well plates, and incubated for 48 hours before being irradiated with spatially fractionated and seamless synchrotron x-rays at several doses. The percentage of each cell population (alive, early apoptotic and dead cells, where either late apoptotic as necrotic cells are included) was assessed by flow cytometry 48 hours after irradiation, whereas the metabolic activity of surviving cells was analyzed on days 3, 4, and 9 post-irradiation by using QBlue test.

Results: The endpoint (or threshold dose from which an important enhancement in the effectiveness of both radiation treatments is achieved) obtained by flow cytometry could be established just before 12 Gy in the two irradiation schemes, whilst the endpoints assessed by the QBlue reagent, taking into account the cell recovery, were set around 18 Gy in both cases. In addition, flow cytometric analysis pointed at a larger effectiveness for minibeams, due to the higher proportion of early apoptotic cells.

Conclusions: When the valley doses in MBRT equal the dose deposited in the BB scheme, similar cell survival ratio and cell recovery were observed. However, a significant increase in the number of early apoptotic cells were found 48 hours after the minibeam radiation in comparison with the seamless mode.

Show MeSH
Related in: MedlinePlus