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                <title level="j">Radiation Oncology</title>
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                  <date type="datePub">2022-12</date>
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                <term xml:lang="en">DNA repair</term>
                <term xml:lang="en">Hypoxia</term>
                <term xml:lang="en">Laser-driven protons</term>
                <term xml:lang="en">Ultra-high dose rate</term>
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              <classCode scheme="halDomain" n="phys.phys.phys-plasm-ph">Physics [physics]/Physics [physics]/Plasma Physics [physics.plasm-ph]</classCode>
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              <p>Background: There is currently signifcant interest in assessing the role of oxygen in the radiobiological efects atultra-high dose rates. Oxygen modulation is postulated to play a role in the enhanced sparing efect observed inFLASH radiotherapy, where particles are delivered at 40–1000 Gy/s. Furthermore, the development of laser-drivenaccelerators now enables radiobiology experiments in extreme regimes where dose rates can exceed 109 Gy/s, andpredicted oxygen depletion efects on cellular response can be tested. Access to appropriate experimental enviroments, allowing measurements under controlled oxygenation conditions, is a key requirement for these studies.We report on the development and application of a bespoke portable hypoxia chamber specifcally designed forexperiments employing laser-driven sources, but also suitable for comparator studies under FLASH and conventionalirradiation conditions.Materials and methods: We used oxygen concentration measurements to test the induction of hypoxia and themaintenance capacity of the chambers. Cellular hypoxia induction was verifed using hypoxia inducible factor-1αimmunostaining. Calibrated radiochromic flms and GEANT-4 simulations verifed the dosimetry variations inside andoutside the chambers. We irradiated hypoxic human skin fbroblasts (AG01522B) cells with laser-driven protons, conventional protons and reference 225 kVp X-rays to quantify DNA DSB damage and repair under hypoxia. We furthermeasured the oxygen enhancement ratio for cell survival after X-ray exposure in normal fbroblast and radioresistantpatient- derived GBM stem cells.Results: Oxygen measurements showed that our chambers maintained a radiobiological hypoxic environment forat least 45 min and pathological hypoxia for up to 24 h after disconnecting the chambers from the gas supply. Weobserved a signifcant reduction in the 53BP1 foci induced by laser-driven protons, conventional protons and X-rays inthe hypoxic cells compared to normoxic cells at 30 min post-irradiation. Under hypoxic irradiations, the Laser-driven</p>
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