Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel

Abstract Synchrotron microbeam radiotherapy (MRT) is an innovative cancer treatment that uses micron-sized of ultra-high dose rate spatially fractionated X-rays to effectively control cancer growth while reducing the damage to surrounding healthy tissue. However, the current pre-clinical experiments...

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Main Authors: John Paul O. Bustillo, Elette E.M. Engels, Vincent de Rover, Kiarn Roughley, Julia Rebecca D. Posadas, Elrick T. Inocencio, Danielle Warren, Gordon G. Wallace, Moeava Tehei, Anatoly B. Rosenfeld, Michael L.F. Lerch
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Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-88793-9
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author John Paul O. Bustillo
Elette E.M. Engels
Vincent de Rover
Kiarn Roughley
Julia Rebecca D. Posadas
Elrick T. Inocencio
Danielle Warren
Gordon G. Wallace
Moeava Tehei
Anatoly B. Rosenfeld
Michael L.F. Lerch
author_facet John Paul O. Bustillo
Elette E.M. Engels
Vincent de Rover
Kiarn Roughley
Julia Rebecca D. Posadas
Elrick T. Inocencio
Danielle Warren
Gordon G. Wallace
Moeava Tehei
Anatoly B. Rosenfeld
Michael L.F. Lerch
author_sort John Paul O. Bustillo
collection DOAJ
description Abstract Synchrotron microbeam radiotherapy (MRT) is an innovative cancer treatment that uses micron-sized of ultra-high dose rate spatially fractionated X-rays to effectively control cancer growth while reducing the damage to surrounding healthy tissue. However, the current pre-clinical experiments are commonly limited with the use of conventional two-dimensional cell cultures which cannot accurately model in vivo tissue environment. This study aims to propose a three-dimensional (3D) bioprinting gelatin methacryloyl (GelMA) hydrogel protocol and to characterize 3D bioprinted glioma relative to cell monolayer and spheroid models for experimental MRT using 9L rat gliosarcoma and U87 human glioma. Synchrotron broad-beam (SBB) and MRT beams were delivered to all cell models using 5, 10, and 20 Gy. 3D bioprinting enables the creation of 3D cell models that mimic in vivo conditions using bioinks, biomaterials, and cells. Synchrotron dosimetry, Monte Carlo simulation, in vitro cell viability, and fluorescence microscopy were performed to understand the relationship of the radiation dosimetry with the radiobiological response of different cancer models. Encapsulated gliomas were placed inside 3D printed human and rat phantoms to mimic scattering conditions. Results showed that MRT kills more gliomas relative to SBB for all cell models. The 3D bioprinted culture detected the spatial clustering of dead cells due to MRT high peak doses as seen in fluorescence imaging. The result of this study progresses MRT research by integrating 3D bioprinting techniques in radiobiological experiments. The study’s bioprinting protocol and results will help in reducing the use of animal experiments and possibly in clinical translation of MRT.
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spelling doaj-art-2167274d36a7409c866a5ee24b40bb7a2025-08-20T03:14:03ZengNature PortfolioScientific Reports2045-23222025-04-0115112010.1038/s41598-025-88793-9Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogelJohn Paul O. Bustillo0Elette E.M. Engels1Vincent de Rover2Kiarn Roughley3Julia Rebecca D. Posadas4Elrick T. Inocencio5Danielle Warren6Gordon G. Wallace7Moeava Tehei8Anatoly B. Rosenfeld9Michael L.F. Lerch10Centre for Medical Radiation Physics, University of Wollongong AustraliaCentre for Medical Radiation Physics, University of Wollongong AustraliaCentre for Medical Radiation Physics, University of Wollongong AustraliaCentre for Medical Radiation Physics, University of Wollongong AustraliaDepartment of Physical Sciences and Mathematics, College of Arts and Sciences, University of the Philippines ManilaDepartment of Physical Sciences and Mathematics, College of Arts and Sciences, University of the Philippines ManilaAIIM Facility, Intelligent Polymer Research Institute, ARC Centre of Excellence for Electromaterials Science, University of WollongongAIIM Facility, Intelligent Polymer Research Institute, ARC Centre of Excellence for Electromaterials Science, University of WollongongCentre for Medical Radiation Physics, University of Wollongong AustraliaCentre for Medical Radiation Physics, University of Wollongong AustraliaCentre for Medical Radiation Physics, University of Wollongong AustraliaAbstract Synchrotron microbeam radiotherapy (MRT) is an innovative cancer treatment that uses micron-sized of ultra-high dose rate spatially fractionated X-rays to effectively control cancer growth while reducing the damage to surrounding healthy tissue. However, the current pre-clinical experiments are commonly limited with the use of conventional two-dimensional cell cultures which cannot accurately model in vivo tissue environment. This study aims to propose a three-dimensional (3D) bioprinting gelatin methacryloyl (GelMA) hydrogel protocol and to characterize 3D bioprinted glioma relative to cell monolayer and spheroid models for experimental MRT using 9L rat gliosarcoma and U87 human glioma. Synchrotron broad-beam (SBB) and MRT beams were delivered to all cell models using 5, 10, and 20 Gy. 3D bioprinting enables the creation of 3D cell models that mimic in vivo conditions using bioinks, biomaterials, and cells. Synchrotron dosimetry, Monte Carlo simulation, in vitro cell viability, and fluorescence microscopy were performed to understand the relationship of the radiation dosimetry with the radiobiological response of different cancer models. Encapsulated gliomas were placed inside 3D printed human and rat phantoms to mimic scattering conditions. Results showed that MRT kills more gliomas relative to SBB for all cell models. The 3D bioprinted culture detected the spatial clustering of dead cells due to MRT high peak doses as seen in fluorescence imaging. The result of this study progresses MRT research by integrating 3D bioprinting techniques in radiobiological experiments. The study’s bioprinting protocol and results will help in reducing the use of animal experiments and possibly in clinical translation of MRT.https://doi.org/10.1038/s41598-025-88793-9Synchrotron RadiationMicrobeam Radiation TherapySpatial fractionationBioprintingGliomaGelMA
spellingShingle John Paul O. Bustillo
Elette E.M. Engels
Vincent de Rover
Kiarn Roughley
Julia Rebecca D. Posadas
Elrick T. Inocencio
Danielle Warren
Gordon G. Wallace
Moeava Tehei
Anatoly B. Rosenfeld
Michael L.F. Lerch
Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
Scientific Reports
Synchrotron Radiation
Microbeam Radiation Therapy
Spatial fractionation
Bioprinting
Glioma
GelMA
title Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
title_full Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
title_fullStr Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
title_full_unstemmed Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
title_short Three-dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
title_sort three dimensional bioprinted in vitro glioma tumor constructs for synchrotron microbeam radiotherapy dosimetry and biological study using gelatin methacryloyl hydrogel
topic Synchrotron Radiation
Microbeam Radiation Therapy
Spatial fractionation
Bioprinting
Glioma
GelMA
url https://doi.org/10.1038/s41598-025-88793-9
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