Advances in radiotherapy, particularly the exploration of alternative radiation types such as carbon ions have updated our understanding of its effects and applicability on chondrosarcoma cells. Here we compare the optical effects produced by carbon ions (CI) and X-rays (XR) radiations on chondrosarcoma cells nuclei and set an automated method for evaluating the radiation-induced alterations without the need of chemical marking. Hyperspectral images (HSI) of SW1353 chondrosarcoma line carry detectable optical changes of the cells irradiated either with CI or XR compared to non-irradiated ones (REF). The differences between the spectral profiles of CI, XR and REF nuclei classes led to partitioning the HSIs into spectral sub-images. The changes are detected by support vector machine (SVM) classifiers whose performances are evaluated by the most used point metrics: sensitivity (SEN), accuracy (ACC), and precision (PREC), applied on spatial feature values. Specific interaction mechanisms by radiation type reveal distinct subintervals where HSIs changes are more prominent, and the classifiers perform at best. For CI the best classifiers are obtained for sub-images in the interval (424-436 nm), while for XR the best classifiers are obtained for sub-images in the interval (436-445 nm). The classifiers work better with texture features than roughness features in both cases. The classifier with the best SEN point metric in the testing phase is the most suitable to measure the irradiation efficiency irrespective of the radiation type. The altered nuclei are easier to discriminate when irradiated with CI than with XR. The study proves that SVM with optical data offers a rapid, automated, and label-free method for evaluating radiation-induced alterations in chondrosarcoma nuclei, thereby enabling effective analysis of extensive data.

To overcome chondrosarcoma’s (CHS) high chemo- and radioresistance, we used polyethylene glycol-encapsulated iron oxide nanoparticles (IONPs) for the controlled delivery of the chemotherapeutic doxorubicin (IONP_DOX) to amplify the cytotoxicity of proton radiation therapy. Human 2D CHS SW1353 cells were treated with protons (linear energy transfer (LET): 1.6 and 12.6 keV/µm) with and without IONP_DOX. Cell survival was assayed using a clonogenic test, and genotoxicity was tested through the formation of micronuclei (MN) and γH2AX foci, respectively. Morphology together with spectral fingerprints of nuclei were measured using enhanced dark-field microscopy (EDFM) assembled with a hyperspectral imaging (HI) module and an axial scanning fluorescence module, as well as scanning electron microscopy (SEM) coupled with energy-dispersive X-Ray spectroscopy (EDX). Cell survival was also determined in 3D SW3153 spheroids following treatment with low-LET protons with/without the IONP_DOX compound. IONP_DOX increased radiosensitivity following proton irradiation at both LETs in correlation with DNA damage expressed as MN or γH2AX. The IONP_DOX–low-LET proton combination caused a more lethal effect compared to IONP_DOX–high-LET protons. CHS cell biological alterations were reflected by the modifications in the hyperspectral images and spectral profiles, emphasizing new possible spectroscopic markers of cancer therapy effects. Our findings show that the proposed treatment combination has the potential to improve the management of CHS.