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clinically due to the number of variables that need to be investigated to control and optimize the effect. Various groups have investigated different aspects, such as varying NP size or radiation beam energy. Even with knowledge from these studies, there is still a considerable amount of variability in reported findings. Differences are caused by the diversity in cell lines, NPs with their respective coatings, incubated NP concentrations, incubation times, irradiation parameters, as well as the assays used to demonstrate the effects. This has led to variations in the results, where significant enhancements of a factor of 25 were shown by Rahman et al., with Aurovist 1.9 nm gold nanoparticles (AuNPs) at a concentration of 1 mm with bovine aortic endothelial cells (BAEC) and 80 kV X-rays, [3] compared to smaller enhancements shown by Chithrani et al., where they synthesized 50 nm AuNPs at a concentration of approximately 1 nm in HeLa cells and found an enhancement factor of 1.17 with 6 MV X-rays. [4] As well as this, there are also differences in protocols between research groups, in both maintenance of cells and assays reported.A review by Her et al. reported on the different mechanisms associated with NP-enhanced radiotherapy, where the overall effect is a combination of physical, chemical, and biological -7 and U87) and three commercially available nanoparticles (gold, gadolinium, and iron oxide) irradiated by 6 MV X-rays. To assess cell survival, clonogenic assays are carried out for all variables considered, with a concentration of 0.5 mg mL -1 for each nanoparticle material used. This study demonstrates differences in cell survival between nanoparticles and cell line. U87 shows the greatest enhancement with gadolinium nanoparticles (2.02 ± 0.36), whereas MCF-7 cells have higher enhancement with gold nanoparticles (1.74 ± 0.08). Materials with a high atomic number (Z) are shown to cause an increase in the level of cell kill by ionizing radiation when introduced into tumor cells. This study uses in vitro experiments to investigate the differences in radiosensitization between two cell lines (MCFMass spectrometry, however, shows highest elemental uptake with iron oxide and U87 cells with 4.95 ± 0.82 pg of iron oxide per cell. A complex relationship between cellular elemental uptake is demonstrated, highlighting an inverse correlation with the enhancement, but a positive relation with DNA damage when comparing the same nanoparticle between the two cell lines.The ORCID identification number(s) for the author(s) of this article can be found under https://doi.

Despite well-established dosimetry in clinical radiotherapy, dose measurements in pre-clinical and radiobiology studies are frequently inadequate, thus undermining the reliability and reproducibility of published findings. The lack of suitable dosimetry protocols, coupled with the increasing complexity of pre-clinical irradiation platforms, undermines confidence in preclinical studies and represents a serious obstacle in the translation to clinical practice. To accurately measure output of a pre-clinical radiotherapy unit, appropriate Codes of Practice (CoP) for medium energy x-rays needs to be employed. However, determination of absorbed dose to water (Dw) relies on application of backscatter factor (Bw) employing in-air method or carrying out in-phantom measurement at the reference depth of 2 cm in a full backscatter (i.e. 30 × 30 × 30 cm3) condition. Both of these methods require thickness of at least 30 cm of underlying material, which are never fulfilled in typical pre-clinical irradiations. This work is focused on evaluation the effects of the lack of recommended reference conditions in dosimetry measurements for pre-clinical settings and is aimed at extending the recommendations of the current CoP to practical experimental conditions and highlighting the potential impact of the lack of correct backscatter considerations on radiobiological studies.