Effect of glass compression plate on EBT-XD film dosimetry for pretreatment quality assurance of stereotactic body radiotherapy.

Background: EBT-XD film specially designed for high dose verifications such as stereotactic treatments. The dose response of the film can be affected by several factors, the curly nature of the film being one of them. In this study this curly nature of the film was investigated for stereotactic body radiotherapy (SBRT) plan verifications. Materials and methods: For this study, 18 SBRT (11 prostate, 3 spines, and 4 lungs) cases were enrolled. For all the cases, VMAT plans were created in the Monaco treatment planning system and plan was delivered in Elekta Versa HD linear accelerator and delivered fluence was captured by EBT-XD films. All films were scanned with and without a compression plate. All the films were analyzed using the single-channel film method using the red channel. Results: A significant difference in the gamma passing rates (GPR) for the films scanned with and without the compression plate was observed. The maximum percentage differences in GPR between using and not using a compression plate were 12.7% for 1% 1 mm, 8.1% for 2% 2 mm, 7.5% for 3% 2 mm, and 5% for 3% 3mm criteria. Similarly, the mean %difference in GPR was 5.8% for 1% 1 mm, 2.4% for 2% 2 mm, 1.6% for 3% 2 mm and 0.96% for 3% 3 mm criteria. Conclusion: The results suggest that placing a compression plate over the film during scanning provided a great advantage in achieving a more accurate gamma passing rate irrespective of gamma criteria.

Switching in the expression pattern of actin isoforms marks the onset of contractility and distinct mechanodynamic behavior during cardiomyocyte differentiation.

Differentiation of cardiac progenitor cells (CPC) into cardiomyocytes is a fundamental step in cardiogenesis, which is marked by changes in gene expression responsible for remodeling of the cytoskeleton and in altering the mechanical properties of cells. Here we have induced the differentiation of CPC derived from human pluripotent stem cells into immature cardiomyocytes (iCM) which we compare with more differentiated cardiomyocytes (mCM). Using atomic force microscopy and real-time deformability cytometry, we describe the mechanodynamic changes that occur during the differentiation process and link our findings to protein expression data of cytoskeletal proteins. Increased levels of cardiac-specific markers as well as evolution of cytoskeletal morphology and contractility parameters correlated with the expected extent of cell differentiation that was accompanied by hypertrophic growth of cells. These changes were associated with switching in the balance of the different actin isoforms where ß-actin is predominantly found in CPC, smooth muscle α-actin is dominant in iCM cells and sarcomeric α-actin is found in significantly higher levels in mCM. We link these cytoskeletal changes to differences in mechano-dynamic behavior of cells that translate to changes in Young's modulus that depend on the cell adherence. Our results demonstrate that the intracellular balance of actin isoform expression can be used as a sensitive ruler to determine the stage of differentiation during early phases of cardiomyocyte differentiation that correlates with an increased expression of sarcomeric proteins and is accompanied by changes in cellular elasticity.

Cardiomyocyte mechanodynamics under conditions of actin remodelling.

The mechanical performance of cardiomyocytes (CMs) is an important indicator of their maturation state and of primary importance for the development of therapies based on cardiac stem cells. As the mechanical analysis of adherent cells at high-throughput remains challenging, we explore the applicability of real-time deformability cytometry (RT-DC) to probe cardiomyocytes in suspension. RT-DC is a microfluidic technology allowing for real-time mechanical analysis of thousands of cells with a throughput exceeding 1000 cells per second. For CMs derived from human-induced pluripotent stem cells, we determined a Young's modulus of 1.25 ± 0.08 kPa which is in close range to previous reports. Upon challenging the cytoskeleton with cytochalasin D (CytoD) to induce filamentous actin depolymerization, we distinguish three different regimes in cellular elasticity. Transitions are observed below 10 nM and above 103 nM and are characterized by a decrease in Young's modulus. These regimes can be linked to cytoskeletal and sarcomeric actin contributions by CM contractility measurements at varying CytoD concentrations, where we observe a significant reduction in pulse duration only above 103 nM while no change is found for compound exposure at lower concentrations. Comparing our results to mechanical cell measurements using atomic force microscopy, we demonstrate for the first time to our knowledge, the feasibility of using a microfluidic technique to measure mechanical properties of large samples of adherent cells while linking our results to the composition of the cytoskeletal network. This article is part of a discussion meeting issue 'Single cell ecology'.