Accurate, robust and harmonized implementation of morpho-functional imaging in treatment planning for personalized radiotherapy.

In this work we present a methodology able to use harmonized PET/CT imaging in dose painting by number (DPBN) approach by means of a robust and accurate treatment planning system. Image processing and treatment planning were performed by using a Matlab-based platform, called CARMEN, in which a full Monte Carlo simulation is included. Linear programming formulation was developed for a voxel-by-voxel robust optimization and a specific direct aperture optimization was designed for an efficient adaptive radiotherapy implementation. DPBN approach with our methodology was tested to reduce the uncertainties associated with both, the absolute value and the relative value of the information in the functional image. For the same H&N case, a single robust treatment was planned for dose prescription maps corresponding to standardized uptake value distributions from two different image reconstruction protocols: One to fulfill EARL accreditation for harmonization of [18F]FDG PET/CT image, and the other one to use the highest available spatial resolution. Also, a robust treatment was planned to fulfill dose prescription maps corresponding to both approaches, the dose painting by contour based on volumes and our voxel-by-voxel DPBN. Adaptive planning was also carried out to check the suitability of our proposal. Different plans showed robustness to cover a range of scenarios for implementation of harmonizing strategies by using the highest available resolution. Also, robustness associated to discretization level of dose prescription according to the use of contours or numbers was achieved. All plans showed excellent quality index histogram and quality factors below 2%. Efficient solution for adaptive radiotherapy based directly on changes in functional image was obtained. We proved that by using voxel-by-voxel DPBN approach it is possible to overcome typical drawbacks linked to PET/CT images, providing to the clinical specialist confidence enough for routinely implementation of functional imaging for personalized radiotherapy.

Use of X-Ray Computed Tomography for Ice Detection Applied to Organ Cryopreservation.

One of the main problems in the cryopreservation of biological samples is the formation of ice and the consequent mechanical damage to cells and tissues, due to the crystalline structure of ice and its associated mechanical damage. It is necessary to detect this deleterious formation of ice, especially in tissues and organs, because of their large volume and the complexity of their vascular system in the case of bulky organs. In this work, we propose the use of X-ray Computed Tomography (CT) to detect this ice formation inside tissues and organs. To achieve this aim, rabbit kidneys were loaded with cryoprotectant solutions containing Me2SO at low temperatures (below -140°C). Drops of water with a volume between 2 and 8 µL were then introduced inside the organs. Finally, the rabbit kidneys were cooled to -196°C. Volumes of ice of up to 1 µL were detected in our CT device, with a resolution of up to 50 µm, validating the proposed technology. On the contrary, we analyzed bovine ovarian tissues cryopreserved with a controlled-rate slow-cooling protocol. CT images showed the different structure on the extracellular ice formation according to the procedure, and even the intracellular ice that can be formed in the tissues. These positive results have a straightforward application in the control of the formation of ice, of significant importance for the creation of biobanks.

An optimized controlled rate slow cooling protocol for bovine ovarian tissue cryopreservation by means of X-ray computed tomography.

Cryopreservation and subsequent transplantation of ovarian tissue is the only option to preserve fertility in certain patients facing gonadotoxic treatment. So far, cryopreservation of ovarian tissue has been carried out mostly by a controlled rate slow cooling process, typically known as slow freezing. Even though there are still some concerns about the iatrogenic damage on the follicle population, this technique has been used in the more than 100 live births reported to date. It is well known that the control of the cryoprotectant loading in the tissue is crucial to in a cryopreservation procedure. We have used the technology of X-ray computed tomography to assess the concentration and distribution of dimethyl sulfoxide (one of the cryoprotectants most used in fertility preservation) inside pieces of bovine ovarian tissue after its cryopreservation. The low voltage used in our device (75 kV) and the high electronic density of this cryoprotectant makes the X-ray attenuation proportional to its concentration. By assessing and comparing the permeation and homogeneity of the cryoprotectant inside ovarian tissue fragments subjected to a controlled rate slow cooling process, we have characterized the effect of variations in the main parameters involved in the process, with the goal of achieving an optimized protocol with higher permeation of the cryoprotectant in the tissue. The most promissory results were obtained by increasing the initial concentration of dimethyl sulfoxide in the vehicle solution from 10 to 20%v/v.

Structural, optical and X-ray attenuation properties of Tb3+:BaxCe1-xF3-x (x = 0.18-0.48) nanospheres synthesized in polyol medium.

Uniform Ba0.18Ce0.82F2.82 nanospheres have been obtained after aging a solution of barium and cerium nitrates and sodium tetrafluoroborate in a mixture of ethylene glycol and water at 120 °C for 20 hours. The diameter of the spheres could be tailored from 65 nm to 80 nm by varying the NaBF4 concentration while maintaining their colloidal stability in aqueous suspension. Increasing the aging temperature led to a phase transformation from hexagonal to cubic symmetry and to a concomitant increase of the Ba/Ce ratio, which reached a value close to the nominal one (50/50) at 240 °C. The same method was successful in obtaining Tb3+-doped nanospheres with homogeneous cation distribution and the same morphological features as the undoped material. An intense green emission was observed after the excitation of the Tb3+-doped samples through the Ce3+-Tb3+ energy transfer (ET) band. The ET efficiency increased with increasing Tb content, the maximum emission being observed for the 10% Tb-doped nanospheres. Aqueous suspensions of the latter sample showed excellent X-ray attenuation values that were superior to those of an iodine-based clinically approved contrast agent. Their fluorescence and X-ray attenuation properties make this material a potential dual bioprobe for luminescence bioimaging and X-ray computed tomography.

Ovarian tissue cryopreservation by stepped vitrification and monitored by X-ray computed tomography.

Ovarian tissue cryopreservation is, in most cases, the only fertility preservation option available for female patients soon to undergo gonadotoxic treatment. To date, cryopreservation of ovarian tissue has been carried out by both traditional slow freezing method and vitrification, but even with the best techniques, there is still a considerable loss of follicle viability. In this report, we investigated a stepped cryopreservation procedure which combines features of slow cooling and vitrification (hereafter called stepped vitrification). Bovine ovarian tissue was used as a tissue model. Stepwise increments of the Me2SO concentration coupled with stepwise drops-in temperature in a device specifically designed for this purpose and X-ray computed tomography were combined to investigate loading times at each step, by monitoring the attenuation of the radiation proportional to Me2SO permeation. Viability analysis was performed in warmed tissues by immunohistochemistry. Although further viability tests should be conducted after transplantation, preliminary results are very promising. Four protocols were explored. Two of them showed a poor permeation of the vitrification solution (P1 and P2). The other two (P3 and P4), with higher permeation, were studied in deeper detail. Out of these two protocols, P4, with a longer permeation time at -40 °C, showed the same histological integrity after warming as fresh controls.

Rapid and simplified synthesis of [18F]Fluoromisonidazole and its use in PET imaging in an experimental model of subarachnoid hemorrhage.

Cerebral damage secondary to the vasospasm due to subarachnoid hemorrhage (SAH) is an important cause of morbid-mortality. We propose the use of the PET tracer [18F]Fluoromisonidazole to visualize the hypoxia due to the vasospasm. On the other hand [18F]Fluoromisonidazole synthesis process was optimized, avoiding HPLC purification using SPE cartridges instead, and reducing some synthesis steps. [18F]Fluoromisonidazole in vitro stability was tested for ten hours, and in vivo PET/CT images showed higher cerebral uptake in hemorrhagic animals than in control rats.

Assessment of the cryoprotectant concentration inside a bulky organ for cryopreservation using X-ray computed tomography.

Cryoprotection of bulky organs is crucial for their storage and for subsequent transplantation. In this work we demonstrate the capability of the X-ray computed tomography (CT) as a non-invasive method to measure the cryoprotectant (cpa) concentration inside a tissue or an organ, specifically for the case of dymethil sulfoxide (Me2SO). It is remarkable that the use of Me2SO has been leader in techniques of cells and tissues cryopreservation. Although CT technologies are mainly based in density differences, and many cpas are alcohols with densities similar to water, the use of very low energies as acceleration voltage (∼70 kV) and the sulfur atom in the molecule of Me2SO makes possible the visualization of this cpa inside tissues. As result we obtain a CT signal proportional to the Me2SO concentration with a spatial resolution up to 50 µm in the case of our device.