Dosimetry is a vital component of treatment planning in radiation therapy. Methods of radiation dosimetry currently include the use of: ionization chambers, thermoluminescent dosimeters (TLDs), solid-state detectors and radiographic film. However, these methods are inherently either 1D or 2D and their use involves the perturbation of the radiation beam. Although the dose distribution within tissues following radiation therapy treatments can be modeled using computerized treatment planning systems, a need exists for a dosimeter that can accurately measure dose distributions directly and produce 3D dose maps. Some radiation therapy and brachytherapy treatments require mapping the dose distributions in high-resolution (typically < 1 mm). A dosimetry technique that is capable of producing high resolution 3D dose maps of the absorbed dose distribution within tissues is required. Gel dosimetry is inherently a 3D integrating dosimeter that offers high spatial resolution, precision and accuracy. Polymer gel dosimetry is founded on the basis that monomers dissolved in the gel matrix polymerize due to the presence of free radicals produced by the radiolysis of water molecules. The amount of polymerization that occurs within a polymer gel dosimeter can be correlated to the absorbed dose. The gel matrix maintains the spatial integrity of the polymers and hence a dose distribution can be determined by imaging the irradiated polymer gel dosimeter using an imaging modality such as MRI, x-ray computed tomography (CT), ultrasound, optical CT or vibrational spectroscopy. Polymer gel dosimeters, however, suffer from oxygen contamination. Oxygen inhibits the polymerization reaction and hence polymer gel dosimeters must be manufactured, irradiated and scanned in hypoxic environments. Normoxic polymer gel dosimeters incorporate an anti-oxidant into the formulation that binds the oxygen present in the gel and allows the dosimeter to be made under normal atmospheric conditions. The first part of this study was to provide a comprehensive investigation into various formulations of polymer and normoxic polymer gel dosimeters. Several parameters were used to characterize and assess the performance of each formulation of polymer gel dosimeter including: spatial resolution and stability, temporal stability of the R2-dose response, optimal R2-dose response for changes in concentration of constituents and the effects of oxygen infiltration. This work enabled optimal formulations to be determined that would provide greater dose sensitivity. Further work was done to investigate the chemical kinetics that take place within normoxic polymer gel dosimeters from manufacture to post-irradiation. This study explored the functions that each of the constituent chemicals plays in a polymer gel dosimeter. Although normoxic polymer gel dosimeters exhibit very similar characteristics to polyacrylamide polymer gel dosimeters, one important difference between them was found to be a decrease in R2-dose sensitivity over time in the normoxic polymer gel dosimeter compared to an increase in the polyacrylamide polymer gel dosimeters. From an investigation into the function of anti-oxidants in normoxic polymer gel dosimeters, alternatives were proposed. Several alternative anti-oxidants were explored in this study that found that whilst some were reasonably effective, tetrakis (hydroxymethyl) phosphonium chloride (THPC) had the highest reaction rate. THPC was found not only to be an aggressive scavenger of oxygen, but also to increase the dose sensitivity of the gel. Hence, a formulation of normoxic polymer gel dosimeter was proposed, called MAGAT, that comprised: methacrylic acid, gelatin, hydroquinone and THPC. This formulation was examined in a similar fashion to the studies of the other formulations of polymer and normoxic polymer gel dosiemeters. The gel was found to exhibit spatial and temporal stability and an optimal formulation was proposed based on the R2-dose response. Applications such as IVBT require high-resolution dosimetry. Combined with high-resolution MRI, polymer gel dosimetry has potential as a high-resolution 3D integrated dosimeter. Thus, the second component of this study was to commission a micro-imaging MR spectrometer for use with normoxic polymer gel dosimeters and investigate artifacts related to imaging in high-resolutions. Using high-resolution MRI requires high gradient strengths that, combined with the Brownian motion of water molecules, was found to produce an attenuation of the MR signal and hence lead to a variation in the measured R2. The variation in measured R2 was found to be dependent on both the timing and amplitude of pulses in the pulse sequence used during scanning. Software was designed and coded that could accurately determine the amount of variation in measured R2 based on the pulse sequence applied to a phantom. Using this software, it is possible to correct for differences between scans using different imaging parameters or pulse sequences. A normoxic polymer gel dosimeter was irradiated using typical brachytherapy delivery and the resulting dose distributions compared with dose points predicted by the computerized treatment planning system.The R2-dose response was determined and used to convert the R2 maps of the phantoms to dose maps. The phantoms and calibration vials were imaged with an in-plane resolution of 0.1055 mm/pixel and a slice thickness of 2 mm. With such a relatively large slice thickness compared to the in-plane resolution, partial volume effects were significant, especially in the region immediately adjacent the source where high dose gradients typically exist. Estimates of the partial volume effects at various distances within the phantom were determined using a mathematical model based on dose points from the treatment planning system. The normalized and adjusted dose profiles showed very good agreement with the dose points predicted by the treatment planning system.

A systems biology approach was adopted in order to assess various aspects of the disease oral squamous cell carcinoma. Three main aims were addressed; assess the ability of CoCl2 to mimic the hypoxic response in a eukaryotic cell line, assess the role of PDE4D in oral squamous cell carcinoma (OSCC) and the construction of a normoxic/hypoxic mathematical model to identify therapeutic targets.Cancer cells often acquire a revised metabolism which aids in initiation, survival and progression of the tumour. This is predominantly due to the transcription factor HIF-1 which is activated under hypoxic conditions. Certain compounds such as cobalt chloride (CoCl2) have been used extensively to inhibit the degradation of HIF-1α and simulate hypoxia. CoCl2 is likely to have off-target effects on metabolism; these effects were examined when exposing human telomerase reverse transcriptase (hTERT) cells to 100μM CoCl2. Gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) based metabolomics were utilised in combination with ELISA assays for HIF-1α and ATP. Central metabolism was accurately mimicked when hTERT cells were subjected to 100μM CoCl2, however; it was apparent that this concentration of CoCl2 does not induce an equal extent of hypoxia as 1% oxygen. A number of off-target effects of CoCl2 were observed in secondary metabolism, specifically in lipids and fatty acids. In conclusion, CoCl2 should be used with caution as a hypoxic mimicker with the caveat that interpretation of results should be restricted to its effects on central metabolism.The transcription factor CREB has the ability to regulate approximately 4000 genes, a number of which are associated with cancer initiation and progression. Cyclic adenosine monophosphate (cAMP) is required to activate CREB and is partially regulated through its degradation via the enzyme phosphodiesterase type 4D (PDE4D). A homozygous deletion of PDE4D has been associated with OSCC; however; the exact consequence of this deletion has not been fully elucidated. PDE4D was knocked down in the OSCC cell line BicR16 and cellular proliferation, migration, resistance to ionising radiation and central metabolism was investigated using MTT, scratch, clonogenic and GC-MS, respectively. The knockdown resulted in an increase in proliferation, migration and radiation resistance suggesting the role of PDE4D as a TSG. Amino acids, cholesterol, fatty acids, carbohydrates and TCA intermediates were found to be altered in concentration.A mathematical model of glycolysis, TCA and glutaminolysis under normoxia and hypoxia was constructed through the amalgamation of two established models from the literature. New reactions, parameters and metabolite concentrations were added and unnecessary entities were deleted. COmplex PAthway SImulator (COPASI) was utilised to construct the model before validating the model using experimental data from the literature and steady state and flux analyses. Sensitivity analysis and a reduction in external glucose and glutamine were mimicked and the alterations in hypoxic and normoxic metabolism analysed. The reactions vCSII, vGS, vPGK and vGII were identified as potential therapeutic targets which may affect metabolism in hypoxia only. However, certain validation methods proved unsuccessful and hence the model requires further work before attempting the analyses again.

The overall objective of this study was to develop and characterise new normoxic polymer gel formulations for evaluation of complex 3-D treatment volumes for application in radiotherapy dosimetry. Throughout this thesis, the essential characteristics of normoxic polymer gels have been extensively investigated. Studies were performed on the chemical components of the MAGIC gel and an improved formulation was proposed. Various anti-oxidants were studied and different versions of the MAGIC gel with fewer chemicals were developed and named MAGAS and MAGAT gel dosimeters. The ascorbic acid anti-oxidant was found to have a slow oxygen scavenging rate and therefore a delay period between manufacture and irradiation of the MAGAS gel was necessary before the gel became radiation sensitive. Vacuum pumping on the MAGAS gel solution to remove dissolved oxygen was shown to initially increase the R2-dose response and sensitivity of the dosimeter, reducing the time between manufacture and irradiation. Studies of the MAGAS gel for measurement of depth dose showed that MAGAS gel has potential as a clinical radiotherapy dosimetry tool. The radiological properties of MAGIC, MAGAS and MAGAT gels were investigated. Due to their high gelatine and monomer concentration, differences with water were observed for the cross-section ratios for attenuation, energy absorption and collision stopping power coefficient ratios through the therapeutic energy range. It was determined that when using and developing normoxic polymer gels the most important consideration for radiological water equivalence are the mass and relative electron densities. A preliminary study was performed with the hypoxic PAG gel dosimeter combined with tetrakis (hydroxymethyl) phosphonium chloride anti-oxidant to form a normoxic PAG gel dosimeter named PAGAT gel. It was found PAGAT gel compared favourably with previous studies of the hypoxic PAG gel. An extensive study was subsequently undertaken in which PAGAT gel was investigated for a number of essential characteristics. The PAGAT gel formulation showed potential as a normoxic polymer gel for clinical radiotherapy dosimetry, which has a significantly reduced manufacturing time and procedure compared with the hypoxic PAG gel dosimeter. The radiological attenuation properties of the PAGAT and MAGAT gels were investigated as a feasibility study for using x-ray computerised tomography (CT) as an evaluation technique of normoxic polymer gels. CT was shown to have potential as an evaluation tool for measuring the dose response of normoxic polymer gel dosimeters. An investigation was performed on the CT diagnostic dose response of normoxic polymer gels. Normoxic polymer gels were found to have potential for use as a specialised tool in measuring computerised tomography dose index (CTDI) for acceptance testing and quality assurance of CT scanners in diagnostic radiology. These findings provide a significant contribution toward the development and successful implementation of normoxic polymer gel dosimetry to clinical radiotherapy.