ABSTRACT vii

Abstract

NMR techniques have seen widespread use in the characterization of biological systems.

Due to the different environments experienced by spins, the T1 and T2 relaxation behavior

can differ significantly. Furthermore, the local environment can be altered by the

introduction of paramagnetic agents that alter the bulk magnetic susceptibility of a

sample, causing drastic changes in the relaxation behavior of spins. For spins situated in

restricted spaces, the diffusion characteristics as well as the relaxation properties of these

spins can differ from system to system.

Normally, the relaxation (T1 and T2) and diffusion measurements are performed

independently and system parameters are determined solely on the basis of the

independent behavior of the relaxation or diffusion. In the following experiments, these

parameters (T1, T2, and apparent diffusion coefficient) were used to characterize

biological systems. Furthermore, multi-dimensional analysis was performed on a system

where combined relaxometry and diffusimetry were used to characterize the system as a

function of both parameters.

Radiotherapy studies in mice using perfluoro-15-crown-5-ether showed a decrease in pO2

following a single large dose of radiation. In conjunction with spectroscopic data,

Inversion-Recovery Echo-Planar-Imaging data were collected at 1-3 hours, 10-13 hours,

and 19-26 hours post irradiation, and T1-maps generated in order to display localized
ABSTRACT viii

changes in pO2. The calculated T1-maps were then weighted by their respective M0-maps

to find the weighted average of the T1-maps, and an equivalent pO2 of the tumor was then

calculated from the weighted average. Untreated control animals that were subjected to

the same time course showed no evidence of pO2 decline, while the tumors irradiated

with a single dose of 6 MeV electrons showed a decline in pO2 by approximately 9 torr

almost immediately after irradiation. The calculation of pO2 using the weighted average

of the T1-maps was not only highly correlated to the spectroscopic measurements, it was

approximately equivalent to the spectroscopic measurements. It is speculated that the

decrease in the tissue oxygenation following radiation therapy is due to vascular damage

caused by such a high dose of radiation, or edema within the interstitium of the tumor.

Edema can cause the interstitial pressure to increase, resulting in vascular collapse. This

in turn would lead to decreased perfusion and thus decreased oxygen delivery.

Water diffusion-coefficient mapping was used in conjunction with 19F inversion-recovery

echo-planar imaging (IR-EPI) of a sequestered perfluorocarbon (PFC) emulsion to

investigate the spatial correlation between the diffusion coefficient of water and the tissue

oxygen tension (pO2) in radiation-induced fibrosarcoma (RIF-1) tumors (n = 11). The

diffusion-time-dependent apparent diffusion coefficient, D(t), was determined by

acquiring diffusion coefficient maps at 20 different diffusion times. Maps at four

representative time points in different regions of the D(t) curve were selected for final

analysis. An intravenously administered PFC emulsion, perfluoro-15-crown-5-ether, was

used to generate the pO2 maps. D(t) and pO2 data were acquired with the animal
ABSTRACT ix

breathing either air or carbogen (95% O2 – 5% CO2) to investigate the effects of

increased tumor pO2 on D(t). The average increase in tumor pO2 was 22 torr when the

breathing gas was changed from air to carbogen. Correlating plots generated from pixel

data for D(t)(air breathing) versus D(t)(carbogen breathing) showed little deviation from

a slope of unity. Correlation plots of D(t) versus pO2 indicate that no correlation is

present between these two parameters. This study also confirms that necrotic tissue was

best differentiated from viable tumor tissue based on D(t) maps at long diffusion times.

Diffusion-signal-attenuation curves in yeast-cell suspensions show non-monoexponential

signal decay that is assumed to arise from separate compartmental contributions to the

overall signal. However, restricted diffusion effects also give rise to non-

monoexponential signal decay and are difficult to separate from compartmental signal

contributions. Combined relaxometry and diffusion measurements allows differentiation

between compartmental diffusion constants by first separating the compartmental

contributions on the basis of differences in their respective relaxation times. Diffusion-

weighted inversion-recovery spin-echo experiments were carried out at different b-

values. Intra- and extracellular compartments in yeast-cell suspensions were separated on

the basis of T1 relaxation by adding an MR contrast agent to the extracellular space.

Once the compartmental signals were distinguished on the basis of T1, the relative signal

attenuation for each compartment was used to calculate the separate compartmental

ADCs. With this method, even compartmental diffusion coefficients with similar values

can be distinguished.
ABSTRACT x

Water diffusion measurements were made on rabbit Achilles tendon to determine their

behavior during static tensile loading and unloading. Tendons previously stored frozen in

phosphate-buffered saline (PBS) were sequentially loaded with 0.4, 5, 10, and 0.4 N

loads. The apparent diffusion coefficient (ADC) was measured perpendicular ( ADC⊥ )

and parallel ( ADC || ) to the fiber orientation at diffusion times of 10, 30, and 60 ms for

each load. ADC⊥ and ADC || increased with increasing load for all samples. New

samples of freshly-harvested tendons were loaded with a static 5-N load for five minutes

and then unloaded for 30 minutes to determine the effects of loading and unloading. The

ADC ⊥ of fresh tendon was studied as a function of loading and unloading. The ADC ⊥

increased with load for all samples. This increase is attributed to the extrusion of tendon

water into a bulk phase outside the tendon. The recovery of the tendon upon unloading

exhibited a reversal of the ADC ⊥ back to the baseline value. This recovery is attributed

to the water moving from the bulk phase to the bound phase. The recovery followed a

slower time course than the extrusion of water. It was also found that the phosphate-

buffered saline caused the tendon to swell. This method can be used both to detect

structural changes in tendon under tensile loading and to study the transport of water in

tendon.