Distinguishing Tendon and Ligament Fibroblasts Based on ¹H Nuclear Magnetic Resonance Spectroscopy

Tendon and ligament (T/L) have been known to be obviously different from each other in tissue level. However, due to the overlapping gene markers, distinction in cellular level has not been clearly verified yet. Recently, the use of nuclear magnetic resonance (NMR) spectroscopy has shown the potential to detect biological markers in cellular level. Therefore, in this study we applied a non-invasive technique based on NMR spectroscopy to establish biomarkers to distinguish between T/L fibroblasts. In addition the cellular morphologies and gene expression patterns were also investigated for comparison through optical microscopy and real-time polymerase chain reaction (PCR). No difference was observed from morphology and real-time PCR results, either as expected. However, we found clear differences in their metabolomic spectra using ¹H NMR spectroscopy. The calculated integral values of fatty acids (with chemical shifts at ~0.9, 1.26, 1.59, 2.05, 2.25, and 2.81 ppm), lactate (~1.33 ppm), and leucine (~2.72 ppm) were significantly different between the two types of fibroblasts. To be specific tendon group exhibited higher level of the metabolite than ligament group. In conclusion, in-cell metabolomic evaluation by NMR technique used in this study is believed to provide a promising tool in distinguishing cell types, especially T/L cells, which cannot be classified by conventional biological assays.

Time Resolution Improvement of MRI Temperature Monitoring Using Keyhole Method

PURPOSE: This study proposes the keyhole method in order to improve the time resolution of the proton resonance frequency(PRF) MR temperature monitoring technique. The values of Root Mean Square (RMS) error of measured temperature value and Signal-to-Noise Ratio(SNR) obtained from the keyhole and full phase encoded temperature images were compared. MATERIALS AND METHODS: The PRF method combined with GRE sequence was used to get MR temperature images using a clinical 1.5T MR scanner. It was conducted on the tissue-mimic 2% agarose gel phantom and swine's hock tissue. A MR compatible coaxial slot antenna driven by microwave power generator at 2.45GHz was used to heat the object in the magnetic bore for 5 minutes followed by a sequential acquisition of MR raw data during 10 minutes of cooling period. The acquired raw data were transferred to PC after then the keyhole images were reconstructed by taking the central part of K-space data with 128, 64, 32 and 16 phase encoding lines while the remaining peripheral parts were taken from the 1st reference raw data. The RMS errors were compared with the 256 full encoded self-reference temperature image while the SNR values were compared with the zero filling images. RESULTS: As phase encoding number at the center part on the keyhole temperature images decreased to 128, 64, 32 and 16, the RMS errors of the measured temperature increased to 0.538, 0.712, 0.768 and 0.845degrees C, meanwhile SNR values were maintained as the phase encoding number of keyhole part is reduced. CONCLUSION: This study shows that the keyhole technique is successfully applied to temperature monitoring procedure to increases the temporal resolution by standardizing the matrix size, thus maintained the SNR values. In future, it is expected to implement the MR real time thermal imaging using keyhole method which is able to reduce the scan time with minimal thermal variations.

Preliminary Study on the MR Temperature Mapping using Center Array-Sequencing Phase Unwrapping Algorithm

PURPOSE: To investigate the feasibility and accuracy of Proton Resonance Frequency (PRF) shift based magnetic resonance (MR) temperature mapping utilizing the selfdeveloped center array-sequencing phase unwrapping (PU) method for non-invasive temperature monitoring. MATERIALS AND METHODS: The computer simulation was done on the PU algorithm for performance evaluation before further application to MR thermometry. The MR experiments were conducted in two approaches namely PU experiment, and temperature mapping experiment based on the PU technique with all the image postprocessing implemented in MATLAB. A 1.5T MR scanner employing a knee coil with T2* GRE (Gradient Recalled Echo) pulse sequence were used throughout the experiments. Various subjects such as water phantom, orange, and agarose gel phantom were used for the assessment of the self-developed PU algorithm. The MR temperature mapping experiment was initially attempted on the agarose gel phantom only with the application of a custom-made thermoregulating water pump as the heating source. Heat was generated to the phantom via hot water circulation whilst temperature variation was observed with T-type thermocouple. The PU program was implemented on the reconstructed wrapped phase images prior to map the temperature distribution of subjects. As the temperature change is directly proportional to the phase difference map, the absolute temperature could be estimated from the summation of the computed temperature difference with the measured ambient temperature of subjects. RESULTS: The PU technique successfully recovered and removed the phase wrapping artifacts on MR phase images with various subjects by producing a smooth and continuous phase map thus producing a more reliable temperature map. CONCLUSION: This work presented a rapid, and robust self-developed center arraysequencing PU algorithm feasible for the application of MR temperature mapping according to the PRF phase shift property.