Diffusion Tensor Imaging Detects Functional and Structural Alterations in Post-Traumatic Distal Tibial Physeal Bars: A Preliminary Study.

Objective To investigate the value of diffusion tensor imaging (DTI) at high b value for unilateral middle cerebral artery (MCA) occlusive disease in patients without obvious infarct lesions on conventional MR imaging.Methods DTI at high b value (2200 s/mm2 ) was performed using a 3.0 Tesla MR scanner in 34 patients with unilateral middle cerebral artery occlusion,who had no obvious infarct lesions on conventional MR imaging. Fractional anisotropy (FA),apparent diffusion coefficient (ADC),axial diffusivity (eigenvalue λ1) and radial diffusivity (eigenvalues λ2,λ3) were measured at the ipsilateral and contralateral corona radiata,anterior and posterior limbs of the internal capsule,cerebral peduncle and pons in all subjects.Mean ADC,FA,λ1,λ2 and λ3 values of corona radiata,anterior and posterior limbs of the internal capsule,cerebral peduncle and pons were compared between the ipsilateral and contralateral MCA territory by t test. Results Among the 34 patients,left MCA occlusion in M1 segment occurred in 16 patients and right MCA occlusion in Ml segment occurred in 18 patients.At the ipsilateral corona radiata,mean FA,ADC,λ1,λ2 and λ3 were 0.419 ±0.032,(5.975 ±0.272) × 10 3,(5.704 ±0.365) ×10-3,(6.412 ±0.368) × 10-3 and (6.605 ±0.343) × 10-3 mm2/s,respectively.At the contralateral corona radiata,mean FA,ADC,λ1,λ2 and λ3 were 0.443 ± 0.033,(5.804 ± 0.282) × 10 -3,(5.651 ±0.350) × 10-3,(6.099 ±0.353) × 10-3 and(6.372 ±0.355) × 10-3 mm2/s,respectively.At the ipsilateral corona radiata,mean FA was significantly decreased(t =11.614,P ＜0.01),and mean ADC (t=12.421,P＜0.01),λ1(t =7.447,P＜0.01),λ2(t=10.244,P＜0.01) and λ3(t=9.890,P＜0.01) were significantly increased.At the ipsilateral anterior and posterior limb of the internal capsule,mean FA were 0.609 ±0.026 and 0.674 ±0.033,λ1 were(5.330 ±0.462) × 10 -3 and(5.171 ±0.456) ×10-3 mm2/s,respectively.At the contralateral anterior and posterior limb of the internal capsule,FA were 0.622 ±0.026 and 0.694 ±0.034,λ1 were(5.064 ± 0.448) × 10 -3 and(4.924 ± 0.365) × 10 -3 mm2/s,respectively.Mean FA was significantly decreased (t =7.823,8.013,all P ＜ 0.01) and mean λ1 was significantly increased (t =7.811,8.800,all P ＜0.01) at the ipsilateral anterior and posterior limbs of the internal capsule.There was no significant difference in ADC,λ2 and λ3 value between the ipsilateral and contralateral sides.And all the DTI parameters,including mean ADC,FA,λ1,λ2 and λ3 values,showed no statistical difference between both sides of cerebral peduncle and pons.Conclusion DTI at high b valuc can provide useful information for visualizing ischemic white matter injury in patients without obvious infarct lesions on conventional MR imaging. Key words: Infarction, middle cerebral artery; Diffusion magnetic resonance imaging

Diffusion tensor (DT) imaging is an emerging magnetic resonance (MR) imaging technique for evaluating the microstructure of well-organized biologic tissues such as muscles and nerves. DT imaging provides information about tissue microstructure by producing three-dimensional maps of water molecule movements. The two main parameters of measurement at DT imaging, fractional anisotropy and the apparent diffusion coefficient, allow quantitation of architectural changes occurring in tissue. These parameters are modified in the presence of cervical spondylotic myelopathy, cervical spine trauma, carpal tunnel syndrome, lumbar nerve compression, peripheral nerve tumors, and muscle ischemia. Their alteration may be observed at DT imaging even when no abnormality is seen at conventional MR imaging, a fact that suggests that DT imaging allows the detection of abnormalities at an earlier stage of injury. Experimental studies in animals have shown that DT imaging consistently allows identification of pathophysiologic alterations in tissue that correlate with histologic findings. Tractographic images accurately depict both normal and abnormal diffusion in anatomic structures such as the thigh and pelvic muscles, cervical spine, and lumbar nerves. Patients with chronic diseases also may benefit from follow-up evaluation with DT imaging, although DT imaging sequences must be further adapted to improve the evaluation of specific anatomic regions by reducing artifacts, optimizing spatial resolution, and minimizing acquisition time. Given its proven potential for use in identifying abnormalities that are otherwise identifiable only with electrophysiologic and histopathologic studies, and with future technical improvements, DT imaging could soon become a standard method for early diagnosis, management, and follow-up of disease in the spine, muscles, and peripheral nerves.

Conventional breast magnetic resonance imaging (MRI), including dynamic contrast-enhanced MR mammography (DCE-MRM), may lead to ambiguous diagnosis and unnecessary biopsies. To investigate the contribution of proton MR spectroscopy (1H-MRS) combined with diffusion tensor imaging (DTI) metrics in the discrimination between benign and malignant breast lesions. Fifty-one women with known breast abnormalities from conventional imaging were examined on a 3T MR scanner. DTI was performed during breast MRI, and fractional anisotropy (FA) and apparent diffusion coefficient (ADC) were measured in the breast lesions and the contralateral normal breast. FA and ADC were compared between malignant lesions, benign lesions, and normal tissue. 1H-MRS was performed after gadolinium administration and choline peak was qualitatively evaluated. In our study 1H-MRS showed a sensitivity of 93.5%, specificity 80%, and accuracy 88.2%. FA was significantly higher in breast carcinomas compared to benign lesions. However, no significant difference was observed in ADC between benign and malignant lesions. The combination of Cho presence and FA achieved higher levels of accuracy and specificity in discriminating malignant from benign lesions over Cho presence or FA alone. In conclusion, applying DTI and 1H-MRS together, adds incremental diagnostic value in the characterization of breast lesions and may sufficiently improve the low specificity of conventional breast MRI.

To determine the role of diffusion tensor imaging (DTI) metrics as biomarkers for the therapeutic effects of mesenchymal stem cells (MSCs) in acute peripheral nerve injury. Forty-four adult rats received subepineurial microinjection of MSCs (n = 22) or phosphate buffered saline (PBS, n = 22) 1 week after the sciatic nerve trunk crush injury. Sequential fat-suppressed T2-weighted imaging, T2 measurement, DTI and sciatic nerve functional assessment were performed at a 3.0 Tesla MR unit over an 8-week follow-up, with histological assessments performed at regular intervals. The sciatic nerve function index, T2 value, and DTI metrics, including fractional anisotropy (FA), axial diffusivity, radial diffusivity (RD), and mean diffusivity values of the distal stumps of crushed nerves were measured and compared between the two groups. Nerves treated with MSCs showed better functional recovery and exhibited more pronounced nerve regeneration compared with nerves treated with PBS. T2 values in nerves treated with MSCs or PBS showed a similar change pattern (P = 0.174), while FA and RD values in nerves treated with MSCs showed more rapid return (one week earlier) to baseline level than nerves treated with PBS (P = 0.045; 0.035). Nerves treated with MSCs had higher FA and lower RD values than nerves treated with PBS during the period from 2 to 3 weeks after surgery (P ≤ 0.0001, 0.004; P = 0.004, 0.006). FA and RD values derived from DTI might be used as sensitive biomarkers for detecting the therapeutic effect of stem cells in acute peripheral nerve crush injuries. 2 J. Magn. Reson. Imaging 2017;45:855-862.

To compare the influence of two different regions of interest (ROIs) on diffusion tensor metrics in dystrophic thigh muscles using acustom-made (whole muscle) ROI including and aselective ROI excluding areas of fatty replacement. Diffusion tensor imaging (DTI) and chemical-shift-encoded water-fat magnetic resonance imaging (MRI) of the thigh was conducted on a3-Tesla system in 15cases with muscular dystrophy and controls. The ROIs were chosen according to patterns of fatty replacement on co-registered axial DTI and gradient echo sequence (GRE) images. Fractional anisotropy (FA), apparent diffusion coefficient (ADC), fiber track length (FTL), and muscle fat fractions (MFF) were compared between both ROI segmentations. These comparisons, muscle-specific correlation coefficients, and the influence of ROI localization on tensor metrics were derived based on linear mixed effects regression models. In the cases ahigh correlation was observed for ADC and FA with MFF using acustom ROI. The correlation was weaker but still significant with aselective ROI method. Using the custom ROI, FTL correlated significantly with MFF in 3 out of 4muscles (r ≤ -0.51). Acorrelation was not found for the selective ROI method. Interaction analysis revealed that the association of ADC and FA with MFF was not significantly influenced by the ROI localization. For FTL the ROI localization significantly reduced the negative association with MFF. The DTI metrics and FTL of custom ROI segmentation are significantly influenced by MFF. Contrary to ADC and FA, the effect of MFF on FTL is significantly reduced when applying selective ROI segmentation, which could therefore be abetter option for MR tractography.

Introduction We present the first study in literature that documents the changes in diffusion tensor imaging (DTI) indices and tractography after surgical decompression in patients with cervical myelopathy. DTI, an established tool for tracking nerve fibers in brain, is now widely evaluated for its usefulness in spine pathologies. Although there are many studies directly comparing DTI indices of control and cervical myelopathic patients, there are no studies that have evaluated the prognostic applicability of DTI indices. The present study analyzes the pre- and postdecompression DTI indices and tractography in cervical myelopathic patients and its prognostic value. Materials and Methods Total 56 subjects including 16 cervical myelopathy patients who required surgical decompression (mean age: 53.5 ± 12.3 years, males: 15, females: (1) and forty normal controls (mean age: 38.18 ± 13.74 years, males: 20, females: 20) were included in the study. DTI of cervical spine was obtained using an axial single shot echo-planar parallel diffusion-weighted imaging sequence in a 1.5 tesla magnetic resonance imaging (MRI). Apparent diffusion coefficient (ADC), fractional anisotropy (FA), and Eigen values (E1, E2, and E3) were obtained at each region of interest (cervical disk level) from C1-C2 to C7-T1. The DTI indices of myelopathic patients were compared with normal volunteers after matching was done based on sex, cord level, and cord size. Postoperatively, the patients were followed up after 6 months and DTI scan was performed again. The clinical improvement and the DTI indices observed at the follow-up were compared with the preoperative values. Results Analysis showed significant differences between the controls and patients in all the indices (p<0.05) at the site of compression. At the compressed levels, FA decreased while all the other indices increased as compared to the controls. The mean DTI indices of myelopathic patients before surgery at the sites of spinal cord compression were as follows: FA (0.49 ± 0.092), ADC (1.8 ± 0.32 µm2/ms), E1 (2.80 ± 0.44 µm2/ms), E2 (1.63 ± 0.45 µm2/ms), E3 (0.95 ± 0.23 µm2/ms). The average DTI indices of matched controls were as follows: FA (0.55 ± 0.033), ADC (1.47 ± 0.066 µm2/ms), E1 (2.49 ± 0.150 µm2/ms), E2 (1.16 ± 0.050 µm2/ms), E3 (0.77 ± 0.058 µm2/ms. The Nurick grading distribution in patients before surgery was as follows: Grade 2 = 2, Gr 3 = 8, Gr 4 = 2, Gr 5 = 4. Postoperatively, all the patients had improvement of neurology by at least one Nurick grade except for one patient who had no improvement. The Nurick grading distribution in patients during 6 months follow-up after surgery was as follows: Grade 0 = 3, Gr 1 = 6, Gr 2 = 2, Gr 3 = 3, Gr 4 = 2. Mean postoperative values of ADC, E1, and E2 showed improvement towards normal values especially in patients with preoperative Nurick grade 2 and 3 (Table 1). Patients with Nurick grade 4 and 5 did not show any significant change. The mean FA and E3 values worsened when compared to preoperative DTI indices. The postoperative DTI indices for the patients who had neurological improvement are as follows: FA (0.45 ± 0.087), ADC (1.73 ± 0.31 µm2/ms), E1 (2.59 ± 0.28 µm2/ms), E2 (1.56 ± 0.38 µm2/ms), E3 (1.07 ± 0.31 µm2/ms. Tractography also showed interruption of nerve fibre tract continuity at the level of severe compression in patients with myelopathy. At levels of moderate compression over the spinal cord, the changes in direction of nerve fibres due to bulging disc-osteophytes were evident as changes in color coding of the tracts. After surgical decompression, the restitution of fibre tract pathways was clearly demonstrated in DT tractography. Conclusion For the first time, the prognostic applicability of DTI by comparing pre- and postoperative indices of cervical myelopathy patients has been described. All preoperative DTI indices at compressed levels showed significant differences from controls indicating abnormality in neuronal integrity. All but one of the patients showed clinical improvement at 6 months. The postoperative DTI values (ADC, E1 and E2) of patients with Nurick grading 2 and 3 were observed to approach normal values indicating restoration of linear diffusion in neuronal tracts. DTI indices in patients with Nurick grade 4 and 5 remained same or worsened more. This could probably indicate persistent axonal damage. At 6 months postsurgery, DTI indices show evidence of improvement towards normal values, even though the values did not match the true extent of clinical improvement. Further DTI evaluation at 12 months and 2 years is required to analyze the extent of usefulness of DTI indices in evaluating cervical myelopathy. The results suggest that DTI indices, especially ADC and Eigen vectors can be promising indicators in the evaluation and prognostic assessment of myelopathy patients. I confirm having declared any potential conflict of interest for all authors listed on this abstract Yes Disclosure of Interest None declared

Global White Matter Changes and Associations with Cognition in Pediatric Sickle Cell Disease

Our aims were to (1) assess the corticospinal tracts (CSTs) in infants with focal injury and healthy term controls using probabilistic tractography and (2) to correlate the conventional magnetic resonance imaging (MRI) and tractography findings in infants with focal injury with their later motor function. We studied 20 infants with focal lesions and 23 controls using MRI and diffusion tensor imaging. Tract volume, fractional anisotropy (FA), apparent diffusion coefficient (ADC) values, axial diffusivity and radial diffusivity (RD) of the CSTs were determined. Asymmetry indices (AIs) were calculated by comparing ipsilateral to contralateral CSTs. Motor outcome was assessed using a standardized neurological examination. Conventional MRI was able to predict normal motor development (n = 9) or hemiplegia (n = 6). In children who developed a mild motor asymmetry (n = 5), conventional MRI predicted a hemiplegia in two and normal motor development in three infants. The AIs for tract volume, FA, ADC and RD showed a significant difference between controls and infants who developed a hemiplegia, and RD also showed a significant difference in AI between controls and infants who developed a mild asymmetry. Conventional MRI was able to predict subsequent normal motor development or hemiplegia following focal injury in newborn infants. Measures of RD obtained from diffusion tractography may offer additional information for predicting a subsequent asymmetry in motor function.

The aim is to investigate whether there is a microstructural change in the white matter pathways in patients with spontaneous intracranial hypotension (SIH). Additionally, the relationship between conventional magnetic resonance imaging (MRI) findings and diffusion tensor imaging (DTI) parameters is determined. Thirty patients diagnosed with SIH and 31 control patients (between January 2019 and February 2024) were included in the case-control study. MRI findings constituting the Bern score were evaluated in contrast-enhanced brain MRI. Ten different white matter pathways were evaluated with the following DTI parameters: fractional anisotropy (FA), apparent diffusion coefficient (ADC), axial diffusion (AD), and radial diffusion (RD). We also examined the association between MRI findings and DTI parameters. In the cingulum, a decreased FA value (Z = -3.18, p = 0.002) was observed in patients with SIH, indicating disruption of white matter integrity and weakened synaptic transmission in this region. In the corona radiata, significant reductions were observed in both FA (Z = -5.67, p < 0.001) and AD (Z = -3.59, p < 0.001), alongside an increase in RD (Z = -4.99, p < 0.001). The concomitant increase in RD further supports the idea of potential damage to the myelin sheath, which, together with other findings, may reflect disruptions in white matter architecture that underlie the cognitive and motor deficits frequently seen in patients with SIH. In the splenium of the corpus callosum, a significant reduction in AD (Z = -3.00, p = 0.007) was also noted. In the anterior limb of the internal capsule (ALIC), both FA (Z = -4.57, p < 0.001) and AD (Z = -2.73, p = 0.012) values decreased, while RD increased (Z = -2.93, p = 0.007). These alterations suggest the involvement of motor and sensory pathways. In the posterior limb of the internal capsule (PLIC), significant reductions were found in both FA (Z = -5.59, p < 0.001) and AD (Z = -3.82, p < 0.001), along with an increase in RD (Z = -5.49, p < 0.001). In the corticospinal tract, patients with SIH exhibited a decrease in FA (Z = -4.15, p < 0.001), an increase in ADC (Z = -2.87, p = 0.008), and an increase in RD (Z = -3.73, p < 0.001). These findings represent significant microstructural changes reflecting a reduction in the integrity of motor pathways in white matter. In the middle cerebellar peduncle (MCP), patients with SIH showed increased ADC (Z = -4.78, p < 0.001) and RD (Z = -3.23, p = 0.002). Similarly, in optic radiation, there was decreased FA (Z = -4.55, p < 0.001) and AD (Z = -2.59, p = 0.018), with increased RD (Z = -2.95, p = 0.007). In the correlation analysis, positive correlations were found between suprasellar distance and AD values in both the ALIC (rs = 0.451, p = 0.012) and MCP (rs = 0.416, p = 0.022). These findings suggest that structural changes and anatomical shifts, especially in the suprasellar region, may be associated with microstructural changes in the adjacent white matter pathways. We revealed microstructural changes in white matter in SIH. The alterations in DTI parameters may suggest that the white matter microarchitecture is changed as a result of mechanical compression of the axons in the white matter pathways due to brain prolapse and venous congestion.

Application of data harmonization and tract-based spatial statistics reveals white matter structural abnormalities in pediatric patients with focal cortical dysplasia

This thesis aimed to investigate the feasibility of two diffusion imaging techniques, Diffusion Tensor Imaging (DTI) and Diffusion Kurtosis Imaging (DKI), on detecting subtle physiological or pathological microstructural changes in normal-appearing neural tissues of human central nervous system. &#13;\n At first, ten patients with neuromyelitis optica (NMO) and twelve age- and gender-matched healthy subjects were recruited. DTI-derived indices including fractional anisotropy (FA), mean diffusivity (MD), axial and radial diffusivities were quantified in the lateral and dorsal columns of cervical spinal cord. Based on the regions of interest (ROIs) measurement, NMO patients showed reduced FA, increased MD and radial diffusivity compared to control subjects, while axial diffusivity did not show any significant difference. The three former DTI metrics also showed significant correlations with disability scores, and especially FA was found to be sensitive to mild NMO. Our results show that DTI-derived indices can quantitatively assess the white matter (WM) abnormalities with seemingly normal appearance in conventional MRI, and are associated with the level of clinical disability, suggesting that DTI may have great potential as a useful diagnostic tool in the clinical setting.&#13;\n DKI is an extension of conventional DTI to probe the non-Gaussian diffusion property in biological tissues. Besides the four conventional DTI-derived metrics, DKI also provide three additional kurtosis metrics (mean kurtosis (MK), axial and radial kurtosis). In the second study, ROI-based analysis was used to characterize age-related microstructural changes in WM, cortical and subcortical gray matter (GM) of 27 healthy adults (21~59 yrs). Though the volumes of GM and WM were still preserved, DTI-derived metrics can detect the subtle changes in WM and GM. Meanwhile, MK and radial kurtosis significantly increased in both caudate nucleus and putamen while Thalamus showed little aging effect in the diffusivity and kurtosis metrics but significantly decreased only in FA. Our results demonstrated that DKI is sensitive to detect the age-related alterations in neural microstructures at the stage of early aging.&#13;\n In addition, DKI has been applied to detect the pathological changes in the normal-appearing neural tissues of 18 patients with multiple sclerosis (MS), compared to 22 healthy controls. Diffuse WM abnormalities have been observed extensively in the brain, revealed by DKI-derived metrics. Though the volumetric and voxel-wise analysis revealed no significant changes in the volume of cortical GM, decreased FA and kurtoses with increased diffusivities in MS group were sensitive to disclose the subtle alterations in global and regional cortical GM tissues. Significant correlations have been found between FA in the global, frontal and temporal cortical GM in relapsing-remitting MS patients and their disability scores, suggesting FA as an important biomarker to monitor the disease progress in cortical GM. Moreover, elevated kurtosis indices in MS patients did not correlate with diffusivities in caudate nucleus, putamen and thalamus, suggesting these metrics may be vulnerable to different pathologic aspects of the disease.&#13;\n In conclusion, DKI is sensitive to neural alterations during normal aging and in MS pathologies, and can provide complementary information to conventional MRI and DTI.

Traumatic brain injury (TBI) has been assessed with diffusion tensor imaging (DTI), a commonly used magnetic resonance imaging (MRI) marker for white matter integrity. However, given that the DTI model only fits a single fiber orientation, results can become confounded in regions of "crossing" white matter fibers. In contrast, constrained spherical deconvolution estimates a fiber orientation distribution directly from high angular resolution diffusion-weighted images. Consequently, constrained spherical deconvolution-based measures, such as apparent fiber density (AFD) and track-weighted imaging (TWI) metrics (including tract density imaging, average pathlength mapping, and mean curvature), may be more sensitive than DTI metrics to white matter injury post-TBI. As such, this study administered the lateral fluid percussion injury (FPI) model of TBI, assessed for changes in AFD and TWI metrics, and compared these results to the DTI metrics, fractional anisotropy (FA), mean diffusivity (MD), radial diffusivity (RD), and axial diffusivity (AD). Rats received either an FPI (n = 11) or sham injury (n = 9) and after a recovery period of 12 weeks underwent MRI. AFD was calculated as described previously and statistical testing was performed using connectivity-based fixel enhancement. TWI and DTI metrics were assessed using voxel-wise nonparametric permutation testing. We found that rats given an FPI had significantly reduced AFD, tract density, average pathlength, and mean curvature when compared to sham-injured rats and significant changes in DTI metrics, including reduced FA and increased MD, RD, and AD. However, the latter DTI metrics identified fewer voxels affected by TBI. Additionally, analysis of AFD with connectivity-based fixel enhancement was the only method that identified damage within the corticospinal tract of rats given an FPI. These results support the use of constrained spherical deconvolution, in conjunction with DTI metrics, to better assess disease progression and treatment post-TBI.

There are no studies to date,describing changes in the diffusion tensor imaging (DTI) metrics of the white matter (WM) regions of the entire cervical and thoracic spinal cord (SC) remote from the lesion in pediatric spinal cord injury (SCI) subjects. The purpose of this study was to determine whether DTI at sites cephalad and caudal to a lesion provides measures of cord abnormalities in children with chronic SCI. A retrospective study included 10 typically developing subjects (TD) and 10 subjects with chronic SCI who underwent SC imaging in 2014-2017. Axial diffusion tensor images using an inner field of view DTI sequence were acquired to cover the entire cervical and thoracic SC. Regions of interest were drawn on the SC WM: right and left lateral (motor), ventral (motor), and dorsal (sensory) tracts. To detect differences in DTI metrics between TD and SCI of the cord, a one way analysis of variance with pooled t test was performed. A stepwise regression analysis was performed to assess the correlation between DTI metrics and clinical scores. In motor and sensory tracts, fractional anisotropy (FA) and axial diffusivity (AD) were significantly decreased in the proximal segments of the caudal cord. In motor tracts cephalad to the lesion, FA was significantly decreased whereas AD was significantly increased in the proximal segment; however, AD was decreased in the distal and middle segments. International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI) total score was significantly correlated with FA and AD of the motor and sensory tracts cephalad to the lesion. This study demonstrates that FA and AD have the potential to be sensitive biomarkers of the full extent of cord injury and might be useful in detecting remote injuries to the SC and in guiding new treatments.

Correlation between spinal cord diffusion tensor imaging and postural response latencies in persons with multiple sclerosis: A pilot study

Tumors display chaotic vasculature that leads to hypoxia. Hypoxia is associated with an aggressive phenotype and increased resistance to radiation and chemotherapy. Hypoxic tumor regions also contain very few collagen 1 fibers, which is a major component of the tumor extracellular matrix (ECM) and plays an important role in molecular movement through the ECM. Here we examined the effects of hypoxia on water diffusion in a breast cancer xenograft model engineered to fluoresce under hypoxia. We observed, for the first time, that in hypoxic regions water diffusion and diffusion anisotropy was lower than in normoxic regions that may be explained by the few collagen fibers in hypoxic regions. Severe combined immunodeficient female mice were inoculated in the mammary fat pad with 2x106 MDA-MB-231 cells stably expressing red fluorescent protein (RFP) under the control of a hypoxia response element. Once tumor volume was approximately 300-400 mm3 the tumor was excised, fixed and imaged using high-resolution diffusion tensor imaging (DTI). The apparent diffusion coefficient (ADC) and fractional anisotropy (FA) maps were calculated. Following DTI, the tumor was sliced at 1 mm thick sections for optical imaging. Using a 1x lens to cover the entire tumor section, bright field and fluorescence optical images were acquired to detect the RFP-containing hypoxic regions. Multimodality registration was performed by feature extraction to co-register the optical images to the DTI images. Statistical dice similarity indices were calculated as a measure of the registration error. Hypoxic regions were identified from the optical fluorescence field images and the ADC and FA values were calculated in the corresponding hypoxic and normoxic regions. ADC and FA value distributions were identified in the DTI data and red fluorescent hypoxic regions were identified from the optical images. Co-registration accuracy confirmed by dice similarity index of 0.88 indicates a good overlap of optical and DTI images. Overall, hypoxic regions had lower water diffusion (ADC) and diffusion anisotropy (FA) values than normoxic regions. We have consistently observed that collagen fibers are significantly decreased in hypoxic regions, which could explain the lower ADC and FA values observed in hypoxic regions. In separate studies we have also observed high ADC in high collagen fiber containing regions in human breast cancer tissue. Low ADC and FA values observed in low collagen containing hypoxic regions indicate a functional role of these fibers in molecular transport. Decreased diffusion of molecules due to fewer collagen fibers in hypoxic regions may also contribute to poor drug delivery and tumor recurrence in hypoxic regions. Increased cellularity is thought to be one reason why lower ADC and FA values are observed in tumors. An even lower ADC within this backdrop may be due to hypoxic tumor regions. This work was supported by NIH P50CA103175 P30CA006973 Citation Format: Samata M. Kakkad, Jiangyang Zhang, Alireza Akhbardeh, Meiyappan Solaiyappan, Venu Raman, Dieter Leibfritz, Kristine Glunde, Zaver M. Bhujwalla. Water diffusion decreased in low collagen containing hypoxic regions of breast cancer xenograft. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 734. doi:10.1158/1538-7445.AM2013-734