Dott. Ing. Letizia Squarcina, Ph.D.

Post Doctoral Fellow

Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico

University of Milan
Scientific Institute IRCCS “E. Medea”, Bosisio Parini, Italy.

NEUROSCIENCE AND PSYCHIATRY

Tecniche di analisi di MRI cerebrale
 Diffusion MR images

Water diffusion is a random molecular motion (Brownian motion).

r = average diffusion distance
Einstein’s Law: <r2>= 2 D Td Td = diffusion time
D = diffusion coefficient
 Diffusion MR images
D= diffusion coefficient

Depends on:
- temperature
- type of medium
- molecular size

For example: free water at 37 °C = 3 x 10–9 m2 s-1

In tissues, water D is lower due to boundaries and obstacles (cellular

membranes, macromolecules, fibers etc)

D Le Bihan, Nat Rev Neurosc 4, 469-480 (2003)

 Diffusion MR images

DWI and DTI are MR techniques based on water diffusion properties.

Signal intensity is related to the random movement of water molecules, thanks to ad-hoc RF
pulses, applied before and after a RF pulse.

Interactions of water with intra- and extracellular spaces have an influence on water diffusion,
creating a perturbation of the random Brownian motion.

Measuring diffusion then can be useful to infer tissue properties which have an impact on
diffusion, e.g. tissue and cell integrity, both in normal subjects and in pathologies.
 Diffusion MR images
If water molecules are free to move in any direction, the distribution of their displacement has a
Gaussian shape, with 0 average.
1  x 2 / 2 2 1  x 2 / 4 Dt
P( x, t )  e  e
 2 4  D  t

At 37° C, after t=50 ms, 32% of molecules had a displacement of 17 mm, and only 5% moved of more than 34
mm.
 Diffusion MR images

Net displacement in Δt = 0 Net displacement in Δt ≠ 0

In tissues, water diffusion can be isotropic or anisotropic.

In isotropic diffusion, water molecules move in a random fashion, causing a net zero displacement. This
happens in tissues without organised structures, e.g. cerebro-spinal fluid.

When there are organised structures, e.g. white matter axon bundles, water is constrained to diffuse
following a preferential direction. In this case, the net displacement of water is different from zero.
Diffusion MR images

D = constant

D xx D xy D yx 
 
D   D yx D yy D yz 
 D zx D zy D zz 

 Diffusion MR images

Gray Matter

= =
Isotropic
MRI pixel unit Motion

= =
Anisotropic
White Matter Motion
 Diffusion MR images
Signal Generation

Gradient is applied along a direction, dependant on the position

Water molecules that move are subject to different values of B over time

Net phase shift

Signal attenuation

S = intensity of considered image voxel

S0 = non-diffusion-weighted image voxel intensity
S = S0e-bD D = diffusion tensor for considered voxel
b depends on sequence settings only

𝐷𝑥𝑥 𝐷𝑥𝑦 𝐷𝑥𝑧

𝐃 = 𝐷𝑦𝑥 𝐷𝑦𝑦 𝐷𝑦𝑧
𝐷𝑧𝑥 𝐷𝑧𝑦 𝐷𝑧𝑧
 Diffusion MR images
Diffusion tensor imaging (DTI) allows the investigation of brain structure and connections in vivo

A variety of indexes can be obtained, related to water diffusion in the brain, thus to brain structure

Water diffusion occurs predominantly along axons. DTI is most interesting in anisotropic tissues.

FA quantifies the anisotropy of diffusion: from 0 (sphere) to 1 (perfect anisotropy)

1 (unitless)

0
 Diffusion MR images
Diffusion tensor imaging (DTI) allows the investigation of brain structure and connections in vivo

A variety of indexes can be obtained, related to water diffusion in the brain, thus to brain structure

Water diffusion occurs predominantly along axons

MD indicates the magnitude of diffusion

0.01 mm2/s

0
 Diffusion MR images
Diffusion tensor imaging (DTI) allows the investigation of brain structure and connections in vivo

A variety of indexes can be obtained, related to water diffusion in the brain, thus to brain structure

Water diffusion occurs predominantly along axons

AD is related to diffusion happening the principal axis

0.004 mm2/s

0
 Diffusion MR images
Diffusion tensor imaging (DTI) allows the investigation of brain structure and connections in vivo

A variety of indexes can be obtained, related to water diffusion in the brain, thus to brain structure

Water diffusion occurs predominantly along axons

RD is related to diffusion happening perpendicularly to the principal axis

0.002 mm2/s

0
 Diffusion MR images
Diffusion tensor imaging (DTI) allows the investigation of brain structure and connections in vivo

A variety of indexes can be obtained, related to water diffusion in the brain, thus to brain structure

Water diffusion occurs predominantly along axons

VR gives a measure of the ratio between the ellipsoid volume and that of a
sphere which radius is equivalent to the mean diffusivity
1(unitless)

0
 Diffusion MR images
Diffusion tensor imaging (DTI) allows the investigation of brain structure and connections in vivo

A variety of indexes can be obtained, related to water diffusion in the brain, thus to brain structure

Water diffusion occurs predominantly along axons

MO1 is related to the three-dimensional characteristics of anisotropy, that

range from linear (1) to planar (-1)
1 (unitless)

1Ennisand Kindlmann, G (2006).

Orthogonal tensor invariants and
the analysis of diffusion tensor
-1 magnetic resonance images.
Magn Reson Med, 55, 1:136-46.
 Diffusion MR images
Colour FA Map

Demonstrates the direction of fibres

 Diffusion MR images

So, with DTI it is possible to estimate white matter tracts

 Diffusion MR images
Tractography - Overview

The data extracted from the imaging data is used to infer where fibre tracts are
Voxels are connected based upon similarities in the maximum diffusion direction

Johansen-Berg et al.
Ann Rev. Neurosci 32:75-94 (2009)
 Diffusion MR images

Degree of anisotropy Streamline tractography Probabilistic tractography

Nucifora et al. Radiology 245:2 (2007)

 Diffusion MR images
Deterministic tractography
Starting from a seed voxel (or ROI), every voxel along the path is connected to the adjacent following the principal direction of
diffusion.

The streamline points toward the next voxel and the tract is continued.

v3
v2 v1

Tracking termination criteria:

- FA falls below a certain value (usually around 0.2)
- The angle between two adjacent voxels is too wide
 Diffusion MR images

Probabilistic tractography

Tracts that originate from a seed ROI or voxel are represented with the probability of the presence of a fiber
bundle in the considered voxel.

Noise that inherently corrupts DTI data is considered and fully described.

Behrens et al, 2003

Behrens , T.E. , Woolrich, M.W., Jenkinson, M., Johansen-Berg, H., Nunes , R.G., Clare, S., Matthews, P.M., Brady, J.M., and Smith, S.M: Characterization and propagation
of uncertainty in diffusion-weighted MR imaging. Magn Reson Med, 50 (Nov 2003) 1077-1088.
 Diffusion MR images

Deterministic Tractography
Fast Doesn’t account for noise
Reliable in white matter Heavily dependent on FA value

Easy interpretation of results Doesn’t account for more than 1 direction

Probabilistic Tractography
Accounts for noise Computationally very heavy

Works also for low FA values Needs postprocessing

Multiple directions
 Diffusion MR images

TBSS on bipolar disorder and schizophrenia

Background and motivation
Changes in diffusion in bipolar disorder (BD) and schizophrenia (SCZ) have been
investigated, under the hypothesis that diffusion alterations can unveil structural brain
connectivity modifications characterizing major psychoses.

These two pathologies share common symptoms and modifications to brain structures1,2

Thus, we hypothesize he presence of common endophenotypes in SCZ and BD, which lead
for example to damage to corpus callosum and interhemispheric connectivity3,4

Dataset:
- 33 patients with DSM-IV BD (all Caucasians 48.9 +/- 8.3 years old; 25 females, 12 depressed, 3 hypomanic, 12
euthymic; and 1 in mixed state; 15 bipolar type I, 7 bipolar type II), 19 patients affected by SCZ (all Caucasians, 46.1
+/- 11.5 years old, 6 females) and 35 healthy controls (16 females 39 +/- 13 years old) were recruited

- DTI images (TR=8900 ms, TE=104 ms, 256x256x50 voxels, 0.92x0.92x3 mm3, b=1000 s/mm2). were acquired with a
1.5T Siemens Simphony (Siemens Healthcare, Erlangen, Germany) scanner along 11 directions.
 Diffusion MR images
In BOTH BD and SCZ

FA decreased
MO decreased
In respect to healthy controls
MD increased
VR increased
RD increased

Only in BD

AD increased

Changes in diffusion were widespread in the major white matter tracts, including

corpus callosum
corona radiata
longitudinal fasciculus
internal and external capsulae
thalamic radiations
 Diffusion MR images

We computed the areas where all considered diffusion indexes (except AD) showed changes in respect to healthy
simultaneously

White matter tracts areas where FA, MD, VR and RD show a significant change in patients affected by SCZ
 Diffusion MR images

We computed the areas where all considered diffusion indexes (except AD) showed changes in respect to healthy
simultaneously

White matter tracts areas where FA, MD, VR , RD and AD show a

significant change in patients affected by SCZ
 Diffusion MR images
Length of the disease significantly correlated (negatively) with FA, and (positively) with VR and RD (p=0.05, TFCE corrected) for
both populations of patients.

No other significant correlations were found.

Areas where FA negatively correlates with length of disease for SCZ

(p=0.05, TFCE corrected)
Affected voxels are concentrated in the body of corpus callosus
 Diffusion MR images
Length of the disease significantly correlated (negatively) with FA, and (positively) with VR and RD (p=0.05, TFCE
corrected) for both populations of patients.

No other significant correlations were found.

Areas where RD positively correlates with length of disease for SCZ

(p=0.05, TFCE corrected)
Affected voxels are concentrated in the body of corpus callosus
 Diffusion MR images
Length of the disease significantly correlated (negatively) with FA, and (positively) with VR and RD (p=0.05, TFCE corrected)
for both populations of patients.

No other significant correlations were found.

Areas where FA negatively correlates with length of disease for BD (p=0.05, TFCE corrected)
Affected voxels are concentrated in the in external capsule, corona radiata, corpus callosum and thalamic radiation.
 Diffusion MR images
Length of the disease significantly correlated (negatively) with FA, and (positively) with VR and RD (p=0.05, TFCE
corrected) for both populations of patients.

No other significant correlations were found.

Areas where RD positively correlates with length of disease for BD (p=0.05, TFCE corrected)
Affected voxels can be noted in the in anterior and posterior left and right corona radiata and right thalamic radiation.
 Diffusion MR images
Direct comparison between the two groups of patients, after correction for age and multiple comparisons, did not show
statistically relevant differences.

This demonstrates that changes underlying the pathophysiology of SCZ and BD are similar when structural connectivity is
considered.

Increases in AD as we found in BD patients in the right fronto-temporal network (corona radiata, superior longitudinal
fasciculus and internal capsule), are associated with axonal integrity

especially when coupled with an

increase in RD

water content is augmented outside the axons, mirroring a loss of white matter integrity
 Diffusion MR images
CONCLUSIONS

There are clusters of voxels where all considered diffusion indexes are altered, in more affected areas as the thalamo-
cortical radiations and corpus callosum

Interhemispheric connectivity is hindered in BD and SCZ, representing a common neural underpinning of major
psychoses. This confirms previous studies which described changes in corpus callosum in these pathologies, as already
demonstrated by our group1,2

1Brambilla et al (2009), White matter connectivity in bipolar disorder. Int Rev of Psychiatry 21: 380-386, 2009.
2Brambilla and Tansella (2007), The role of white matter for the pathophysiology of schizophrenia. International Review of Psychiatry 19: 459-68, 2007
 Diffusion MR images

TBI connectivity
Background and motivation

Traumatic brain injury (TBI) damages white matter pathways disrupting the functioning of
brain networks. The thalamus is a key node in many of these networks

It is difficult to study thalamic connectivity in vivo. DTI provides a flexible way of

investigating thalamo-cortical connections. In TBI patients, tractography often gives
uncertain results, because of the damage.

Dataset

22 TBI patients, 21 healthy controls

3T scanner
64 directions DTI sequence
128x128x73 voxel matrix

Squarcina et al., HBM 2012

 Diffusion MR images
TBI connectivity: methods
WM tracts
skeleton
• ROIs on T1 space Individual tractography
• Registration to DTI on healthy
Target ROI

• Tractography on healthy
Seed ROI

…
• White matter tracts
skeleton generation
Averaged outputs in
standard space

• Registration to atlas
• Averaged outputs
 Diffusion MR images
Healthy Patient
mean FA/MD FA/MD
• Generation of healthy mean FA and MD

Masked tract template

• Comparison of patients parameters with
healthy, inside the template:

Identification of abnormal voxels when:

MD > mean MD + 3sd

FA < mean FA – 3 sd

Voxels with pathological FA/MD

 Diffusion MR images
TBI connectivity: results
10 averaged tracts in MNI standard space were obtained: tracts from thalamus to anterior cingulate cortex, superior frontal
gyrus, inferior frontal gyrus, superior temporal gyrus, superior parietal lobe.
anterior cingulate superior frontal inferior frontal

superior parietal superior temporal

 Diffusion MR images
Using this template the evaluation of damage in the white matter can be done also in areas that would be discarded with an
individual tractography approach

healthy
low damage patient
high damage patient
 Diffusion MR images

Using the template, the number of voxels with abnormal diffusion characteristics was in general higher than using the output
of tractography, especially for patients with a high level of damage
80 20

60 15

Template
40 10
Tractography

20 5

0 0
accl accr ifgl ifgr sfgl sfgr spll splr stgl stgr accl accr ifgl ifgr sfgl sfgr spll splr stgl stgr
 Diffusion MR images

In this work, a novel approach for the evaluation of brain structure and integrity has been
proposed. In particular, a novel method for white matter tractography and a novel method
for the evaluation of structural thalamo-cortical connectivity have been presented.

The development of a novel tractography method focused on the evaluation of white

matter lesions has been demonstrated to highlight the pathologic process that disrupts
white matter connections.

With the proposed automated method for the investigation of thalamo-cortical

connectivity, the evaluation of white matter damage caused by trauma overcomes the
difficulties encountered by fiber tracking in areas with widespread damage.