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Gray and White Matter Delineation in the Human Spinal Cord Using Diffusion Tensor Imaging and Fuzzy Logic

Rationale and Objectives

Diffusion tensor imaging (DTI) has been used extensively in determining morphology and connectivity of the brain; however, similar analysis in the spinal cord has proven difficult. The objective of this study was to improve the delineation of gray and white matter in the spinal cord by applying signal processing techniques to the eigenvalues of the diffusion tensor. Our approach involved creating anisotropy indices based on the difference between eigenvalues and mean diffusivity then using a fuzzy inference system (FIS) to delineate between gray and white matter in the human cervical spinal cord.

Materials and Methods

DTI was performed on the cervical spinal cord in five neurologically intact subjects. Distributions were extracted for regions of gray and white matter through the use of a digitized histologic template. Fuzzy membership functions were created based on these distributions. Detectability index and receiver operating characteristic (ROC) analysis was performed on traditional DTI indices and FIS classified regions.

Results

A significantly higher contrast between gray and white matter was observed using fuzzy classification compared with traditionally used DTI indices based on the detectability index ( P < .001) and trends in the ROC analysis. Reconstructed images from the FIS qualitatively showed a better anatomical representation of the spinal cord compared with traditionally used DTI indices.

Conclusions

Diffusion tensor imaging using an FIS for tissue classification provides high contrast between spinal gray and white matter compared with traditional DTI indices and may provide a noninvasive technique to quantify the integrity and morphology of the human spinal cord following injury.

Diffusion tensor imaging (DTI) is a magnetic resonance imaging method that quantifies the diffusion of water within a voxel in three-dimensional space. Indices of the diffusion tensor have been used for distinguishing between white and gray matter in the brain ( ) for the identification of brain structures, especially in pathologic circumstances. Similarly, classification of gray and white matter in the human spinal cord would be useful for characterizing morphologic features of the cord; however, spatial resolution restrictions and other challenges have limited the application of DTI in the spinal cord ( ). Although previous studies have quantified the basic diffusion properties of the human spinal cord using indices derived from the diffusion tensor ( ), delineation between gray and white matter in the in vivo spinal cord has not been accomplished. The purpose of this study was to develop an improved method for the delineation of gray and white matter in the human spinal cord through the use of a fuzzy logic–based tissue classification algorithm.

Previous research in the brain has shown that the diffusion of water is different for voxels containing white matter compared with those containing gray matter, most likely because of membrane barriers associated with the axons ( ) along with other morphologic and orientation-dependent characteristics ( ). Diffusion perpendicular to axon orientation is restricted by these boundaries causing white matter to be more anisotropic than gray matter; therefore, measures of anisotropy have been used for delineation of gray and white matter in the brain. Unfortunately, differences in diffusion properties of gray and white matter in the spinal cord are not as distinct, although gray matter is slightly more isotropic than white matter ( ). As a result, traditional anisotropy measures are not as effective for delineation of spinal gray and white matter.

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Materials and methods

Subjects

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MRI

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Anisotropy Indices

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ψ1(x,y,z)=λ1(x,y,z)−MD(x,y,z) ψ

1

(

x

,

y

,

z

)

=

λ

1

(

x

,

y

,

z

)

M

D

(

x

,

y

,

z

)

ψ2(x,y,z)=λ2(x,y,z)−MD(x,y,z) ψ

2

(

x

,

y

,

z

)

=

λ

2

(

x

,

y

,

z

)

M

D

(

x

,

y

,

z

)

ψ3(x,y,z)=λ3(x,y,z)−MD(x,y,z) ψ

3

(

x

,

y

,

z

)

=

λ

3

(

x

,

y

,

z

)

M

D

(

x

,

y

,

z

)

where ψ n is the anisotropy (mm 2 /second) in a specific direction n , λ n is the eigenvalue for a particular direction (mm 2 /second), and MD is the mean diffusivity, defined as the average of all three eigenvalues (mm 2 /second).

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Template Creation and Training Data

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Figure 1, Creation of a template from a histological slice of cervical spinal cord at the C2 level. The histologic image was obtained from Bergman et al ( 12 ). G = gray matter; W = white matter, P = partial volume between white matter and cerebrospinal fluid, counted as both white matter and cerebrospinal fluid. The template was used to generalize the morphology of the upper cervical spinal cord.

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FIS

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μ(χ)=[μCSF(χ)μGM(χ)μWM(χ)]=⎡⎣⎢μCSF(ψ1)μCSF(ψ2)μCSF(ψ3)μGM(ψ1)μGM(ψ2)μGM(ψ3)μWM(ψ1)μWM(ψ2)μWM(ψ3)⎤⎦⎥ μ

(

χ

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3

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μ

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M

(

ψ

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μ

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M

(

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where

μtissue(index)=12πsindex,tissue√e−(index−mindex,tissue)22⋅sindex,tissue μ

t

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is the Gaussian membership function for a specific anisotropy index and tissue type, m index,tissue is the sampled mean value for a particular index and tissue type obtained with the histologic template, and s index,tissue is the sampled standard deviation.

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β=and(μ(χ))=[and(μCSF(χ))and(μGM(χ))and(μWM(χ))] β

=

a

n

d

(

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and(μtissue(χ))=∏∈indexμtissue(index) a

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The degree of membership of a specific voxel as a particular tissue type was then mapped to the new variable space termed the fuzzy anisotropy index by first defining a set of output membership functions. To allow for a specific voxel being a combination of tissue types, as occurs with partial volume effects, we defined an output membership function matrix as

ξ(y)=[ξCSF(y)ξGM(y)ξWM(y)] ξ

(

y

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=

[

ξ

C

S

F

(

y

)

ξ

G

M

(

y

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ξ

W

M

(

y

)

]

where

ξCSF(y)={−2y+10for0≤y≤0.5for0.5≤y≤1 ξ

C

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(

y

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=

{

2

y

+

1

0

for

0

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ξGM(y)={2y−2y−2for0≤y≤0.5for0.5≤y≤1 ξ

G

M

(

y

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=

{

2

y

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for

0

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0

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ξWM(y)={02y−1for0≤y≤0.5for0.5≤y≤1 ξ

W

M

(

y

)

=

{

0

2

y

1

for

0

y

0

.5

for

0

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y

1

where y was the output variable termed the fuzzy anisotropy . To perform the mapping operation, the minimum between the membership grade for each tissue type, β, and the output membership function for that particular tissue type, ξ ( y ), was calculated. The membership grade for each tissue type was assumed constant for all values of the fuzzy anisotropy , y , thus β ( y ) = β. The mapped distributions were defined as

η(y)=[ηCSF(y)ηGM(y)ηWM(y)]=min(β,ξ(y)) η

(

y

)

=

[

η

C

S

F

(

y

)

η

G

M

(

y

)

η

W

M

(

y

)

]

=

min

(

β

,

ξ

(

y

)

)

where

ηtissue(y)=min(and(μtissue(χ)),ξtissue(y)) η

t

i

s

s

u

e

(

y

)

=

min

(

a

n

d

(

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s

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u

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(

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,

ξ

t

i

s

s

u

e

(

y

)

)

The result of the mapping operation was a row vector, η ( y ), containing three functions of the fuzzy anisotropy for each tissue type, η tissue ( y ). A single value for the fuzzy anisotropy index (FAI) was computed by first summing the three distributions in η ( y ) and then finding the centroid location along y . The FAI was defined as

FAI=centroid(η(y)φ) F

A

I

=

c

e

n

t

r

o

i

d

(

η

(

y

)

φ

)

where φ = [1 1 1] T was used as the matrix sum. The resulting FAI was used to classify gray matter regions and intact white matter tracts. The Fuzzy Logic Toolbox in MATLAB (MathWorks, Inc, Natick, MA) was used for implementation of the FIS.

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Detectability Index

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d=|mGM−mWM|s2GM+s2WM√. d

=

|

m

G

M

m

W

M

|

s

G

M

2

+

s

W

M

2

.

For the 15 test slices, m GM was the mean sampled index value for the GM region of interest, m WM was the mean sampled index value for the WM region of interest, s 2 GM was the sampled variance of a particular index for the GM region of interest, and s 2 WM was the sampled variance of a particular index for the WM region of interest. The detectability index was calculated for the traditional DTI indices, anisotropy indices ψ 1 , ψ 2 , ψ 3 , and the FAI from the FIS classification. A two-way analysis of variance with pooled variance was performed comparing the detectability index between DTI indices and subjects. Post-hoc statistical analysis was completed using Tukey’s test for multiple comparisons with α = 0.05. All statistical tests were completed with MINITAB statistical software.

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Receiver Operator Characteristic

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Results

Anisotropy Indices

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Figure 2, Traditional diffusion tensor imaging indices used in the spinal cord ( lADC , tADC , FA , MD ) for four slices of the cervical spinal cord in one subject (C1–C4) are shown. Vertebral levels were determined by sagittal locator images and anatomical landmarks. For illustration and comparison purposes, image resolution was artificially increased by a factor of four and linearly interpolated to create a smoothed image.

Figure 3, Anisotropy indices based on the difference between the eigenvalues and mean diffusivity (ψ 1 , ψ 2 , ψ 3 ) for four slices of the cervical spinal cord in one subject (C1–C4) are shown. For illustration and comparison purposes, image resolution was artificially increased by a factor of four and linearly interpolated to create a smoothed image.

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Template Alignment

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Figure 4, The percent area overlap between histologic template and cervical spinal cord images for five slices of the cervical spine for each subject ( n = 5) is shown. Error bars represent the standard deviation.

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FIS

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Figure 5, Distributions for anisotropy indices (ψ 1 , ψ 2 , ψ 3 ) used as membership functions for the fuzzy inference system (FIS) extracted using the histologic template are shown. Note that the cerebrospinal fluid was less anisotropic than both white and gray matter. Error bars represent the standard deviation.

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Figure 6, The fuzzy anisotropy index ( FAI ) and fractional anisotropy ( FA ) for four slices of the cervical spinal cord in one subject (C1–C4) is displayed. For illustration and comparison purposes, image resolution was artificially increased by a factor of four and linearly interpolated to create a smoothed image.

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Table 1

Percentage of Test Image Voxels Labeled Correctly for the Fuzzy Anisotropy Index and Fractional Anisotropy with Respect to Template Classification

Index Region % Correct n Fuzzy anisotropy index Gray matter 86.5 27 Fractional anisotropy Gray matter 70.4 27 Fuzzy anisotropy index White matter 67.1 60 Fractional anisotropy White matter 40.0 60

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Detectability Index

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Figure 7, The detectability index, d , between gray and white matter for traditional diffusion tensor imaging indices, newly developed anisotropy indices, and fuzzy anisotropy index using histological template for classification ( n = 15) is shown.

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Figure 8, Comparison of (a) fractional anisotropy ( FA ) to (b) longitudinal anisotropy, ψ 1 , and (c) fuzzy anisotropy index ( FAI ) at the C1 level is demonstrated. Note that the images in (b) and (c) illustrate features of the ventral horns (white arrows), which are lacking in (a) , the FA image. For illustration and comparison purposes, image resolution was artificially increased by a factor of four by zero padding and linear interpolation to create a smoothed image. Posterior direction is toward the top of the image, anterior direction is toward the bottom, left direction is to the right of the image, and right direction is to the left of the image.

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ROC

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Figure 9, Receiver operator characteristic (ROC) curve (a) showing sensitivity and specificity for lADC, tADC, FA, MD, ψ 1 , and fuzzy anisotropy index ( FAI ). (b) Area under ROC curve across subjects and diffusion tensor imaging indices.

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Discussion

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Imaging Sequence

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Traditional versus New Indices

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FIS

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Limitations of the Study

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Future Applications

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Conclusion

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Acknowledgments

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