| Summary: | INTRODUÇÃO: Imagens por tensores de difusão (diffusion tensor imaging - DTI) do cérebro são parte fundamental de exames de ressonância magnética (RM) na rotina clínica. Entretanto, não há informação científica suficiente para inferir a base histológica de parâmetros de imagens de DTI. Um dos achados frequentes nos exames de RM são as áreas de alteração de sinal na substância branca (AASSB). Neste estudo, analisamos imagens de RM post-mortem, em particular as AASSB, e seus correlatos histológicos e imuno-histoquímicos. OBJETIVOS: Descrever os parâmetros quantitativos de imagens de DTI imagens obtidas in cranio post-mortem, comparar com parâmetros de relaxometria e transferência de magnetização e avaliar seus correlatos histológicos nas AASSB e AHAN, utilizando técnicas de correspondência espacial ponto-a-ponto. MÉTODOS: Analisamos áreas distribuídas na substância branca de encéfalos de quatro sujeitos submetidos ao exame de RM post-mortem, aproximadamente 12,87 horas (±2,59). A RM foi realizada em sistema com campo de 3,0 T, sendo utilizadas imagens FLAIR; T2 multi-eco, DTI, Densidade de Prótons para cálculo da taxa de transferência de magnetização (magnetization transfer ratio - MTR). Foram delimitadas regiões de interesse (ROI) em 20 áreas contendo AASSB e 20 áreas homólogas aparentemente normais (AHAN). Para cada área, foram obtidos valores de anisotropia fracional (FA), coeficiente de difusão aparente (ADC), MTR e tempo T2. Os encéfalos foram seccionados em fatias de 0,4mm e fotografados em alta resolução. Estas imagens foram co-registradas tridimensionalmente com as imagens de RM, utilizando técnica semi-automatizada para correlação ponto-a-ponto. Os fragmentos analisados foram corados por hematoxilina-eosina (HE), Kluverx Barrera (KB), galocianina (GALL), anti-neurofilamento (anti-NF), antiproteína básica de mielina (anti-MBP) e anti-proteína ácida fibrilar glial (anti- GFAP). RESULTADOS: Os valores de FA foram 0,40 ±0,12 nas AASSB, e 0,52±0,15 nas AHAN. Os valores de ADC foram nas AASSB de 0,129±0,04 x 10-3mm2/s, e 0,108±0,03 x10-3mm2/s nas AHAN. Os valores de MTR foram 51,59±0,88 % nas AASSB, e 52,88±4,09 % nas AHAN. Os valores de RT2 foram 91,79±1,95 % nas AASSB, e 69,94±4,54 % nas AHAN. Os valores de HE foram 162,9±20,0 nas AASSB, e 163,3±20,4 nas AHAN. Os valores de GALL foram de 181,8±4,36 nas AASSB, e 167,11±20,65 nas AHAN. Os valores de KB foram de 168,9±19,08 nas AASSB, e 182.82±20,65 nas AHAN. Os valores de NF médios mensurados foram de 131,6±12,23 nas AASSB, e 128,03±4,74 nas AHAN. Os valores de MBP médios mensurados foram de 153,9 ±13,67 nas AASSB, e 153,18±12,69 nas AHAN. Os valores de GFAP médios mensurados foram de 94.61±7,85 nas AASSB, e 81,23±6,29 nas AHAN. Os testes não paramétricos de correlação (Spearman) entre variáveis de imagens de RM e histologia apresentaram correlação apenas entre FA e GFAP (r=-0,59; p < 0,005). DISCUSSÃO E CONCLUSÕES: De acordo com os resultados obtidos, podemos inferir que a presença de gliose é um processo patológico que pode ser detectado utilizando os parâmetros de FA nas imagens de DTI. Os resultados apresentados demonstram que imagens de RM adquiridas post-mortem podem servir para compreender melhor as alterações estruturais do tecido cerebral === BACKGROUND: Diffusion tensor imaging (DTI) is part of magnetic resonance imaging (MRI) acquisition methods available in clinical exams. However, there are not enough scientific basis to infer the histological substract of DTI parameters. White matter hyperintensities (WMH) are frequent findings in clinical MRI routine. In the last years, scientific evidences show that these injuries are not just age-related benign changes, but they could be associated to pathological processes. In this study we analyzed post-mortem MRI DTI data, specifically investigating WMH, their histological and immunohistochemical correlates. OBJECTIVES : At present study, we aimed to analyze quantifiable DTI parameters - Fractional Anisotropy (FA) and Apparent Diffusion Coefficient (ADC) - relaxometry (RT2) and magnetization transfer ratio (MTR) in WMH compared and normal appearing white matter (NAWM) from images obtained postmortem and in situ, and assess their histological substracts at WMH and NAWM using a point-topoint correlation platform. METHODS : We analyzed 20 regions of interest (ROI) encompassing WMH and NAWM selected from four subjects using in situ post-mortem MRI data acquired in 3.0T MR system. The subjects were scanned with a post-mortem interval of approximately 12h54m (±2h36m). The MRI analysis included 3D T1, FLAIR; multi-echo T2, DTI, and proton density to calculate the rate of magnetization transfer (magnetization transfer ratio - MTR). In each ROI we obtained measures of FA, ADC, RT2 and MTR. Brain specimens were posteriorly fixed in celloidin, sectioned into 0.4mm slices and photographed in high resolution. These images were co-registered with three-dimensional MR images using semi-automated technique for correlation using a point-to-point method established in our group. WMH and NAWM ROIs from MRI were submitted to quantitative histological analysis based on optical density color-deconvolution technique. Histological section were stained in hematoxylin-eosin (HE) , Kluver-Barrera (KB) , galloccyanin (GALL), anti-neurofilament (anti-NF) , anti-basic myelin protein (anti-MBP) and anti-glial fibrillary acidic protein (anti-GFAP). RESULTS: The mean FA values were 0.40±0.12 in WMH, and 0.52±0.15 in NAWM. ADC in WMH was 0.1290±0.04, x10-3mm2/s and 0.1081± 0.03 x10-3mm2/s in NAWM. MTR was 51.59± 0.88% in WMH, and 52.88±4.09% in NAWM. RT2 was 91.79±1.95% in WMH, and 69.94±4.54% in NAWM. FA, ADC and RT2 values were different between WMH and NAWM in a non-parametric analysis. On the quantitative histological analysis, the HE was 162.9±20.0 in WMH , and 163.3±20.4% in NAWM. GALL values at WMH was 181.8±4.36 and 167.11±20.65 at NAWM. KB was 168.9±19.08 in WMH, and 182.82±20.65 at NAWM. NF mean were 131.6 ±12.23 in WMH, and 128.03 ±14.74 at NAWM. MBP was 153.9±13.67 at WMH, and 153.18±12.69 at NAWM. Only GFAP values were different between WMH and NAWM. Mean GFAP was 94.61±7.85 in WMH, and 81.23±6.29 at NAWM. Non-parametric test (Spearman) between MR images and histology, showed correlation only for FA vs GFAP (r=-0.594, p < 0.005). DISCUSSION AND CONCLUSIONS: DTI FA and ADC parameters (and RT2 values) showed different patterns in WMH compared to NAWM, indicating that MR images acquired post-mortem may serve to understand the structural changes of the brain tissue. The presence of gliosis is a pathological process seems to be related to FA value from DTI
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