Análise da morfometria nuclear e textura da cromatina de amostras de carcinoma hepatocelular de pacientes transplantados hepáticos

Introdução: O carcinoma hepatocelular (CHC) é resultado de constantes lesões no fígado capazes de desencadear modificações na estrutura dos hepatócitos. A morfometria nuclear tem se mostrado útil na detecção destas alterações celulares, possibilitando que as mesmas sejam quantificadas, conferindo maior precisão e objetividade na análise histológica. Objetivo: Identificar alterações na estrutura nuclear e da textura de cromatina em amostras de CHC de pacientes submetidos ao transplante hepático. Materiais e métodos: Foram analisadas amostras de tecido tumoral e tecido adjacente não-neoplásico de 34 indivíduos submetidos à transplante hepático por CHC. Após a confirmação diagnóstica e a seleção da área exata do tumor, os tecidos foram reunidos em blocos de tissue microarray utilizados na confecção de lâminas histológicas coradas por Hematoxilina-Eosina. As imagens microscópicas de cada cilindro foram analisadas no software ImageJ quanto aos parâmetros nucleares de área, perímetro, circularidade, diâmetro de Feret, média de escala de cinza, solidez, razão de aspecto e dimensão fractal. Variáveis clinico-patológicas de relevância clínica foram obtidas através de consulta ao prontuário médico. Resultados: Dos 34 casos, 55,9% eram homens e 67,6% tinham história de hepatite C. Houve diferenças entre as amostras de CHC e tecido adjacente não-neoplásico quanto ao perímetro (p=0,025), circularidade (p=22,05 μm para perímetro com uma sensibilidade de 88% e especificidade de 85% (p=0,038). E de >1,14 μm para dimensão fractal com uma sensibilidade de 70% e especificidade de 91% (p =0,004). A correlação de Pearson inter-avaliadores teve significância (p=22.05 μm for perimeter with a sensitivity of 88% and specificity of 85% (p = 0.038). And of >1.14 μm for fractal dimension with a sensitivity of 70% and specificity of 91%(p=0.004). Pearson's inter-rater correlation was significant (p=<0.001) in relation to area, circularity, Feret's diameter, mean gray value and aspect ratio. Conclusions: A significant difference was found in the nuclear measurements (perimeter, circularity, solidity and aspect ratio) and in the chromatin texture between the samples of HCC and adjacent non-neoplastic tissue from liver transplant patients, as well as the association of these changes with relevant clinical characteristics (age, Model for End-Stage Liver Disease, recurrence and survival). The morphometric analysis showed high sensitivity and specificity regarding the perimeter and fractal dimension in the discrimination between neoplastic and non-neoplastic hepatocytes, with the ability to replicate inter-observer

Nuclear morphometry and chromatin texture changes in hepatocellular carcinoma samples may predict outcomes of liver transplanted patients

BACKGROUND: Nuclear changes are typical in the carcinogenesis of hepatocellular carcinoma (HCC). Morphometry and chromatin texture analysis are quantitative methods for their quantification. In this study, we analyzed nuclear morphometry and chromatin texture parameters in samples of hepatocellular carcinoma from liver transplant patients and their associations with clinicopathologic variables. METHODS: Samples of HCC and adjacent tissue from 34 individuals were collected in tissue microarray blocks. Stained slides were microphotographed using an optical microscope and nuclear parameters analyzed in ImageJ (FracLac plug-in). ROC curve analysis was used to find accurate cut-offs for differentiation of neoplastic and non-neoplastic cells. The inter-rater agreement was also evaluated. RESULTS: Nuclear morphometric and textural differences were observed between the samples of HCC and adjacent tissue of liver transplant patients. Lower mean gray value (p = 0.034) and Feret diameter (p = 0.024) were associated with higher Model for End-Stage Liver Disease (MELD) scores. Nuclei with larger area (p = 0.014) and larger Feret diameter (p = 0.035) were associated with lower survival. Lower aspect ratio was associated with HCC recurrence after the transplant (p = 0.048). The cut-off of 1.13 μm (p =  \u3c 0.001) for aspect ratio and cut-off of 21.15 μm (p = 0.038) for perimeter were established for the differentiation of neoplastic and non-neoplastic cells. The morphometric analysis was reproducible to area, circularity, Feret diameter, mean gray value and aspect ratio between observers (p =  \u3c 0.001). CONCLUSIONS: Nuclear morphometric differences between the HCC and the adjacent tissue samples were associated with prognostic variables (MELD scores, recurrence and survival) and may predict liver transplant patients\u27 outcomes

Nuclear morphometry and chromatin texture changes in hepatocellular carcinoma samples may predict outcomes of liver transplanted patients

Background: Nuclear changes are typical in the carcinogenesis of hepatocellular carcinoma (HCC). Morphometry and chromatin texture analysis are quantitative methods for their quantification. In this study, we analyzed nuclear morphometry and chromatin texture parameters in samples of hepatocellular carcinoma from liver transplant patients and their associations with clinicopathologic variables. Methods: Samples of HCC and adjacent tissue from 34 individuals were collected in tissue microarray blocks. Stained slides were microphotographed using an optical microscope and nuclear parameters analyzed in ImageJ (FracLac plug-in). ROC curve analysis was used to find accurate cut-offs for differentiation of neoplastic and non-neoplastic cells. The inter-rater agreement was also evaluated. Results: Nuclear morphometric and textural differences were observed between the samples of HCC and adjacent tissue of liver transplant patients. Lower mean gray value (p=0.034) and Feret diameter (p=0.024) were associated with higher Model for End-Stage Liver Disease (MELD) scores. Nuclei with larger area (p=0.014) and larger Feret diameter (p=0.035) were associated with lower survival. Lower aspect ratio was associated with HCC recurrence after the transplant (p=0.048). The cut-off of 1.13 μm (p= < 0.001) for aspect ratio and cut-off of 21.15 μm (p=0.038) for perimeter were established for the differentiation of neoplastic and non-neoplastic cells. The morphometric analysis was reproducible to area, circularity, Feret diameter, mean gray value and aspect ratio between observers (p= < 0.001). Conclusions: Nuclear morphometric differences between the HCC and the adjacent tissue samples were associated with prognostic variables (MELD scores, recurrence and survival) and may predict liver transplant patients’ outcomes