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

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

Diferenciação celular e o processo de engenharia tecidual do sistema urinário

A questão central para a engenharia tecidual é induzir as células a recapitularem o seu fenótipo normal quando integradas a um arcabouço manufaturado. O objetivo deste trabalho foi examinar as variações na diferenciação celular do músculo liso e urotélio durante as etapas de produção de tecidos urinários in vitro. Quinze espécimes cirúrgicos geraram as culturas analisadas e os fenótipos das populações foram verificados por citometria de fluxo, imunofluorescência e expressão gênica. Houve uma associação entre o cultivo em superfícies plásticas e alterações no fenótipo de ambos os tipos celulares, os quais foram restaurados, em alguma extensão, após sua semeadura em matrizes temporárias. Enquanto o músculo liso recuperou o seu marcador contrátil, o urotélio apresentou atributos de uma camada basal. O estudo destacou como a perda da diferenciação secundária à expansão in vitro representa um desafio para a formação de neotecidos funcionais.The key issue of tissue engineering is how to induce cells to recapitulate their normal phenotype when placed within a scaffold. The aim of this work was to examine the smooth muscle and urothelium differentiation changes during the steps of urinary tissue production in vitro. Fifteen surgical specimens generated the analyzed cultures and phenotypes of cell populations were monitored by flow cytometer, immunofluorescence and gene expression. For both cellular types, there was an association of cell culture on plastic with phenotypic changes, but they were regained to some extent after cell seeding onto scaffolds. Smooth muscle cells restored their contractile marker, while urothelium showed attributes of a basal layer. The study highlights how the loss of differentiation due to propagation of cells in vitro represents a challenge to development of full functional neotissue

El Pueblo : diario republicano de Valencia: El Pueblo : diario republicano de Valencia - Año XV Número 5924 - 1907 diciembre 9 (09/12/1907)

Propósito: Os avanços obtidos pela engenharia de tecidos propiciaram o desenvolvimento de substitutos biológicos para tecidos urinários permanentemente danificados. O objetivo deste estudo é gerar um modelo para investigar as etapas fundamentais da formação de uma mucosa urotelial in vitro. Materiais e Métodos: Quatro tipos de matrizes foram produzidos com uma mistura de gelatina e diferentes combinações de ácido hialurônico e gelatina. Elas foram caracterizadas com o objetivo de verificar o efeito da modificação na composição sobre propriedades físico- -químicas relacionadas com a interação celular. As células utilizadas no estudo foram extraídas de segmentos do sistema urinário descartados após cirurgias. Após o período de cultura, tanto as células uroteliais quanto células-tronco derivadas de tecido adiposo foram semeadas nas matrizes manufaturadas. Cotes histológicos avaliaram a integração das células às matrizes. O multipotencial de diferenciação das células-tronco e a expressão de citoqueratina 7 pelo urotélio também foram investigados. Resultados: A maior interação da heparina com as moléculas da gelatina provocou uma redução na densidade de cargas eletrostáticas das matrizes. Este efeito também se refletiu na hidrofilicidade da superfície e na capacidade de absorção das matrizes em que a heparina estava presente. As células-tronco e o urotélio expandidos em culturas primárias demonstraram capacidade de proliferar em todas as matrizes. As células-tronco confirmaram seu multipotencial de diferenciação in vitro. As células uroteliais mantiveram a expressão de citoqueratina 7 ao serem cultivadas nas matrizes. Conclusão: Todas as matrizes produzidas suportaram a adesão e proliferação celulares.Purpose: Advances in culture techniques and production of temporary matrices have allowed the development of biological substitutes to injured urinary tissues. The aim of this work is to generate a model to study the fundamental steps required to produce a urothelial mucosa in vitro. Materials and Methods: Four types of matrices were manufactured based on a mixture of gelatin and different combinations of heparin and hyaluronic acid. They were characterized in order to evaluate the effect of composition on physicochemical properties related to cellular integration. Adipocyte-derived stem cells and urothelium were isolated from urinary tissues and expanded in culture. Mesenchymal stem cells were investigated so as to verify their multilineage potential. Each cellular type was seeded in matrices and after two weeks of culture the specimens were histologically analyzed. Additionally, urothelial cells expression of cytokeratin 7 was tested. Results: The addition of heparin to scaffolds decreased their electrostatic charges density, surface hydrophilicity and absorption capacity. Both cellular types were expanded in primary cultures and grew in all matrix types. Adipocyte-derived stem cells confirmed their multilineage potential. Urothelial cells maintained the cytokeratin 7 expression when cultured in matrices. Conclusion: Despite differences in composition and physicochemical properties, all matrices were able to support cellular attachment and proliferation

Diferenciação celular e o processo de engenharia tecidual do sistema urinário

A questão central para a engenharia tecidual é induzir as células a recapitularem o seu fenótipo normal quando integradas a um arcabouço manufaturado. O objetivo deste trabalho foi examinar as variações na diferenciação celular do músculo liso e urotélio durante as etapas de produção de tecidos urinários in vitro. Quinze espécimes cirúrgicos geraram as culturas analisadas e os fenótipos das populações foram verificados por citometria de fluxo, imunofluorescência e expressão gênica. Houve uma associação entre o cultivo em superfícies plásticas e alterações no fenótipo de ambos os tipos celulares, os quais foram restaurados, em alguma extensão, após sua semeadura em matrizes temporárias. Enquanto o músculo liso recuperou o seu marcador contrátil, o urotélio apresentou atributos de uma camada basal. O estudo destacou como a perda da diferenciação secundária à expansão in vitro representa um desafio para a formação de neotecidos funcionais.The key issue of tissue engineering is how to induce cells to recapitulate their normal phenotype when placed within a scaffold. The aim of this work was to examine the smooth muscle and urothelium differentiation changes during the steps of urinary tissue production in vitro. Fifteen surgical specimens generated the analyzed cultures and phenotypes of cell populations were monitored by flow cytometer, immunofluorescence and gene expression. For both cellular types, there was an association of cell culture on plastic with phenotypic changes, but they were regained to some extent after cell seeding onto scaffolds. Smooth muscle cells restored their contractile marker, while urothelium showed attributes of a basal layer. The study highlights how the loss of differentiation due to propagation of cells in vitro represents a challenge to development of full functional neotissue