A modified hydrogel production protocol to decrease cellular content

Purpose: To analyze the cytotoxicity and cell in porcine-derived decellularized skin matrix. Methods: We analyzed the effect of multiple decellularization processes by histological analysis, DNA quantification, and flow cytometry. Subsequently, we analyzed the most appropriate hydrogel concentration to minimize cytotoxicity on fibroblast culture and to maximize cell proliferation. Results: After the fourth decellularization, the DNA quantification showed the lowest DNA concentration (< 50 ng/mg). Histological analysis showed no cell components in the hydrogel. Moreover, hematoxylin and eosin showed a heterogeneous structure of collagen fibers. The best hydrogel concentration ranged from 3 to 25%, and there was no significant difference between the 24 hours and seven days. Conclusions: The process of hydrogel production was effective for removing cells and DNA elements. The best hydrogel concentration ranged from 3 to 25%.

Sex differences on solid organ histological characteristics after brain death

PURPOSE:To investigate gender differences in the evolution of the inflammatory process in rats subjected to brain death (BD).METHODS:Adult Wistar rats were divided into three groups: female; ovariectomized female; and male rats. BD was induced using intracranial balloon inflation and confirmed by maximal pupil dilatation, apnea, absence of reflex, and drop of mean arterial pressure. Six hours after BD, histological evaluation was performed in lungs, heart, liver and kidneys, and levels of inflammatory proteins, estrogen, progesterone, and corticosterone were determined in plasma.RESULTS:In the lungs, females presented more leukocyte infiltration compared to males (p<0.01). Ovariectomized female rat lungs were more hemorrhagic compared to other groups (p<0.001). In the heart, females had higher leukocyte infiltration and tissue edema compared to males (p<0.05). In the liver and kidneys, there were no differences among groups. In female group estradiol and progesterone were sharply reduced 6 hours after BD (p<0.001) to values observed in ovariectomized females and males. Corticosterone levels were similar.CONCLUSIONS:Sex hormones influence the development of inflammation and the status of organs. The increased inflammation in lungs and heart of female rats might be associated with the acute reduction in female hormones triggered by BD.(AU)

Influence of brain death and associated trauma on solid organ histological characteristics / Influência da morte encefálica e do trauma associado nas características histológicas de órgãos sólidos

PURPOSE: To evaluate histopathological alterations triggered by brain death and associated trauma on different solid organs in rats. METHODS: Male Wistar rats (n=37) were anesthetized with isoflurane, intubated and mechanically ventilated. A trepanation was performed and a balloon catheter inserted into intracraninal cavity and rapidly inflated with saline to induce brain death. After induction, rats were monitored for 30, 180, and 360 min for hemodynamic parameters and exsanguinated from abdominal aorta. Heart, lung, liver, and kidney were removed and fixed in paraffin to evaluation of histological alterations (H&E). Sham-operated rats were trepanned only and used as control group. RESULTS: Brain dead rats showed a hemodynamic instability with hypertensive episode in the first minute after the induction followed by hypotension for approximately 1 h. Histological analyses showed that brain death induces vascular congestion in heart (p<0.05), and lung (p<0.05); lung alveolar edema (p=0.001), kidney tubular edema (p<0.05); and leukocyte infiltration in liver (p<0.05). CONCLUSIONS: Brain death induces hemodynamic instability associated with vascular changes in solid organs and compromises most severely the lungs. However, brain death associated trauma triggers important pathophysiological alterations in these organs.(AU)

OBJETIVO: Avaliar as alterações histopatológicas desencadeadas pela morte encefálica e pelo trauma associado em diferentes órgãos sólidos em ratos. MÉTODOS: Ratos Wistar machos (n=37) foram anestesiados com isoflurano, entubados e mecanicamente ventilados. Foi realizada trepanação e um cateter foi inserido na cavidade intracraniana e insuflado rapidamente para induzir morte encefálica. Após a indução, os ratos foram monitorados por 30, 180 e 360 min para parâmetros hemodinâmicos e exsanguinados pela aorta abdominal. Coração, pulmão, fígado e rim foram removidos e fixados em parafina para avaliação de alterações histológicas (H&E). Ratos falso-operados foram apenas trepanados e usados como grupo controle. RESULTADOS: Ratos com morte encefálica apresentaram instabilidade hemodinâmica com episódio hipertensivo no primeiro minuto após a indução seguido de hipotensão por aproximadamente 1 hora. Análises histológicas demonstraram que a morte encefálica induz congestão vascular no coração (p<0,05) e pulmão (p<0,05); edema alveolar (p=0,001); edema tubular (p<0,05); e infiltrado leucocitário no fígado (p<0,05). CONCLUSÕES: A morte encefálica induz instabilidade hemodinâmica associada com mudanças vasculares em órgãos sólidos e compromete mais severamente os pulmões. Contudo, o trauma associado à morte encefálica desencadeia importantes alterações fisiopatológicas naqueles órgãos.(AU)

New quantitative variables to measure postoperative pericardial adhesions: useful tools in experimental research / Novas variáveis quantitativas para avaliação das aderências pericárdicas: ferramentas úteis na pesquisa experimental

PURPOSE: To improve the measurement system, during a research for the prevention of adhesions, we explored the feasibility of introducing three continuous numeric variables to quantify the intensity of pericardial adhesions. METHODS: To validate these three new numeric variables - time spent to dissect the adhesions (Δt), the amount of sharp dissection (ShpD) and the adhesion's collagen area (ACA) - as useful tools in measuring the severity of pericardial adhesions, data from a randomized study on adhesion prevention enrolling twenty-four swine, were analysed. A statistical Spearman's test and regressions models were applied to verify the correlation and the relationship between the results of a standard severity score (SS) and Δt, between SS and ShpD used in adhesiolysis and, also, between SS and ACA. RESULTS: There was a statistically significant correlation between SS and Δt, between SS and ShpD, as well as between SS and ACA, all measured by the Spearman's test (r=0.897, r=0.932, r=0.66; p<0.01, respectively). Through a non-linear regression, an exponential relation of SS with ShpD (R²=0.915) and SS with Δt (R²= 0.917) was found. CONCLUSION: The time spent to dissect the adhesions (Δt) and the amount of sharp dissection (ShpD) are new powerful measurement tools in evaluating outcomes of the methods used to prevent pericardial adhesions.(AU)

OBJETIVO: A fim de melhorar a metodologia de mensuração das aderências pericárdicas, avaliamos, durante um estudo de prevenção de aderências pericárdicas, a possibilidade da utilização de três de novas variáveis contínuas e numéricas. MÉTODOS: Para validar estas novas variáveis numéricas - tempo de dissecção (Δt), quantidade de dissecção cruenta (ShpD) e a área de colágeno na aderência (ACA) como instrumentos precisos na quantificação das aderências pericárdicas, foram analisados os dados de um estudo para prevenção de aderências que envolveu 24 porcos. Foram aplicados modelos de regressão e o teste de Spearman para avaliar a força e os tipos correlações entre os resultados do escore padrão de classificação de aderências (SS) e o Δt, entre o SS e o ShpD, e, também, entre o SS e a ACA. RESULTADOS: Foram evidenciadas correlações, estatisticamente significativas, entre o SS e o Δt, entre o SS e o ShpD, também como entre o SS e a ACA avaliadas através do teste de Spearman (r=0,897, r=0,932 , r=0,66; p<0,001, respectivamente). Identificou-se, através de regreção não-linear, uma relação exponencial do SS com o ShpD (R²=0,915) e com o Δt (R²= 0,917). CONCLUSÃO: Este estudo mostrou que o Δt e o ShpD são variáveis poderosas para avaliar os resultados dos métodos utilizados para prevenção das aderências pericárdicas.(AU)