ObjectiveTo evaluate the ability of the eluate from fibrin-rich plasma (FRP) membrane to induce proliferation and differentiation of isolated human adipose-derived stem cells (ASCs) into chondrocytes.MethodFRP membranes were obtained by centrifugation of peripheral blood from two healthy donors, cut, and maintained in culture plate wells for 48 h to prepare the fibrin eluate. The SCATh were isolated from adipose tissue by collagenase digestion solution, and expanded in vitro. Cells were expanded and treated with DMEM-F12 culture, a commercial media for chondrogenic differentiation, and eluate from FRP membrane for three days, and labeled with BrdU for quantitative assessment of cell proliferation using the High-Content Operetta® imaging system. For the chondrogenic differentiation assay, the SCATh were grown in micromass for 21 days and stained with toluidine blue and aggrecan for qualitative evaluation by light microscopy. The statistical analysis was performed using ANOVA and Tukey's test.ResultsThere was a greater proliferation of cells treated with the eluate from FRP membrane compared to the other two treatments, where the ANOVA test showed significance (p < 0.001). The differentiation into chondrocytes was visualized by the presence of mucopolysaccharide in the matrix of the cells marked in blue toluidine and aggrecan.ConclusionsTreatment with eluate from FRP membrane stimulated cell proliferation and induced differentiation of the stem cells into chondrocytes, suggesting a potential application of FRP membranes in hyaline cartilage regeneration therapies.

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The aim of this study was evaluate the effects of Bracken fern (BF) (Pteridium aquilinum (L.) Kuhn.) on biological systems. When consumed by animals can cause acute intoxication, hematuria, biochemistry alterations and cancer. To humans the toxicity is associated with its intake on contaminated ground water or milk and inhalation of its spores. In order to check the BF aqueous extract (AEB) deleterious effects on animals blood vessels system, chick embryo chorioallantoic membrane (CAM) was used. It were applying on CAM 0.1, 0.5, 1, 5, 10 e 15 µg/mL of AEB and saline as control. The angiogenesis was analyzed and the vascular density index (VDI) calculated. The CAM samples were prepared and stained with H&E to evaluation of microvessels, Masson's trichrome to characterize collagen and fibrin deposition and Picro-sirius used to evaluate collagen using polarized light. Also the morphological aspects of embryos were analysed. We observe on the results of neovascularization that AEB did not change significantly the number of vessels/mm², however, membranes treated with AEB (5 or 10 µg/ mL) exhibit opacity and tissue fibrosis, both signs of inflammation. Histological analysis with Masson's trichrome and picro-sirius on tissues exposed to AEB respectively has shown increased collagen fibers and presence of fibrilar collagen. The embryos exposed to concentrations of 5 or 10 μg/mL AEB, showed changes as poor face formation and poor closing of abdominal wall. The highest concentration of AEB (15 μg/mL) was lethal to embryos. Although significant effects on the CAM's vasculature has not observed, tissue aggression was detected, a desmoplasia (an extensive inflammatory signal triggered by tissue injury), changes caused on embryos as well as the presence of toxic substances in the AEB show us an important and deleterious pathway of this bracken fern extract on its intoxicants effects on humans and animals, and even cancer or the death of animals. ResumoO foco principal desse estudo foi avaliar os efeitos do extrato aquoso de Samambaia (EAS) (Pteridium aquilinum (L.) Kuhn) sobre sistemas biológicos. A ingestão das folhas de Samambaia pode provocar intoxicação aguda, hematúria, alterações bioquímicas e câncer se consumida por animais. Em humanos também pode provocar intoxicação ao ser ingerida com água ou leite contaminados e pela inalação de seus esporos. Considerando os efeitos danosos provocados pelo consumo dessa planta esse estudo analisou a ação do EAS sobre um sistema de vasos sanguíneos, a membrana corioalantóide (MCA) de embrião de galinha. Sobre a MCA foram aplicados 0,1, 0,5, 1, 5, 10 e 15 µg/mL de EAS, salina como controle. A angiogênese foi analisada e o índice de densidade vascular (IDV) calculado. Amostras da CAM foram coradas com H&E para avaliação de microvasos, Tricrômico de Masson para caracterização de colágeno e deposição de fibrina e Picro-sirius para analisar o colágeno através de luz polarizada. Os aspectos morfológicos dos embriões também foram analisados. Os resultados da neovascularização mos...

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