The spleen is considered a non-essential organ. However, its importance is increasingly clear, given the serious disorders caused by its absence or dysfunction, e.g., greater susceptibility to infections, thromboembolism and cancer. Surgical techniques to preserve the spleen and maintain splenic function have become increasingly common. However, the morbidity and mortality associated with its absence and dysfunction are still high. We used the decellularization technique to obtain a viable splenic scaffold for recellularization
in vitro
and propose the idea of bioengineered spleen transplantation to the host. We observed the maintenance of important structural components such as white pulp, marginal zone and red pulp, in addition to the network of vascular ducts. The decellularized scaffold presents minimal residual DNA and SDS, which are essential to prevent immunogenic responses and transplantation failure. Also, the main components of the splenic matrix were preserved after decellularization, with retention of approximately 72% in the matrisomal protein content. The scaffold we developed was partially recellularized with stromal cells from the spleen of neonatal rats, demonstrating adhesion, proliferation and viability of cells. Therefore, the splenic scaffold is very promising for use in studies on spleen reconstruction and transplantation, with the aim of complete recovery of splenic function.
One of the main problems of the decellularization technique is the subjectivity of the final evaluation of its efficacy in individual organs. This problem can result in restricted cell repopulation reproducibility and worse responses to transplant tissues. Our proposal is to analyze the optical profiles produced by hearts during perfusion decellularization, as an additional method for evaluating the decellularization process of each individual organ. An apparatus comprised of a structured LED source and photo detector on an adjustable base was developed to capture the relationship between transmitted light during the perfusion of murine hearts, and residual DNA content. Voltage-time graphic records were used to identify a nonlinear mathematical model to discriminate between decellularizations with remaining DNA above (Incomplete Decellularization) and below (Complete Decellularization) the standardized limits. The results indicate that temporal optical evaluation of the process enables inefficient cell removal to be predicted in the initial stages, regardless of the apparent transparency of the organ. Our open system also creates new possibilities to add distinct photo detectors, such as for specific wavelengths, image acquisition, and physical-chemical evaluation of the scaffold, in order to collect different kinds of information, from dozens of studies. These data, when compiled and submitted to machine learning techniques, have the potential to initiate an exponential advance in tissue bioengineering research.
Euterpe edulis Martius is an endangered palm species that grows in the Atlantic Forest and the Cerrado of South America. Economic exploitation of its antioxidant-rich fruits could ensure the sustainable management of this species. However, this relies on the rapid selection of high-quality seeds from which to derive seedlings. The objective of this study was to investigate the maturation of E. edulis seeds using image analysis and X-ray densitometry. Fruits were harvested from ten matrices at different stages of maturation, from 94 days after anthesis (DAA) to 290 DAA. Seed dry mass, water content, germination, vigor, and density were quantified at each stage. At the same time, seeds were analyzed by GroundEye® imaging, radiography, scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS). The highest dry mass was detected 255 DAA (0.83 g), in vitro germination began 115 DAA and achieved 100% from 150 DAA, and maximum vigor was observed 164 DAA, whereby 100% of seedlings appeared normal. X-ray imaging revealed dehydrated seeds and small mechanical damage, such as cracking of the pericarp. X-ray densitometry revealed that seed density increased considerably 185 DAA. SEM/EDS detected changes between maturation stages, such as the accumulation of K and Si, in the mesocarp and endocarp. Overall, E. edulis seeds presented maximum in vitro germination, vigor, percentage of normal seedlings, and physicochemical qualities 164 DAA (green epicarp), which corresponds to 126 days earlier compared with the fruits harvested 290 DAA (black epicarp) for ex vitro germination.
Euterpe edulisMartius is an endangered species of the Atlantic Forest, whose fruits have high antioxidant potential, and propagated exclusively by seeds. The present study assessed the ability of different auxin inducers and picloram analogs to trigger somatic embryogenesis inE. edulis. Immature seeds were harvested, and their zygotic embryos were excised and grown in MS culture medium supplemented with 2,4-D dichlorophenoxyacetic acid (2,4-D) or picloram at 150, 300, 450, 600 µM. The activity of picloram analogs triclopyr and clopyralid was evaluated in semisolid MS medium. At maturation and germination, picloram-derived calli and somatic embryos isolated from triclopyr-grown cultures were first transferred to pre-maturation medium and, after 30 days, to basal MS or MS medium supplemented with either 5 µM abscisic acid or 0.53 µM 1-naphthaleneacetic acid plus 12.3 µM 2-isopentenyladenine. Finally, somatic embryos with root protrusions were transferred to MS medium devoid of sucrose for 30 days and then acclimatized ex vitro. Scanning, transmission, and atomic force microscopy revealed that picloram was superior to 2,4-D but less effective than triclopyr (100 µM) in inducing embryogenesis. Maturation and germination of somatic embryos in E. edulis can be maximized by 5 µM abscisic acid, and selecting calli via atomic force microscopy.HighlightThis work opens novel roads for embryogenic induction, using a new and more efficient inducer than the usual ones, and an innovative evaluation technique based on AFM.
The benefits of Kefir consumption are due, in part, to the rich composition of bioactive molecules released from fermentation. Angiotensin-converting enzyme (ACE) inhibitors are bioactive molecules with potential use in...
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