Microwave assisted water extraction of hybrid carrageenan and antioxidant compounds from Mastocarpus stellatus red alga was first proposed to recover biopolymers with specific rheological properties, saving time, avoiding chemicals and enhancing extraction yields when compared with conventional treatments. The hybrid carrageenans, precipitated from the liquid fractions recovered after microwave assisted water extraction were analyzed by Fourier transform infrared attenuated total reflectance, high-performance size-exclusion chromatography and rheology. The corresponding soluble extracts were also characterized by total phenolic content, antioxidant capacity, proteins, sulfate and carbohydrate content. The highest phenolic, antioxidant capacity and sulfate content were observed at 190°C, without notable effect of heating time between 3 and 6 min. In contrast, a clear impact of both microwave heating temperature and time on the molecular weight distribution of hybrid carrageenans as well as on the viscoelastic behavior of formulated hydrogels was identified, being the strongest hydrogels achieved at 170°C for 6 min. It should be highlighted that all proposed hydrogels exhibited stable mechanical properties in the absence of syneresis for a month of cold storage, except those made with biopolymers obtained with the mildest extraction conditions (3 min at 70°C).
Glioblastoma is one of the most common and lethal types of primary brain tumor. Despite aggressive treatment with chemotherapy and radiotherapy, tumor recurrence within 6–9 months is common. To overcome this, more effective therapies targeting cancer cell stemness, invasion, metabolism, cell death resistance and the interactions of tumor cells with their surrounding microenvironment are required. In this study, we performed a systematic review of the molecular mechanisms that drive glioblastoma progression, which led to the identification of 65 drugs/inhibitors that we screened for their efficacy to kill patient-derived glioma stem cells in two dimensional (2D) cultures and patient-derived three dimensional (3D) glioblastoma explant organoids (GBOs). From the screening, we found a group of drugs that presented different selectivity on different patient-derived in vitro models. Moreover, we found that Costunolide, a TERT inhibitor, was effective in reducing the cell viability in vitro of both primary tumor models as well as tumor models pre-treated with chemotherapy and radiotherapy. These results present a novel workflow for screening a relatively large groups of drugs, whose results could lead to the identification of more personalized and effective treatment for recurrent glioblastoma.
Background Organoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months). Results Here we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids. Conclusions This new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.
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