Soft tissues are commonly fiber-reinforced hydrogel composite structures, distinguishable from hard tissues by their low mineral and high water content. In this work, we proposed the development of 3D printed hydrogel constructs of the biopolymers chitosan (CHI) and cellulose nanofibers (CNFs), both without any chemical modification, which processing did not incorporate any chemical crosslinking. The unique mechanical properties of native cellulose nanofibers offer new strategies for the design of environmentally friendly high mechanical performance composites. In the here proposed 3D printed bioinspired CNF-filled CHI hydrogel biomaterials, the chitosan serves as a biocompatible matrix promoting cell growth with balanced hydrophilic properties, while the CNFs provide mechanical reinforcement to the CHI-based hydrogel. By means of extrusion-based printing (EBB), the design and development of 3D functional hydrogel scaffolds was achieved by using low concentrations of chitosan (2.0-3.0% (w/v)) and cellulose nanofibers (0.2-0.4% (w/v)). CHI/CNF printed hydrogels with good mechanical performance (Young's modulus 3.0 MPa, stress at break 1.5 MPa, and strain at break 75%), anisotropic microstructure and suitable biological response, were achieved. The CHI/CNF composition and processing parameters were optimized in terms of 3D printability, resolution, and quality of the constructs (microstructure and mechanical properties), resulting in good cell viability. This work allows expanding the library of the so far used biopolymer compositions for 3D printing of mechanically performant hydrogel constructs, purely based in the natural polymers chitosan and cellulose, offering new perspectives in the engineering of mechanically demanding hydrogel tissues like intervertebral disc (IVD), cartilage, meniscus, among others.Recently, the modification of the initial structure of biopolymers, mainly chitosan, has been gaining importance with a view to obtain functional forms with increased practicality and specific properties enabling their use in tissue engineering. https://www.selleckchem.com/products/smi-4a.html Therefore, in this article, the properties (structural and biological) of thermosensitive hydrogels obtained from chitosan lactate/chloride and two types of crosslinking agents (β-glycerol phosphate disodium salt pentahydrate and uridine 5'-monophosphate disodium salt) are discussed. The aim of the research is to identify changes in the structure of the biomaterials during conditioning in water. Structural investigations were carried out by FTIR spectroscopy. The crystallinity of gels was determined by X-ray diffraction analysis. The biocompatibility (evaluation of cytotoxicity and genotoxicity) of chitosan hydrogels was investigated by contact with human colon adenocarcinoma cell line for 48 h. The cytotoxicity was verified based on the colorimetric resazurin assay, and the genotoxicity was checked by the comet assay (percentage of DNA in the comet tail). The conducted research showed that the analyzed types of chitosan hydrogels are non-cytotoxic and non-genotoxic materials. The good biocompatibility of chitosan hydrogels surfaces makes them interesting scaffolds with clinical potential in tissue regeneration engineering.Accumulated lipid droplets in liver cause nonalcoholic fatty liver disease (NAFLD). Deep ocean water (DOW) containing high levels of magnesium, calcium, and potassium, etc. was proven to suppress hepatic lipid in obese rats fed high fat diet in the previous study. However, the effect of mineral compositions of DOW on the prevention of NAFLD is still unclear. This study removed calcium and potassium from DOW for modulating the mineral composition, and further compared the effects of DOW (D1(Mg + Ca + K)), DOW with low potassium (D2(Mg + Ca)), and DOW with low calcium and potassium (D3(Mg)) on the prevention of NAFLD in the **** model fed with high fat diet. In these results, DOW with high magnesium levels reduced serum and liver triglyceride and cholesterol levels and serum AST and ALT activities. However, when the calcium and/or potassium minerals were removed from DOW, the effects of reduction of triglyceride level, inhibition of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and peroxisome proliferator-activated receptor-alpha (PPAR-α) expressions, and activation of superoxide dismutase, catalase, and glutathione reductase activities would be weaker. In conclusion, DOW including magnesium, calcium and potassium minerals has the strongest preventive effect on NAFLD in a mouse model by increasing the antioxidant system and inhibiting fatty acid biosynthesis.The temperature range for consuming hot drinks includes temperatures that can damage cells on the tongue. We hypothesized that the consumption of very hot drinks can lead to a decrease in the ability to perceive low concentrations of tastants. We evaluated the ability to perceive low concentrations of five prototypical sapid compounds in 42 women and 40 men aged 18-65. A questionnaire made it possible to collect the usual frequencies (number of unit/day) and consumption temperature levels (medium hot/very hot) for four very common hot drinks (coffee, tea, herbal infusions, and hot chocolate). Our results showed that subjects who consumed very hot drinks (versus medium hot) were less sensitive to sweet (p = 0.020) and salty (p = 0.046) tastes. An aggravating effect of high consumption frequencies was only shown for sweet taste (p = 0.036). Moreover, our data also showed that women were more sensitive than men to sour, bitter, and umami tastes (p values less then 0.05), as well as that taste sensitivity decreases with age, especially after 50 years old (all tastes; p values less then 0.05). These findings strengthen our knowledge about the influence of sex and age on taste sensitivity, and they provide knowledge on the influence of consumption habits related to hot drinks on taste sensitivity.Patients with advanced-stage gastroenteropancreatic neuroendocrine tumors (GEP-NETs) have a poor overall prognosis despite chemotherapy and radiotherapy (e.g., peptide receptor radionuclide therapy (PRRT)). Better treatment options are needed to improve disease regression and patient survival. The purpose of this study was to examine a new treatment strategy by combining PI3K/mTOR dual inhibition and radiotherapy. First, we assessed the efficacy of two PI3K/mTOR dual inhibitors, PF-04691502 and PKI-402, to inhibit pAkt and increase apoptosis in NET cell lines (BON and QGP-1) and patient-derived tumor spheroids as single agents or combined with radiotherapy (XRT). Treatment with PF-04691502 decreased pAkt (Ser473) expression for up to 72 h compared with the control; in contrast, decreased pAkt expression was noted for less than 24 h with PKI-402. Simultaneous treatment with PF-04691502 and XRT did not induce apoptosis in NET cells; however, the addition of PF-04691502 48 h after XRT significantly increased apoptosis compared to PF-04691502 or XRT treatment alone.
Soft tissues are commonly fiber-reinforced hydrogel composite structures, distinguishable from hard tissues by their low mineral and high water content. In this work, we proposed the development of 3D printed hydrogel constructs of the biopolymers chitosan (CHI) and cellulose nanofibers (CNFs), both without any chemical modification, which processing did not incorporate any chemical crosslinking. The unique mechanical properties of native cellulose nanofibers offer new strategies for the design of environmentally friendly high mechanical performance composites. In the here proposed 3D printed bioinspired CNF-filled CHI hydrogel biomaterials, the chitosan serves as a biocompatible matrix promoting cell growth with balanced hydrophilic properties, while the CNFs provide mechanical reinforcement to the CHI-based hydrogel. By means of extrusion-based printing (EBB), the design and development of 3D functional hydrogel scaffolds was achieved by using low concentrations of chitosan (2.0-3.0% (w/v)) and cellulose nanofibers (0.2-0.4% (w/v)). CHI/CNF printed hydrogels with good mechanical performance (Young's modulus 3.0 MPa, stress at break 1.5 MPa, and strain at break 75%), anisotropic microstructure and suitable biological response, were achieved. The CHI/CNF composition and processing parameters were optimized in terms of 3D printability, resolution, and quality of the constructs (microstructure and mechanical properties), resulting in good cell viability. This work allows expanding the library of the so far used biopolymer compositions for 3D printing of mechanically performant hydrogel constructs, purely based in the natural polymers chitosan and cellulose, offering new perspectives in the engineering of mechanically demanding hydrogel tissues like intervertebral disc (IVD), cartilage, meniscus, among others.Recently, the modification of the initial structure of biopolymers, mainly chitosan, has been gaining importance with a view to obtain functional forms with increased practicality and specific properties enabling their use in tissue engineering. https://www.selleckchem.com/products/smi-4a.html Therefore, in this article, the properties (structural and biological) of thermosensitive hydrogels obtained from chitosan lactate/chloride and two types of crosslinking agents (β-glycerol phosphate disodium salt pentahydrate and uridine 5'-monophosphate disodium salt) are discussed. The aim of the research is to identify changes in the structure of the biomaterials during conditioning in water. Structural investigations were carried out by FTIR spectroscopy. The crystallinity of gels was determined by X-ray diffraction analysis. The biocompatibility (evaluation of cytotoxicity and genotoxicity) of chitosan hydrogels was investigated by contact with human colon adenocarcinoma cell line for 48 h. The cytotoxicity was verified based on the colorimetric resazurin assay, and the genotoxicity was checked by the comet assay (percentage of DNA in the comet tail). The conducted research showed that the analyzed types of chitosan hydrogels are non-cytotoxic and non-genotoxic materials. The good biocompatibility of chitosan hydrogels surfaces makes them interesting scaffolds with clinical potential in tissue regeneration engineering.Accumulated lipid droplets in liver cause nonalcoholic fatty liver disease (NAFLD). Deep ocean water (DOW) containing high levels of magnesium, calcium, and potassium, etc. was proven to suppress hepatic lipid in obese rats fed high fat diet in the previous study. However, the effect of mineral compositions of DOW on the prevention of NAFLD is still unclear. This study removed calcium and potassium from DOW for modulating the mineral composition, and further compared the effects of DOW (D1(Mg + Ca + K)), DOW with low potassium (D2(Mg + Ca)), and DOW with low calcium and potassium (D3(Mg)) on the prevention of NAFLD in the mice model fed with high fat diet. In these results, DOW with high magnesium levels reduced serum and liver triglyceride and cholesterol levels and serum AST and ALT activities. However, when the calcium and/or potassium minerals were removed from DOW, the effects of reduction of triglyceride level, inhibition of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and peroxisome proliferator-activated receptor-alpha (PPAR-α) expressions, and activation of superoxide dismutase, catalase, and glutathione reductase activities would be weaker. In conclusion, DOW including magnesium, calcium and potassium minerals has the strongest preventive effect on NAFLD in a mouse model by increasing the antioxidant system and inhibiting fatty acid biosynthesis.The temperature range for consuming hot drinks includes temperatures that can damage cells on the tongue. We hypothesized that the consumption of very hot drinks can lead to a decrease in the ability to perceive low concentrations of tastants. We evaluated the ability to perceive low concentrations of five prototypical sapid compounds in 42 women and 40 men aged 18-65. A questionnaire made it possible to collect the usual frequencies (number of unit/day) and consumption temperature levels (medium hot/very hot) for four very common hot drinks (coffee, tea, herbal infusions, and hot chocolate). Our results showed that subjects who consumed very hot drinks (versus medium hot) were less sensitive to sweet (p = 0.020) and salty (p = 0.046) tastes. An aggravating effect of high consumption frequencies was only shown for sweet taste (p = 0.036). Moreover, our data also showed that women were more sensitive than men to sour, bitter, and umami tastes (p values less then 0.05), as well as that taste sensitivity decreases with age, especially after 50 years old (all tastes; p values less then 0.05). These findings strengthen our knowledge about the influence of sex and age on taste sensitivity, and they provide knowledge on the influence of consumption habits related to hot drinks on taste sensitivity.Patients with advanced-stage gastroenteropancreatic neuroendocrine tumors (GEP-NETs) have a poor overall prognosis despite chemotherapy and radiotherapy (e.g., peptide receptor radionuclide therapy (PRRT)). Better treatment options are needed to improve disease regression and patient survival. The purpose of this study was to examine a new treatment strategy by combining PI3K/mTOR dual inhibition and radiotherapy. First, we assessed the efficacy of two PI3K/mTOR dual inhibitors, PF-04691502 and PKI-402, to inhibit pAkt and increase apoptosis in NET cell lines (BON and QGP-1) and patient-derived tumor spheroids as single agents or combined with radiotherapy (XRT). Treatment with PF-04691502 decreased pAkt (Ser473) expression for up to 72 h compared with the control; in contrast, decreased pAkt expression was noted for less than 24 h with PKI-402. Simultaneous treatment with PF-04691502 and XRT did not induce apoptosis in NET cells; however, the addition of PF-04691502 48 h after XRT significantly increased apoptosis compared to PF-04691502 or XRT treatment alone.
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