Fabrication of Electrospun Double Layered Biomimetic Collagen-Chitosan Polymeric Membranes with Zinc-Doped Mesoporous Bioactive Glass Additives.

Several therapeutic approaches have been developed to promote bone regeneration, including guided bone regeneration (GBR), where barrier membranes play a crucial role in segregating soft tissue and facilitating bone growth. This study emphasizes the importance of considering specific tissue requirements in the design of materials for tissue regeneration, with a focus on the development of a double-layered membrane to mimic both soft and hard tissues within the context of GBR. The hard tissue-facing layer comprises collagen and zinc-doped bioactive glass to support bone tissue regeneration, while the soft tissue-facing layer combines collagen and chitosan. The electrospinning technique was employed to achieve the production of nanofibers resembling extracellular matrix fibers. The production of nano-sized (~116 nm) bioactive glasses was achieved by microemulsion assisted sol-gel method. The bioactive glass-containing layers developed hydroxyapatite on their surfaces starting from the first week of simulated body fluid (SBF) immersion, demonstrating that the membranes possessed favorable bioactivity properties. Moreover, all membranes exhibited distinct degradation behaviors in various mediums. However, weight loss exceeding 50% was observed in all tested samples after four weeks in both SBF and phosphate-buffered saline (PBS). The double-layered membranes were also subjected to mechanical testing, revealing a tensile strength of approximately 4 MPa. The double-layered membranes containing zinc-doped bioactive glass demonstrated cell viability of over 70% across all tested concentrations (0.2, 0.1, and 0.02 g/mL), confirming the excellent biocompatibility of the membranes. The fabricated polymer bioactive glass composite double-layered membranes are strong candidates with the potential to be utilized in tissue engineering applications.

Green Electrospun Poly(vinyl alcohol)/Gelatin-Based Nanofibrous Membrane by Incorporating 45S5 Bioglass Nanoparticles and Urea for Wound Dressing Applications: Characterization and In Vitro and In Vivo Evaluations.

This study reports the fabrication and characterization of poly(vinyl alcohol) (PVA) and gelatin (Gel)-based nanofiber membranes cross-linked with citric acid (CA) by a green electrospinning method in which nano 45S5 bioglass (BG) and urea were incorporated. Various combinations of PVA, gelatin, and BG were prepared, and nanofiber membranes with average fiber diameters between 238 and 595 nm were fabricated. Morphological, chemical, and mechanical properties, porosity, swelling, water retention, and water vapor transmission rate of the fabricated membranes were evaluated. PVA:Gel (90:10), 15% CA, and 3% BG were determined as the optimum blend for nanofiber membrane fabrication via electrospinning. The membrane obtained using this blend was further functionalized with 10% w/w polymer urea coating by the electrospray method following the cross-linking. In vitro biocompatibility tests revealed that the fabricated membranes were all biocompatible except for the one that functionalized with urea. In vivo macroscopic and histopathological analysis results of PVA/Gel/BG and PVA/Gel/BG/Urea treated wounds indicated increased collagenization and vascularization and had an anti-inflammatory effect. Furthermore, careful examination of the in vivo macroscopic results of the PVA/Gel/BG/Urea membrane indicated its potential to decrease uneven scar formation. In conclusion, developed PVA/Gel/BG and PVA/Gel/BG/Urea electrospun membranes with multifunctional and biomimetic features may have the potential to be used as beneficial wound dressings.

Evaluation of biocomposite putty with strontium and zinc co-doped 45S5 bioactive glass and sodium hyaluronate.

The application of powder or granule formed bioactive glasses in the defect area with the help of a liquid carrier to fill the defects is a subject of interest and is still open to development. In this study, it was aimed to prepare biocomposites of bioactive glasses incorporating different co-dopants with a carrier biopolymer and to create a fluidic material (Sr and Zn co-doped 45S5 bioactive glassessodium hyaluronate). All biocomposite samples were pseudoplastic fluid type, which may be suitable for defect filling and had excellent bioactivity behaviors confirmed by FTIR, SEM-EDS and XRD. Biocomposites with Sr and Zn co-doped bioactive glass had higher bioactivity considering the crystallinity of hydroxyapatite formations compared to biocomposite with undoped bioactive glasses. Biocomposites with high bioactive glass content had hydroxyapatite formations with higher crystallinity compared to biocomposites with low bioactive glass. Furthermore, all biocomposite samples showed non-cytotoxic effect on the L929 cells up to a certain concentration. However, biocomposites with undoped bioactive glass showed cytotoxic effects at lower concentrations compared to biocomposites with co-doped bioactive glass. Thus, biocomposite putties utilizing Sr and Zn co-doped bioactive glasses may be advantageous for orthopedic applications due to their specified rheological, bioactivity, and biocompatibility properties.

Anaerobic Glycolysis Maintains the Glomerular Filtration Barrier Independent of Mitochondrial Metabolism and Dynamics.

The cellular responses induced by mitochondrial dysfunction remain elusive. Intrigued by the lack of almost any glomerular phenotype in patients with profound renal ischemia, we comprehensively investigated the primary sources of energy of glomerular podocytes. Combining functional measurements of oxygen consumption rates, glomerular metabolite analysis, and determination of mitochondrial density of podocytes in vivo, we demonstrate that anaerobic glycolysis and fermentation of glucose to lactate represent the key energy source of podocytes. Under physiological conditions, we could detect neither a developmental nor late-onset pathological phenotype in podocytes with impaired mitochondrial biogenesis machinery, defective mitochondrial fusion-fission apparatus, or reduced mtDNA stability and transcription caused by podocyte-specific deletion of Pgc-1α, Drp1, or Tfam, respectively. Anaerobic glycolysis represents the predominant metabolic pathway of podocytes. These findings offer a strategy to therapeutically interfere with the enhanced podocyte metabolism in various progressive kidney diseases, such as diabetic nephropathy or focal segmental glomerulosclerosis (FSGS).

Hybrid cells derived from breast epithelial cell/breast cancer cell fusion events show a differential RAF-AKT crosstalk.

BACKGROUND: The biological phenomenon of cell fusion has been linked to several characteristics of tumour progression, including an enhanced metastatogenic capacity and an enhanced drug resistance of hybrid cells. We demonstrated recently that M13SV1-EGFP-Neo breast epithelial cells exhibiting stem cell characteristics spontaneously fused with MDA-MB-435-Hyg breast cancer cells, thereby giving rise to stable M13MDA435 hybrid cells, which are characterised by a unique gene expression profile and migratory behaviour. Here we investigated the involvement of the PLC-ß/γ1, PI3K/AKT and RAS-RAF-ERK signal transduction cascades in the EGF and SDF-1α induced migration of two M13MDA435 hybrid cell clones in comparison to their parental cells. RESULTS: Analysis of the migratory behaviour by using the three-dimensional collagen matrix migration assay showed that M13SV1-EGFP-Neo cells as well as M13MDA435 hybrid cells, but not the breast cancer cell line, responded to EGF stimulation with an increased locomotory activity. By contrast, SDF-1α solely stimulated the migration of M13SV1-EGFP-Neo cells, whereas the migratory activity of the other cell lines was blocked. Analysis of signal transduction cascades revealed a putative differential RAF-AKT crosstalk in M13MDA435-1 and -3 hybrid cell clones. The PI3K inhibitor Ly294002 effectively blocked the EGF induced migration of M13MDA435-3 hybrid cells, whereas the EGF induced locomotion of M13MDA435-1 hybrid cells was markedly increased. Analysis of RAF-1 S259 phosphorylation, being a major mediator of the negative regulation of RAF-1 by AKT, showed decreased pRAF-1 S259 levels in LY294002 treated M13MDA435-1 hybrid cells. By contrast, pRAF-1 S259 levels remained unaltered in the other cell lines. Inhibition of PI3K/AKT signalling by Ly294002 relieves the AKT mediated phosphorylation of RAF-1, thereby restoring MAPK signalling. CONCLUSIONS: Here we show that hybrid cells could evolve exhibiting a differential active RAF-AKT crosstalk. Because PI3K/AKT signalling has been chosen as a target for anti-cancer therapies our data might point to a possible severe side effect of AKT targeted cancer therapies. Inhibition of PI3K/AKT signalling in RAF-AKT crosstalk positive cancer (hybrid) cells could result in a progression of these cells. Thus, not only the receptor (activation) status, but also the activation of signal transduction molecules should be analysed thoroughly prior to therapy.