Cytoskeleton systems contribute differently to the functional intrinsic properties of chondrospheres.

Cytoskeleton systems, actin microfilaments, microtubules (MTs) and intermediate filaments (IFs) provide the biomechanical stability and spatial organization in cells. To understand the specific contributions of each cytoskeleton systems to intrinsic properties of spheroids, we've scrutinized the effects of the cytoskeleton perturbants, cytochalasin D (Cyto D), nocodazole (Noc) and withaferin A (WFA) on fusion, spreading on adhesive surface, morphology and biomechanics of chondrospheres (CSs). We confirmed that treatment with Cyto D but not with Noc or WFA severely affected CSs fusion and spreading dynamics and significantly reduced biomechanical properties of cell aggregates. Noc treatment affected spheroids spreading but not the fusion and surprisingly enhanced their stiffness. Vimentin intermediate filaments (VIFs) reorganization affected CSs spreading only. The analysis of all three cytoskeleton systems contribution to spheroids intrinsic properties was performed for the first time.

Genotoxic effect and rat hepatocyte death occurred after oxidative stress induction and antioxidant gene downregulation caused by long term fluoride exposure.

Studies focusing on possible genotoxic effects of excess fluoride are contradictory and inconclusive. Currently, studies have reported a probable link to oxidative stress, DNA damage and apoptosis induced by fluoride in rat hepatocytes. We developed an in vivo study administering three doses of fluoride by gavage given to rats for 60 day. Micronucleus test was applied to investigate genotoxic potential of fluoride. The TUNEL method determined DNA fragmentation and apoptosis. Biochemical parameters to investigate mitochondrial swelling and oxidative stress. Semi-quantitative RT-PCR and immunostaining to determine mRNA and protein expression of antioxidant enzymes. Analyses of the hepatic function and morphology were performed. Our results revealed the genotoxic potential of fluoride but did not confirm mitochondrial swelling nor an increase of positive TUNEL labelling induced by fluoride, indicating absence of apoptosis. Oxidative stress induction was confirmed and is probably associated to DNA damage. Cell death events such as empty nuclear spaces, cytoplasm degeneration, nuclear pyknosis, karyorrhexis and karyorrhexis followed by karyolysis were observed. Hepatic function did not appear to be significantly modified makes no evidence of necrosis and suggesting other cell death pathway, the autophagic. In conclusion, prolonged fluoride intake at chosen concentrations caused imbalance of the cellular oxidative state, affected DNA and disrupted cellular homeostasis. It is recommended that fluoride supplementation requires a fresh consideration in light of the current study.

[SPREADING OF TISSUE SPHEROIDS FROM PRIMARY HUMAN FIBROBLASTS ON THE SURFACE OF MICROFIBROUS ELECTROSPUN POLYURETHANE MATRIX (A scanning electron microscopic study)].

Tissue spheroids biofabricated from primary human fibroblasts using non-adhesive agarose forms, were placed by 3D bioprinter on the surface of microfibrous electrospun matrix. It was demonstrated that tissue spheroids attached to the surface of matrix during several hours and then gradually spread for several days which indicates high level of biocompatibiity of electrospun microfibrous polyurethane matrix. During this activity, human fibroblasts used processes of leading cell borders for initial step of attachment to matrix filaments. Tissue constructions formed during spreading of tissue spheroids on the surface of electrospun microfibrous polyurethane matrix seem to be a perspective technology platform for development of new methods of biofabrication and 3D bioprinting.

Walking economy of adults with Down syndrome.

This study intended to investigate walking economy (WE) in response to different treadmill speeds and grades in adults with Down syndrome (DS) and in non-disabled controls. Eighteen participants (14 males; 4 females) with DS (33.6+/-7.6 years) and 16 non-disabled (12 males, 4 females) controls (33.3+/-8.0 years) performed submaximal (2.5 km . h (-1) and 4 km . h (-1) at 0% grade; 4 km . h (-1) at 2.5% and 5% grade, for 5 min each) and maximal treadmill tests with metabolic and heart rate measurements. Oxygen uptake (VO(2)) was not different between groups at rest or during the slowest treadmill speed. However, at faster speeds and increased grades, adults with DS presented lower WE than controls (p<0.0001). Subsequent analyses revealed that, despite showing higher delta VO(2) response to the selected speed increments (p<0.0001), individuals with DS produced similar VO(2) increase as controls to grade variations. Therefore, adults with DS exhibit lower WE than non-disabled controls at a speed faster than their preferred walking speed. Additionally, in comparison to controls, individuals with DS show a greater change in energy expenditure with a change in walking speed. In conclusion, lower WE in individuals with DS is mainly related to their inability to adapt efficiently to positive variations in walking speed.