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Purpose:
Patients with primary open-angle glaucoma (POAG) show a stiffer peripapillary sclera, reactive astrocytes and a remodeled lamina cribrosa (LC). The changes are thought to be mediated by TGFβ2 and its downstream mediator CTGF. Recently we developed a murine glaucoma model by overexpressing CTGF in the anterior eye (βb1CTGF). In this study we investigated the glial lamina region of βb1CTGF mice, and the changes of astrocytes in response to CTGF and TGFβ2 as well as increasing substratum stiffness.
Methods:
Tangential sections of the glial LC of 2-month-old βb1CTGF mice and their wild-type littermates (WT) were stained with phalloidin and antibodies against GFAP, CTGF and fibronectin (FN). Murine optic nerve (ON) astrocytes from CD1 mice were isolated, cultured and characterized by GFAP staining. The astrocytes were treated with TGFβ2 (1ng/ml) and CTGF (50ng/ml and 100ng/ml). In addition, the cells were seeded on PDMS substrata with different stiffness (10, 30 and 60 kPa). Treated cells were analyzed by Western blotting, real-time RT-PCR and immunohistochemistry. Wound healing assays were performed to analyze migration rate following growth factor treatment.
Results:
βb1CTGF mice showed a massive increase in CTGF and GFAP in the glial LC when compared with WT mice. Moreover, an increase in FN staining and phalloidin-labeled actin was observed in the peripapillary sclera. Murine ON astrocytes reacted on increased substrate stiffness by increasing the synthesis of GFAP, vimentin and CTGF. Treatment of the cells with TGFβ2 and CTGF led to an enhanced migration rate. In addition, treatment resulted in an increased expression and synthesis of ECM proteins, including FN, tropoelastin, collagen type I and III. The in vitro findings correlated with those seen in the glaucoma mouse model.
Conclusions:
We conclude that changes in the ECM of LC and peripapillary sclera alter their biomechanical properties and thereby induce reactive changes in resident astrocytes. The reactive changes induced by higher stiffness of their surrounding ECM give rise to a self-amplifying process that includes increased TGFβ2/CTGF signaling and leads to synthesis of ECM and cytoskeletal proteins, a process that in turn augments the stiffness at the optic nerve head (ONH). Such a scenario may finally result in a vicious circle as the causative mechanism for ONH deformation in POAG.
Hysteresis in dynamic modulus, loss factor and normal forces of magnetoactive elastomers (MAEs) comprising various proportions of small (3-5 mu m) and large (50-60 mu m) ferromagnetic particles are experimentally studied using dynamic torsion performed at a fixed oscillation frequency in varying DC magnetic fields. It is shown that hysteresis is a characteristic feature of MAEs observed both under increasing/decreasing magnetic field strength and increasing/decreasing strain amplitude. This hysteresis is attributed to the specific rearrangement of the magnetic filler network under simultaneously applied magnetic field and shear deformation. Rheological properties of the magnetic filler network formed in the magnetic field and, therefore, the rheological properties of MAEs depend strongly on the filler composition and the magnetic field magnitude. Larger magnetic particles and higher magnetic fields provide stronger magnetic networks. Both factors result in the extension of the linear viscoelastic regime to larger strain amplitudes and lead to higher values of shear storage and loss moduli. It is found that the hysteresis width maximises at an intermediate magnetic field where it is attributed to the balance between elastic and magnetic particle interactions. This is apparently where the most significant restructuring of the magnetic network occurs. The hysteresis width decreases with increasing fraction of large particles in the magnetic filler. The loss factor grows significantly when the magnetic network is physically broken by large strains gamma > 1%. A huge (more than one order of magnitude) increase of normal force at maximum magnetic field strengths is observed. It is predicted that any physical quantity depending on the internal structuring of the magnetic filler should demonstrate hysteresis either with a changing magnetic field and constant deformation amplitude or under variable deformation in a constant magnetic field. (C) 2015 Elsevier Ltd. All rights reserved.
During recent years, there has been a trend to replace traditional lab courses by project-based learning activities. However, there is currently little scientific evidence for educational improvements through such changes. Some years ago, a lab course on electrical measurements in our bachelor Electrical Engineering and Information Technology was modified from single experiments to projects. This change led to a significant increase in the necessary manpower and lab costs. To justify these higher efforts, it is important to investigate if the learning success increased. We carried out the surveys and collected the statistical data from the students who attended this course. In order to see if the students appreciated this lab course and if there was an objective increase of the students' knowledge, these results were evaluated. Evaluation of the grades in the lecture and lab courses shows that the increase of the students' theoretical and practical knowledge is statistically significant.