Influence of CO 2 on the micro-structural properties of spider dragline silk: X-ray microdiffraction results

Rößle, Manfred; Sapede, Daniel; Vollrath, Fritz

doi:10.1007/s00114-003-0482-8

Cited by 20 publications

(25 citation statements)

References 20 publications

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“…Briefly, the spiders were atraumatically fixed and immobilized on styrophor cubes with gauze and needles, and the major ampullate gland was stimulated by pulling the dragline out of the anterior spinneret mechanically. Spiders were not harmed during the harvesting process and no anaesthesia was used to avoid pH changes induced by carbon dioxide (CO 2 ) anaesthesia [53].…”

Section: Methodsmentioning

confidence: 99%

Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

et al. 2010

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BackgroundSeveral materials have been used for tissue engineering purposes, since the ideal matrix depends on the desired tissue. Silk biomaterials have come to focus due to their great mechanical properties. As untreated silkworm silk has been found to be quite immunogenic, an alternative could be spider silk. Not only does it own unique mechanical properties, its biocompatibility has been shown already in vivo. In our study, we used native spider dragline silk which is known as the strongest fibre in nature.Methodology/Principal FindingsSteel frames were originally designed and manufactured and woven with spider silk, harvesting dragline silk directly out of the animal. After sterilization, scaffolds were seeded with fibroblasts to analyse cell proliferation and adhesion. Analysis of cell morphology and actin filament alignment clearly revealed adherence. Proliferation was measured by cell count as well as determination of relative fluorescence each after 1, 2, 3, and 5 days. Cell counts for native spider silk were also compared with those for trypsin-digested spider silk. Spider silk specimens displayed less proliferation than collagen- and fibronectin-coated cover slips, enzymatic treatment reduced adhesion and proliferation rates tendentially though not significantly. Nevertheless, proliferation could be proven with high significance (p<0.01).Conclusion/SignificanceNative spider silk does not require any modification to its application as a biomaterial that can rival any artificial material in terms of cell growth promoting properties. We could show adhesion mechanics on intracellular level. Additionally, proliferation kinetics were higher than in enzymatically digested controls, indicating that spider silk does not require modification. Recent findings concerning reduction of cell proliferation after exposure could not be met. As biotechnological production of the hierarchical composition of native spider silk fibres is still a challenge, our study has a pioneer role in researching cellular mechanics on native spider silk fibres.

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Section: Methodsmentioning

confidence: 99%

Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

et al. 2010

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“…To obtain single fiber major ampullate dragline silk, the spider was anesthetized using carbon dioxide and subsequently immobilized on a polystyrene block with its underside facing upward. The spider was left to regain consciousness for 30 min to eliminate any effect of the carbon dioxide on the spinning process (Riekel et al, 2004). Under an optical stereo microscope (Leica MZ6), a single major ampullate dragline thread was separated from other silks and taken up onto the reeling device.…”

Section: Collection Of Single Fiber Dragline Silkmentioning

confidence: 99%

Revealing Spider Silk's 3D Nanostructure Through Low Temperature Plasma Etching and Advanced Low-Voltage SEM

et al. 2019

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The excellent mechanical properties of spider dragline silk are closely linked to its multiscale hierarchical structuring which develops as it is spun. If this is to be understood and mimicked, multiscale models must emerge which effectively bridge the length scales. This study aims to contribute to this goal by exposing structures within Nephila dragline silk using low-temperature plasma etching and advanced Low Voltage Scanning Electron Microscopy (LV-SEM). It is shown that Secondary Electron Hyperspectral Imaging (SEHI) is sensitive to compositional differences on both the micro and nano scale. On larger scales it can distinguish the lipids outermost layer from the protein core, while at smaller scales SEHI is effective in better resolving nanostructures present in the matrix. Key results suggest that the silks spun at lower reeling speeds tend to have a greater proportion of smaller nanostructures in closer proximity to one-another in the fiber, which we associate with the fiber's higher toughness but lower stiffness. The bimodal size distribution of ordered domains, their radial distribution, nanoscale spacings, and crucially their interactions may be key in bridging the length scale gaps which remain in current spider silk structure-property models. Ultimately this will allow successful biomimetic implementation of new models.

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“…Diffraction patterns of carbon research papers fibres collected using an X-ray nanobeam and a two-dimensional detector have been used to identify the buckling of bent fibres under compression and to compare the strain and compression behaviour of fibres from different manufacturing processes (Loidl et al, 2005). The orientation of macromolecules as determined by wide-angle X-ray scattering has been used to demonstrate that increased exposure of spiders to carbon dioxide alters the amino acid makeup of their extruded silk, consequently altering its material properties (Riekel et al, 2004). Crystal orientation measurements have revealed a preferred orientation in tooth enamel which varies relative to the biting surface and to the position on the tooth, as well as variations in lattice parameters travelling from the outer enamel to the dentine junction, hypothesized to result from variations in chemical composition (Al-Jawad et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Full-section otolith microtexture imaged by local-probe X-ray diffraction

et al. 2018

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An optimized synchrotron-based X-ray diffraction method is described for the direct and efficient measurement of crystallite phase and orientation at micrometre resolution across textured polycrystalline samples of millimetre size (high scale dynamics) within a reasonable time frame. The method is demonstrated by application to biomineral fish otoliths. Otoliths are calcium carbonate accretions formed in the inner ears of vertebrates. Fish otoliths are essential biological archives, providing information for individual age estimation, the study of population dynamics and fish stock management, as well as past environmental and climatic conditions from archaeological specimens.Here, X-ray diffraction mapping is discussed as a means of describing the mineralogical structure and microtexture of otoliths. Texture maps could be generated with a few a priori hypotheses on the aragonitic system. Full-section imaging allows quantitative intercomparison of crystal orientation coupled to microstructural description, across the zones of the otoliths that represent distinctive mineral organization. It reveals the extents of these regions and their internal textural structure. Characterization of structural and textural correlations across whole images is therefore proposed as a complementary approach to investigate and validate the local in-depth nanometre-scale study of biominerals. The estimation of crystallite size and orientational distribution points to diffracting domains intermediate in size between the otolith nanogranules and the crystalline units, in agreement with recently reported results.

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Influence of CO 2 on the micro-structural properties of spider dragline silk: X-ray microdiffraction results

Cited by 20 publications

References 20 publications

Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

Revealing Spider Silk's 3D Nanostructure Through Low Temperature Plasma Etching and Advanced Low-Voltage SEM

Full-section otolith microtexture imaged by local-probe X-ray diffraction

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