Abstract
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.
Highlights
A plethora of biomaterials used as scaffolds for tissue engineering as well as their influence on the quality of the generated tissue according to their specific properties have been described previously
The spindle-shaped and asymmetric morphology of a single fibroblast in the more detailed SEM revealed a polarity with a more and a less convex side of the fibroblast, which is defined as part of cell migration processes (Fig. 2b) [29]
These findings could be confirmed by analyzing the assembly of the actin filament bundles, which could be regarded as intracellular lines of force (Fig. 3): Concentration of the actin cortex inside the lamellopodium was observed, which counts as characteristic for migratory processes (Fig. 3, cell on the right)
Summary
A plethora of biomaterials used as scaffolds for tissue engineering as well as their influence on the quality of the generated tissue according to their specific properties have been described previously. It has been discussed that, foremost, an ideal tissue-engineering scaffold should act as replacement for the tissue that should be restored and have comparable mechanical attributes [1] It should bridge the gap, carry histologically typical cells and guide tissue repair. Research has focused on the use of degradable scaffold materials, especially synthetic polymers like polyglycolic acid (PGA) or polylactic acid (PLA) [2]. While these often promise very good moldability, they often have poor mechanical properties. As untreated silkworm silk has been found to be quite immunogenic, an alternative could be spider silk Does it own unique mechanical properties, its biocompatibility has been shown already in vivo. We used native spider dragline silk which is known as the strongest fibre in nature
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