Department of Plastic, Hand and Reconstructive Surgery, Hannover Medical School, Hannover, Germany.
PLoS One. 2010 Aug 9;5(8):e12032. doi: 10.1371/journal.pone.0012032.
Several 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 FINDINGS: Steel 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/SIGNIFICANCE: Native 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.
由于理想的基质取决于所需的组织，因此已经有几种材料被用于组织工程目的。由于其出色的机械性能，丝生物材料成为研究热点。由于未经处理的家蚕丝被发现具有很强的免疫原性，因此另一种选择可能是蜘蛛丝。它不仅具有独特的机械性能，而且其生物相容性已经在体内得到证实。在我们的研究中，我们使用了天然蜘蛛牵引丝，它被认为是自然界中最强的纤维。
方法/主要发现：最初设计并制造了钢框架，并使用蜘蛛丝进行编织，直接从动物身上收获牵引丝。经过消毒后，支架被接种成纤维细胞，以分析细胞增殖和黏附。细胞形态和肌动蛋白丝排列的分析清楚地揭示了黏附。通过细胞计数以及第 1、2、3 和 5 天后相对荧光的测定来测量增殖。还将天然蜘蛛丝的细胞计数与经胰蛋白酶消化的蜘蛛丝的细胞计数进行了比较。与胶原蛋白和纤维连接蛋白涂层载玻片相比，蜘蛛丝标本的增殖较少，尽管酶处理并未显著降低黏附率和增殖率，但仍能显著证明增殖（p<0.01）。
结论/意义：天然蜘蛛丝在应用时无需任何修饰即可作为生物材料，在促进细胞生长方面可与任何人工材料相媲美。我们可以在细胞内水平上展示黏附力学。此外，增殖动力学高于酶消化对照，表明蜘蛛丝不需要修饰。最近关于暴露后细胞增殖减少的发现无法得到证实。由于天然蜘蛛丝纤维的层次结构的生物制造仍然是一个挑战，因此我们的研究在研究天然蜘蛛丝纤维的细胞力学方面具有先驱作用。
