National Nanotechnology Laboratory, Institute Nanoscience CNR, NNL, CNR-NANO, via Arnesano, Lecce, Italy.
Integr Biol (Camb). 2013 Aug;5(8):1057-66. doi: 10.1039/c3ib40064f.
We have recently reported initial results concerning an original approach to introduce additional properties into fibrillar proteins produced by live fibroblasts and extruded into the ECM. The key to such an approach was biocompatible, fluorescent and semiconducting synthetic molecules which penetrated spontaneously the cells and were progressively encompassed via non-bonding interactions during the self-assembly process of the proteins, without altering cell viability and reproducibility. In this paper we demonstrate that the intracellular secretion of fluorescent microfibers can be generalized to living primary and immortalized human/mouse fibroblasts. By means of real-time single-cell confocal microscopy we show that the fluorescent microfibers, most of which display helical morphology, are generated by intracellular coding of the synthetic molecules. We also describe co-localization experiments on the fluorescent microfibers isolated from the cell milieu demonstrating that they are mainly made of type-I collagen. Finally, we report experimental data indicating that the embedded synthetic molecules cause the proteins not only to be fluorescent but also capable of electrical conductivity.
我们最近报道了一种将额外性质引入由活成纤维细胞产生并挤出到细胞外基质中的纤维状蛋白质的原始方法的初步结果。这种方法的关键是生物相容性、荧光和半导体合成分子，它们能够自发穿透细胞，并在蛋白质的自组装过程中通过非键相互作用逐渐被包含，而不改变细胞活力和重现性。在本文中，我们证明了荧光微纤维的细胞内分泌可以推广到活的原代和永生化的人/鼠成纤维细胞。通过实时单细胞共焦显微镜，我们表明荧光微纤维是通过合成分子的细胞内编码产生的，其中大多数呈现螺旋形态。我们还描述了从细胞环境中分离出的荧光微纤维的共定位实验，证明它们主要由 I 型胶原蛋白组成。最后，我们报告了实验数据，表明嵌入的合成分子不仅使蛋白质具有荧光性，而且还具有导电性。
