Ivanova Svetlana I, Chakarov Stoyan, Momchilova Albena, Pankov Roumen
Department of Cytology, Histology and Embryology, Faculty of Biology, Sofia University "St. Kliment Ohridski", Sofia, Bulgaria.
Department of Biochemistry, Faculty of Biology, Sofia University "St. Kliment Ohridski", Sofia, Bulgaria.
Mater Sci Eng C Mater Biol Appl. 2017 Sep 1;78:230-238. doi: 10.1016/j.msec.2017.04.071. Epub 2017 Apr 17.
The present study describes a live-cell biosensor, suitable for general evaluation of adhesion qualities of different substrates. It is based on NIH/3T3 fibroblast cell line stably expressing fusion fluorescently tagged proteins mCherry-vinculin and GFP-tensin as quantifiable markers for assessment not only of focal but also of fibrillar contacts. Four measurable parameters - spreading, polarization and development of focal and fibrillar adhesions were used to standardize the adhesion of biosensor cells after plating on five substrates of natural origin - fibronectin, vitronectin, laminin-111, laminin-521 and collagen type I. The obtained set of values (adhesion quality map) were utilized to describe the default biosensor behavior and as a standard for evaluation of surface biocompatibility of materials with unknown adhesive properties. To demonstrate the applicability of the biosensor we studied two PDMS-based artificial materials. The results demonstrated the superior adhesive properties of the poly(acrylic acid)-containing polymer (PDMS-PAA) over that of poly(vinyl pyrrolidone) copolymer (PDMS-PVP), and pointed out the formation of focal adhesions as a parameter for possible further improvements.
本研究描述了一种活细胞生物传感器，适用于对不同底物的粘附特性进行综合评估。它基于稳定表达融合荧光标记蛋白mCherry-纽蛋白和GFP-张力蛋白的NIH/3T3成纤维细胞系，作为不仅用于评估粘着斑而且用于评估纤维状接触的可量化标记。使用四个可测量参数——铺展、极化以及粘着斑和纤维状粘着的形成，来规范生物传感器细胞接种在五种天然来源底物（纤连蛋白、玻连蛋白、层粘连蛋白-111、层粘连蛋白-521和I型胶原）上后的粘附情况。所获得的一组值（粘附质量图）被用于描述生物传感器的默认行为，并作为评估具有未知粘附特性材料的表面生物相容性的标准。为了证明该生物传感器的适用性，我们研究了两种基于聚二甲基硅氧烷（PDMS）的人工材料。结果表明，含聚丙烯酸的聚合物（PDMS-PAA）比聚乙烯吡咯烷酮共聚物（PDMS-PVP）具有更优异的粘附特性，并指出粘着斑的形成是可能进一步改进的一个参数。
