Effects of extracellular matrix protein functionalized fluid membrane on cell adhesion and matrix remodeling
Effects of extracellular matrix protein functionalized fluid membrane on cell adhesion and matrix remodeling
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DOI：
10.1016/j.biomaterials.2010.05.076
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发表时间：
2010-09-01
期刊：
影响因子：
14
通讯作者：
Chang，Ying-Chih
中科院分区：
文献类型：
--
作者：
Huang，Chun-Jen;Tseng，Po-Yuan;Chang，Ying-Chih
为了研究生物材料涂层介导的细胞反应和细胞外基质蛋白重塑，我们提出了一种含有蛋白质共轭支撑的脂质双层(SLB)的仿生构建物作为细胞培养平台。通过将I型胶原和/或纤维连接蛋白连接到N-羟基磺基琥珀酰亚胺功能化的SLB上，制备了单组分或多组分的蛋白质结合的SLB。用耗散石英晶体微天平对蛋白质共轭体系进行了定量表征。将NIH3T3成纤维细胞在模型构建物和氧等离子体处理的聚苯乙烯(PSO)上进行培养，以进行平行比较。通过光漂白后的荧光恢复来评估蛋白质偶联和细胞培养后SLB的迁移率的延迟。对所得的细胞形态、吸附动力学和体细胞动力学进行了显微镜观察。我们发现，在基于SLB的培养中，3T3成纤维细胞在含有纤维连接蛋白的表面铺展大小和细胞数最大。然而，在涂有蛋白质的PSO表面上，没有观察到所有蛋白质含量的明显差异。免疫荧光染色结果显示，3T3细胞在PSO表面对内源性纤维连接蛋白的吸附显著高于SLB表面。这表明SLB的防污染特性在阻止3T3细胞有效重塑其微环境方面发挥了重要作用，而细胞可以很容易地重塑非特异性吸附倾向表面，如基于PSO的平台。总之，蛋白质结合的SLB表面可以作为一个平台，用于确定和调节细胞特异性结合以及随后与细胞外环境发生的信号事件。
In order to study cellular responses and extracellular matrix protein remodeling mediated by biomaterials coating, we proposed a biomimetic construct containing protein-conjugated supported lipid bilayers (SLBs) as a cell culture platform. Single or multi-component proteins-bound SLBs were fabricated by conjugating type I collagen and/or fibronectin on the N-hydroxysulfosuccinimide-functionalized SLBs. The proposed protein-conjugated systems were quantitatively characterized by the quartz crystal microbalance with dissipation. NIH 3T3 fibroblasts were cultured on the model constructs and on oxygen plasma pretreated polystyrene (PSo) for parallel comparison. The retards of mobility of SLB after protein conjugation and cell culture were estimated by fluorescence recovery after photobleaching. The resulting cell morphology, adsorption kinetics and somatic dynamics were examined microscopically. We found that, on the SLB based cultures, the largest spreading size and cell number counts of 3T3 fibroblasts were found on the fibronectin containing surfaces. However, on the protein-coated PSo surfaces, no such distinguishable differences can be observed on all protein contents. Immunofluorescent staining results revealed that adsorption of endogenously produced fibronectin by 3T3 cells on PSo based surfaces is significantly more than that on SLB based surfaces. This suggests that the anti-fouling nature of underneath SLBs have played an important role in preventing 3T3 cells from effectively remodeling their microenvironment, whereas cells can easily remodel the nonspecific adsorption prone surfaces such as PSo based platforms. In summary, the protein conjugated SLB surfaces can serve as a platform for determining and regulating cell specific binding and subsequent signaling events with extracellular environments.