Sharma Vaibhav, Blackwood Keith A, Haddow David, Hook Lilian, Mason Chris, Dye Julian F, García-Gareta Elena
RAFT Institute of Plastic Surgery, Mount Vernon Hospital, Northwood, HA6 2RN, UK.
Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT, UK.
Biochim Open. 2015 Oct 24;1:40-50. doi: 10.1016/j.biopen.2015.10.001. eCollection 2015.
Composite biomaterials made from synthetic and protein-based polymers are extensively researched in tissue engineering. To successfully fabricate a protein-polymer composite, it is critical to understand how strongly the protein binds to the synthetic polymer, which occurs through protein adsorption. Currently, there is no cost-effective and simple method for characterizing this interfacial binding. To characterize this interfacial binding, we introduce a simple three-step method that involves: 1) synthetic polymer surface characterisation, 2) a quick, inexpensive and robust novel immuno-based assay that uses protein extraction compounds to characterize protein binding strength followed by 3) an in vitro 2D model of cell culture to confirm the results of the immuno-based assay. Fibrinogen, precursor of fibrin, was adsorbed (test protein) on three different polymeric surfaces: silicone, poly(acrylic acid)-coated silicone and poly(allylamine)-coated silicone. Polystyrene surface was used as a reference. Characterisation of the different surfaces revealed different chemistry and roughness. The novel immuno-based assay showed significantly stronger binding of fibrinogen to both poly(acrylic acid) and poly(allylamine) coated silicone. Finally, cell studies showed that the strength of the interaction between the protein and the polymer had an effect on cell growth. This novel immuno-based assay is a valuable tool in developing composite biomaterials of synthetic and protein-based polymers with the potential to be applied in other fields of research where protein adsorption onto surfaces plays an important role.
由合成聚合物和蛋白质基聚合物制成的复合生物材料在组织工程领域得到了广泛研究。要成功制备蛋白质-聚合物复合材料,关键在于了解蛋白质与合成聚合物的结合强度,这种结合是通过蛋白质吸附实现的。目前,尚无一种经济高效且简便的方法来表征这种界面结合。为了表征这种界面结合,我们引入了一种简单的三步法,该方法包括:1)合成聚合物表面表征;2)一种快速、廉价且可靠的基于免疫的新型检测方法,该方法使用蛋白质提取化合物来表征蛋白质结合强度,随后是3)细胞培养的体外二维模型,以确认基于免疫的检测结果。纤维蛋白原(纤维蛋白的前体)被吸附(测试蛋白)在三种不同的聚合物表面上:硅酮、聚(丙烯酸)涂层硅酮和聚(烯丙胺)涂层硅酮。聚苯乙烯表面用作参考。对不同表面的表征揭示了不同的化学性质和粗糙度。基于免疫的新型检测方法表明,纤维蛋白原与聚(丙烯酸)和聚(烯丙胺)涂层硅酮的结合明显更强。最后,细胞研究表明,蛋白质与聚合物之间相互作用的强度对细胞生长有影响。这种基于免疫的新型检测方法是开发合成聚合物和蛋白质基聚合物复合生物材料的宝贵工具,有望应用于蛋白质吸附到表面起重要作用的其他研究领域。
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