Oliveira Catarina, Costa-Pinto Ana R, Reis Rui L, Martins Albino, Neves Nuno M
3B's Research Group - Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine , AvePark, Zona Industrial da Gandra S. Cláudio do Barco, 4806-909 Caldas das Taipas, Guimarães, Portugal.
Biomacromolecules. 2014 Jun 9;15(6):2196-205. doi: 10.1021/bm500346s. Epub 2014 May 22.
The immobilization of biomolecules at the surface of different biomedical devices has attracted enormous interest in order to enhance their biological functionality at the cellular level. This work aims to develop a biofunctional polymeric substrate capable of selectively binding growth factors (GFs) of interest from a pool of proteins present in a biological fluid: platelet lysate (PL). To achieve this goal, the surface of electrospun PCL nanofibers needs to be activated and functionalized to be able to insert chemical groups for the immobilization of antibodies. After determining the maximum immobilization capacity of each antibody, TGF-β1 (12 μg mL(-1)), bFGF (8 μg mL(-1)), and VEGF (4 μg mL(-1)), the next step was to confirm their bioavailability using recombinant proteins. The binding efficiency of PL-derived GFs was of 84-87% for TGF-β1, 55-64% for bFGF, and 50-59% for VEGF. Cellular assays confirmed the biological activity of the bound VEGF (both recombinant and PL-derived). Multiple antibodies (i.e., bFGF and VEGF) were also immobilized over the same structure in a mixed or side-by-side fashion. Using both autologous biological fluids and cells, it is possible to use this platform to implement very effective and personalized therapies that can be tailored to specific medical conditions.
为了在细胞水平上增强生物分子的生物学功能,将其固定在不同生物医学装置的表面已引起了极大的关注。这项工作旨在开发一种生物功能聚合物基质,该基质能够从生物流体——血小板裂解液(PL)中存在的一组蛋白质中选择性地结合感兴趣的生长因子(GFs)。为实现这一目标,需要对电纺聚己内酯(PCL)纳米纤维的表面进行活化和功能化,以便能够引入用于固定抗体的化学基团。在确定每种抗体(转化生长因子-β1(TGF-β1,12μg/mL)、碱性成纤维细胞生长因子(bFGF,8μg/mL)和血管内皮生长因子(VEGF,4μg/mL))的最大固定容量后,下一步是使用重组蛋白确认它们的生物利用度。PL衍生的GFs对TGF-β1的结合效率为84%-87%,对bFGF为55%-64%,对VEGF为50%-59%。细胞试验证实了结合的VEGF(重组的和PL衍生的)的生物活性。多种抗体(即bFGF和VEGF)也以混合或并排的方式固定在同一结构上。使用自体生物流体和细胞,可以利用这个平台实施非常有效且个性化的治疗方法,这些方法可以针对特定的医疗状况进行定制。