title: Biofunctional nanofibrous substrate comprising immobilized antibodies and selective binding of autologous growth factors.
abstract: 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.
authors.affiliation: 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.
authors.forename: Catarina
authors.initials: C
authors.lastname: Oliveira
authors.forename: Ana R
authors.initials: AR
authors.lastname: Costa-Pinto
authors.forename: Rui L
authors.initials: RL
authors.lastname: Reis
authors.forename: Albino
authors.initials: A
authors.lastname: Martins
authors.forename: Nuno M
authors.initials: NM
authors.lastname: Neves
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D005346:Fibroblast Growth Factors / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D055767:Immobilized Proteins / Q000032:analysis / Q000378:metabolism*; D051379:Mice; D057139:Nanofibers / Q000737:chemistry; D011485:Protein Binding / Q000502:physiology; D013379:Substrate Specificity / Q000502:physiology; D053773:Transforming Growth Factor beta1 / Q000032:analysis / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis / Q000378:metabolism*
chemical_list: D000906:Antibodies; D055767:Immobilized Proteins; D053773:Transforming Growth Factor beta1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D005346:Fibroblast Growth Factors