title: Synergistic protein secretion by mesenchymal stromal cells seeded in 3D scaffolds and circulating leukocytes in physiological flow.
abstract: Mesenchymal stromal cells (MSC) play an important role in natural wound healing via paracrine and juxtacrine signaling to immune cells. The aim of this study was to identify the signaling factors secreted by preseeded cells in a biomaterial and their interaction with circulating leukocytes, in the presence of physiological biomechanical stimuli exerted by the hemodynamic environment (i.e. strain and shear flow). Electrospun poly(ε-caprolactone)-based scaffolds were seeded with human peripheral blood mononuclear cells (PBMC) or MSC. Protein secretion was analyzed under static conditions and cyclic strain. Subsequently, the cross-talk between preseeded cells and circulating leukocytes was addressed by exposing the scaffolds to a suspension of PBMC in static transwells and in pulsatile flow. Our results revealed that PBMC exposed to the scaffold consistently secreted a cocktail of immunomodulatory proteins under all conditions tested. Preseeded MSC, on the other hand, secreted the trophic factors MCP-1, VEGF and bFGF. Furthermore, we observed a synergistic upregulation of CXCL12 gene expression and a synergistic increase in bFGF protein production by preseeded MSC exposed to PBMC in pulsatile flow. These findings identify CXCL12 and bFGF as valuable targets for the development of safe and effective acellular instructive grafts for application in in situ cardiovascular regenerative therapies.
authors.affiliation: Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: v.ballotta@tue.nl.
authors.forename: Virginia
authors.initials: V
authors.lastname: Ballotta
authors.affiliation: Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: a.i.p.m.smits@tue.nl.
authors.forename: Anthal I P M
authors.initials: AI
authors.lastname: Smits
authors.affiliation: Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. Electronic address: a.driessen@tue.nl.
authors.forename: Anita
authors.initials: A
authors.lastname: Driessen-Mol
authors.affiliation: Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands; Institute for Complex Molecular Systems (ICMS), Eindhoven, The Netherlands. Electronic address: c.v.c.bouten@tue.nl.
authors.forename: Carlijn V C
authors.initials: CV
authors.lastname: Bouten
authors.affiliation: Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands; Institute for Complex Molecular Systems (ICMS), Eindhoven, The Netherlands. Electronic address: f.p.t.baaijens@tue.nl.
authors.forename: Frank P T
authors.initials: FP
authors.lastname: Baaijens
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000378:metabolism*; D054377:Chemokine CXCL12 / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D011091:Polyesters / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D014945:Wound Healing / Q000502:physiology
chemical_list: C111835:CCL2 protein, human; C515255:CXCL12 protein, human; D018932:Chemokine CCL2; D054377:Chemokine CXCL12; D011091:Polyesters; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; C016240:polycaprolactone
keywords: Cell signaling; Chemotaxis; Co-culture; Fibroblast growth factor; Monocytes; Shear stress