The development of new capillary networks in engineered constructs is essential for their survival and their integration with the host tissue. It has recently been demonstrated that ELR-based hydrogels encoding different bioactivities are able to modulate their interaction with the host after injection or implantation, as indicated by an increase in cell adhesion and the ability to trigger vascularization processes. Accordingly, the aim of this study was to increase their angiogenic ability both in vitro and in vivo using a small VEGF mimetic peptide named QK, which was tethered chemically to ELR-based hydrogels containing cell-adhesion sequences in their backbone, such as REDV and RGD, as well as a proteolytic site (VGVAPG). In vitro studies were performed using a co-culture of endothelial and fibroblast cells encapsulated into the ELR-based hydrogels in order to determine cell proliferation after 21 days of culture, as well as the number of cell-cell interactions. It was found that although the presence of this peptide does not influence the morphological and rheological properties of these hydrogels, it has an effect on cell behaviour, inducing an increase in cell proliferation and the formation of endothelial cell clusters. In vivo studies demonstrate that the QK peptide enhances the formation of prominent functional capillaries at three weeks post-injection, as confirmed by H&E staining and CD31 immunohistochemistry. The newly formed functional microvasculature ensures perfusion and connection with surrounding tissues. These results show that ELR-QK hydrogels increase capillary network formation and are therefore attractive candidates for application in tissue regeneration, for example for the treatment of cardiovascular diseases such as myocardial infarction or ischemia.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Jenkins DD, Yang GP, Lorenz HP, Longaker MT, Sylvester KG. Tissue engineering and regenerative medicine. Clin Plast Surg. 2003;30:581–8.
Place ES, Evans ND, Stevens MM. Complexity in biomaterials for tissue engineering. Nat Mater. 2009;8:457.
Dhandayuthapani B, Yoshida Y, Maekawa T, Kumar DS. Polymeric scaffolds in tissue engineering application: a review. Int J Polym Sci. 2011;2011:3–7, 10–11.
Lovett M, Lee K, Edwards A, Kaplan DL. Vascularization strategies for tissue engineering. Tissue Eng Part B Rev. 2009;15:353–70.
West JL, Moon JJ. Vascularization of engineered tissues: approaches to promote angiogenesis in biomaterials. Curr Top Med Chem. 2008;8:300–10.
Javier Arias F, Santos M, Fernández-Colino A, Pinedo G, Girotti A. Recent contributions of elastin-like recombinamers to biomedicine and nanotechnology. Curr Top Med Chem. 2014;14:819–36.
MacEwan SR, Chilkoti A. Elastin‐like polypeptides: biomedical applications of tunable biopolymers. Biopolymers. 2010;94:60–77.
Girotti A, Fernández‐Colino A, López IM, Rodríguez‐Cabello JC, Arias FJ. Elastin‐like recombinamers: biosynthetic strategies and biotechnological applications. Biotechnol J. 2011;6:1174–86.
Chilkoti A, Christensen T, MacKay JA. Stimulus responsive elastin biopolymers: applications in medicine and biotechnology. Curr Opin Chem Biol. 2006;10:652–7.
Urry DW, Parker TM, Reid MC, Gowda DC. Biocompatibility of the bioelastic materials, poly (GVGVP) and its γ-irradiation cross-linked matrix: summary of generic biological test results. J Bioact Compat Polym. 1991;6:263–82.
Ibáñez‐Fonseca A, Ramos TL, González de Torre I, Sánchez‐Abarca LI, Muntión S, Arias FJ, et al. Biocompatibility of two model elastin‐like recombinamer‐based hydrogels formed through physical or chemical cross‐linking for various applications in tissue engineering and regenerative medicine. J Tissue Eng Regen Med. 2018;12:e1450–e60.
Urry DW. Entropic elastic processes in protein mechanisms. I. Elastic structure due to an inverse temperature transition and elasticity due to internal chain dynamics. J Protein Chem. 1988;7:1–34.
Urry DW, Luan CH, Parker TM, Gowda DC, Prasad KU, Reid MC, et al. Temperature of polypeptide inverse temperature transition depends on mean residue hydrophobicity. J Am Chem Soc. 1991;113:4346–8.
Urry DW, Gowda DC, Parker TM, Luan CH, Reid MC, Harris CM, et al. Hydrophobicity scale for proteins based on inverse temperature transitions. Biopolymers. 1992;32:1243–50.
Girotti A, Reguera J, Rodríguez-Cabello JC, Arias FJ, Alonso M, Testera AM. Design and bioproduction of a recombinant multi (bio) functional elastin-like protein polymer containing cell adhesion sequences for tissue engineering purposes. J Mater Sci Mater Med. 2004;15:479–84.
Wang F, Li Y, Shen Y, Wang A, Wang S, Xie T. The functions and applications of RGD in tumor therapy and tissue engineering. Int J Mol Sci. 2013;14:13447–62.
Ruoslahti E. RGD and other recognition sequences for integrins. Annu Rev Cell Dev Biol. 1996;12:697–715.
D’Souza SE, Ginsberg MH, Plow EF. Arginyl-glycyl-aspartic acid (RGD): a cell adhesion motif. Trends Biochem Sci. 1991;16:246–50.
Heilshorn SC, DiZio KA, Welsh ER, Tirrell DA. Endothelial cell adhesion to the fibronectin CS5 domain in artificial extracellular matrix proteins. Biomaterials. 2003;24:4245–52.
Massia SP, Hubbell JA. Vascular endothelial cell adhesion and spreading promoted by the peptide REDV of the IIICS region of plasma fibronectin is mediated by integrin alpha 4 beta 1. J Biol Chem. 1992;267:14019–26.
Hubbell JA, Massia SP, Desai NP, Drumheller PD. Endothelial cell-selective materials for tissue engineering in the vascular graft via a new receptor. Nat Biotechnol. 1991;9:568.
Papavasiliou G, Cheng M-H, Brey EM. Strategies for vascularization of polymer scaffolds. J Investig Med. 2010;58:838–44.
Kaully T, Kaufman-Francis K, Lesman A, Levenberg S. Vascularization—the conduit to viable engineered tissues. Tissue Eng Part B Rev. 2009;15:159–69.
El-Sherbiny IM, Yacoub MH. Hydrogel scaffolds for tissue engineering: progress and challenges. Glob Cardiol Sci Pract. 2013;3:316–42.
Testa U, Pannitteri G, Condorelli GL. Vascular endothelial growth factors in cardiovascular medicine. J Cardiovasc Med. 2008;9:1190–221.
Lee K, Silva EA, Mooney DJ. Growth factor delivery-based tissue engineering: general approaches and a review of recent developments. J R Soc Interface. 2011;8:153–70.
D’Andrea LD, Iaccarino G, Fattorusso R, Sorriento D, Carannante C, Capasso D, et al. Targeting angiogenesis: structural characterization and biological properties of a de novo engineered VEGF mimicking peptide. Proc Natl Acad Sci. 2005;102:14215–20.
Diana D, Ziaco B, Colombo G, Scarabelli G, Romanelli A, Pedone C, et al. Structural determinants of the unusual helix stability of a de novo engineered vascular endothelial growth factor (VEGF) mimicking peptide. Chem Eur J. 2008;14:4164–6.
Ziaco B, Diana D, Capasso D, Palumbo R, Celentano V, Di Stasi R, et al. C-terminal truncation of vascular endothelial growth factor mimetic helical peptide preserves structural and receptor binding properties. Biochem Biophys Res Commun. 2012;424:290–4.
Wang L, Zhao M, Li S, Erasquin UJ, Wang H, Ren L, et al. “Click” immobilization of a VEGF-mimetic peptide on decellularized endothelial extracellular matrix to enhance angiogenesis. ACS Appl Mater & Interfaces. 2014;6:8401–6.
Leslie-Barbick JE, Saik JE, Gould DJ, Dickinson ME, West JL. The promotion of microvasculature formation in poly (ethylene glycol) diacrylate hydrogels by an immobilized VEGF-mimetic peptide. Biomaterials. 2011;32:5782–9.
Cai L, Dinh CB, Heilshorn SC. One-pot synthesis of elastin-like polypeptide hydrogels with grafted VEGF-mimetic peptides. Biomater Sci. 2014;2:757–65.
de Torre IG, Santos M, Quintanilla L, Testera A, Alonso M, Cabello JCR. Elastin-like recombinamer catalyst-free click gels: characterization of poroelastic and intrinsic viscoelastic properties. Acta Biomater. 2014;10:2495–505.
Staubli SM, Cerino G, De Torre IG, Alonso M, Oertli D, Eckstein F, et al. Control of angiogenesis and host response by modulating the cell adhesion properties of an Elastin-Like Recombinamer-based hydrogel. Biomaterials. 2017;135:30–41.
Nakatsu MN, Sainson RC, Pérez-del-Pulgar S, Aoto JN, Aitkenhead M, Taylor KL, et al. VEGF 121 and VEGF 165 regulate blood vessel diameter through vascular endothelial growth factor receptor 2 in an in vitro angiogenesis model. Lab Invest. 2003;83:1873.
Fischer AH, Jacobson KA, Rose J, Zeller R. Hematoxylin and eosin staining of tissue and cell sections. Cold Spring Harb Protoc. 2008;2008:prot4986. pdb
Fernández-Colino A, Wolf F, Keijdener H, Rütten S, Schmitz-Rode T, Jockenhoevel S, et al. Macroporous click-elastin-like hydrogels for tissue engineering applications. Mater Sci Eng C. 2018;88:140–7.
Testera AM, Girotti A, de Torre IG, Quintanilla L, Santos M, Alonso M, et al. Biocompatible elastin-like click gels: design, synthesis and characterization. J Mater Sci Mater Med. 2015;26:105.
Bishop ET, Bell GT, Bloor S, Broom I, Hendry NF, Wheatley DN. An in vitro model of angiogenesis: basic features. Angiogenesis. 1999;3:335–44.
Ilan N, Cheung L, Pinter E, Madri JA. Platelet-endothelial cell adhesion molecule-1 (CD31), a scaffolding molecule for selected catenin family members whose binding is mediated by different tyrosine and serine/threonine phosphorylation. J Biol Chem. 2000;275:21435–43.
Abhinand CS, Raju R, Soumya SJ, Arya PS, Sudhakaran PR. VEGF-A/VEGFR2 signaling network in endothelial cells relevant to angiogenesis. J Cell Commun Signal. 2016;10:347–54.
D’Andrea LD, De Rosa L, Vigliotti C, Cataldi M. VEGF mimic peptides: Potential applications in central nervous system therapeutics. New Horiz Transl Med. 2017;3:233–51.
Finetti F, Basile A, Capasso D, Di Gaetano S, Di Stasi R, Pascale M, et al. Functional and pharmacological characterization of a VEGF mimetic peptide on reparative angiogenesis. Biochem Pharmacol. 2012;84:303–11.
Chan TR, Stahl PJ, Yu SM. Matrix‐bound vegf mimetic peptides: design and endothelial‐cell activation in collagen scaffolds. Adv Funct Mater. 2011;21:4252–62.
Rodríguez-Cabello JC, Martín L, Alonso M, Arias FJ, Testera AM. “Recombinamers” as advanced materials for the post-oil age. Polym (Guildf). 2009;50:5159–69.
Richards M, Mellor H. In vitro coculture assays of angiogenesis. Methods Mol Biol. 2016;1430:159–66.
Raza F, Zafar H, Zhu Y, Ren Y, Ullah A, Khan AU, et al. A review on recent advances in stabilizing peptides/proteins upon fabrication in hydrogels from biodegradable polymers. Pharmaceutics. 2018;10:16.
Martino MM, Brkic S, Bovo E, Burger M, Schaefer DJ, Wolff T, et al. Extracellular matrix and growth factor engineering for controlled angiogenesis in regenerative medicine. Front Bioeng Biotechnol. 2015;3:45.
Arroyo AG, Iruela-Arispe ML. Extracellular matrix, inflammation, and the angiogenic response. Cardiovasc Res. 2010;86:226–35.
The authors are grateful for the funding from the European Commission (NMP-2014-646075, PITN-GA-2012-317306), MINECO of the Spanish Government (PCIN-2015-010, MAT2015-68901-R, MAT2016-78903-R), Junta de Castilla y León (VA015U16) and Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León.
Conflict of interest
The authors declare that they have no conflict of interest.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Flora, T., de Torre, I.G., Alonso, M. et al. Tethering QK peptide to enhance angiogenesis in elastin-like recombinamer (ELR) hydrogels. J Mater Sci: Mater Med 30, 30 (2019). https://doi.org/10.1007/s10856-019-6232-z