Marc Martínez-Miguel, Miquel Castellote-Borrell, Mariana Köber, Adriana R. Kyvik, Judit Tomsen-Melero, Guillem Vargas-Nadal, Jose Muñoz, Daniel Pulido, Edgar Cristóbal-Lecina, Solène Passemard, Miriam Royo, Marta Mas-Torrent, Jaume Veciana, Marina I. Giannotti, Judith Guasch, Nora Ventosa, Imma Ratera, Ministerio de Ciencia, Innovación y Universidades (España), Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red Bioingeniería, Biomateriales y Nanomedicina (España), Generalitat de Catalunya, Fundació La Marató de TV3, European Cooperation in Science and Technology, Max Planck Society, European Commission, Köber, Mariana, Kyvik Ruiz, Adriana, Tomsen Melero, Judit, Pulido, Daniel, Royo, Miriam, Mas Torrent, Marta, Veciana, Jaume, Giannotti, Marina I., Guasch, Judith, Ventosa, Nora, Ratera, Imma, Köber, Mariana [0000-0001-9962-7900], Kyvik Ruiz, Adriana [0000-0002-6385-7162], Tomsen Melero, Judit [0000-0002-2837-8107], Pulido, Daniel [0000-0002-2841-194X], Royo, Miriam [0000-0001-5292-0819], Mas Torrent, Marta [0000-0002-1586-005X], Veciana, Jaume [0000-0003-1023-9923], Giannotti, Marina I. [0000-0002-0815-742X], Guasch, Judith [0000-0002-3571-4711], Ventosa, Nora [0000-0002-8008-4974], and Ratera, Imma [0000-0002-1464-9789]
The synthesis and study of the tripeptide Arg-Gly-Asp (RGD), the binding site of different extracellular matrix proteins, e.g., fibronectin and vitronectin, has allowed the production of a wide range of cell adhesive surfaces. Although the surface density and spacing of the RGD peptide at the nanoscale have already shown a significant influence on cell adhesion, the impact of its hierarchical nanostructure is still rather unexplored. Accordingly, a versatile colloidal system named quatsomes, based on fluid nanovesicles formed by the self-assembling of cholesterol and surfactant molecules, has been devised as a novel template to achieve hierarchical nanostructures of the RGD peptide. To this end, RGD was anchored on the vesicle's fluid membrane of quatsomes, and the RGD-functionalized nanovesicles were covalently anchored to planar gold surfaces, forming a state of quasi-suspension, through a long poly(ethylene glycol) (PEG) chain with a thiol termination. An underlying self-assembled monolayer (SAM) of a shorter PEG was introduced for vesicle stabilization and to avoid unspecific cell adhesion. In comparison with substrates featuring a homogeneous distribution of RGD peptides, the resulting hierarchical nanoarchitectonic dramatically enhanced cell adhesion, despite lower overall RGD molecules on the surface. The new versatile platform was thoroughly characterized using a multitechnique approach, proving its enhanced performance. These findings open new methods for the hierarchical immobilization of biomolecules on surfaces using quatsomes as a robust and novel tissue engineering strategy., This work was supported by MICINN (PID2019-105622RBI00, MAT2016-80826-R, PID2019-111682RB-I00, PID2020-115296RA-I00, CTQ2015-66194-R; SAF2014-60138-R, RTI2018-093831-B-I00, and PDC2021-121481-I00); Instituto de Salud Carlos III (ISCIII) through the Networking Research Center on Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN (FlexQS-skin, FlexCAB, BBN18PI01, BBN20PIV02, and CB/06/0074); Generalitat de Catalunya (grants 2017-SGR-918, 2017-SGR-229, 2017-SGR-1442, 2017-SGR-1439); the Fundació Marató de TV3 (Nr. 201812); the COST Action CA15126 Between Atom and Cell, and “ERDF A way of making Europe”. J.G. acknowledges financial support from the Ramón y Cajal Program (RYC-2017-22614) from MICINN and the Max Planck Society through the Max Planck Partner Group “Dynamic Biomimetics for Cancer Immunotherapy” in collaboration with the Max Planck Institute for Medical Research (Heidelberg, Germany). This work has received funding from the European Union’s Horizon 2020 research and innovation program through grant agreements 953110 (PHOENIX), 720942 (Smart4Fabry), 101007804 (MICRO4NANO), and 801342 (granted to the Agency for Business Competitiveness ACCIÓ through a Tecniospring Industry fellowship (TECSPR19-1-0065)). ICMAB acknowledges support from MICINN through the “‘Severo Ochoa”’ Programme for Centres of Excellence in R&D (CEX2019-000917-S). J.M. acknowledges a “Juan de la Cierva” fellowship from MICINN. J.T-M. acknowledges an FI-AGAUR grant (2020FI_B2 00137) from Generalitat de Catalunya and the European Social Fund. We also acknowledge the ICTS “NANBIOSIS for the support of the Synthesis of Peptides Unit (U3) at IQAC–CSIC (https://www.nanbiosis.es/portfolio/u3-synthesis-of-peptides-unit/) and the Biomaterial Processing and Nanostructuring Unit (U6) at ICMAB-CSIC (https://www.nanbiosis.es/portfolio/u6-biomaterial-processing-and-nanostructuring-unit/). We are grateful to the SMP unit of the Scientific and Technological Centers of University of Barcelona (CCiTUB). This work has been developed under the “Biochemistry, Molecular Biology and Biomedicine” and “Materials Science” Ph.D. programs of Universitat Autònoma de Barcelona (UAB)., With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).