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DAR 16-II Primes Endothelial Cells for Angiogenesis Improving Bone Ingrowth in 3D-Printed BCP Scaffolds and Regeneration of Critically Sized Bone Defects

Affiliation
Faculty of Dentistry, Oral Biology Department, King Abdulaziz University, Jeddah 21589, Saudi Arabia
Alfayez, Eman;
Affiliation
Faculty of Dentistry, Oral & Craniofacial Sciences King’s College London, London SE1 9RT, UK
Veschini, Lorenzo;
ORCID
0000-0002-6823-3375
Affiliation
Department of Industrial Engineering, University of Padua, 35131 Padua, Italy
Dettin, Monica;
ORCID
0000-0002-8113-0346
Affiliation
Department of Civil, Environmental, and Architectural Engineering, University of Padua, 35131 Padua, Italy
Zamuner, Annj;
ORCID
0000-0001-5610-0797
Affiliation
Fondazione Ricerca nel Mediterraneo (Ri.MED) and Department of Laboratory Medicine and Advanced Biotechnologies, Istituto di Ricovero e Cura a Carattere Scientifico-Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, 90100 Palermo, Italy
Gaetani, Massimiliano;
ORCID
0000-0002-7389-7555
Affiliation
Fondazione Ricerca nel Mediterraneo (Ri.MED) and Department of Laboratory Medicine and Advanced Biotechnologies, Istituto di Ricovero e Cura a Carattere Scientifico-Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, 90100 Palermo, Italy
Carreca, Anna P.;
ORCID
0000-0002-2411-9802
Affiliation
Faculty of Medicine, University of Niš, 18000 Niš, Serbia
Najman, Stevo;
ORCID
0000-0002-7897-0854
Affiliation
Department for Oral, Cranio-Maxillofacial and Facial Plastic Surgery, Medical Center of the Goethe University, 60323 Frankfurt, Germany
Ghanaati, Shahram;
ORCID
0000-0001-7122-7980
Affiliation
Faculty of Dentistry, Oral & Craniofacial Sciences King’s College London, London SE1 9RT, UK
Coward, Trevor;
Affiliation
Faculty of Dentistry, Oral & Craniofacial Sciences King’s College London, London SE1 9RT, UK
Di Silvio, Lucy

Bone is a highly vascularized tissue and relies on the angiogenesis and response of cells in the immediate environmental niche at the defect site for regeneration. Hence, the ability to control angiogenesis and cellular responses during osteogenesis has important implications in tissue-engineered strategies. Self-assembling ionic-complementary peptides have received much interest as they mimic the natural extracellular matrix. Three-dimensional (3D)-printed biphasic calcium phosphate (BCP) scaffolds coated with self-assembling DAR 16-II peptide provide a support template with the ability to recruit and enhance the adhesion of cells. In vitro studies demonstrated prompt the adhesion of both human umbilical vein endothelial cells (HUVEC) and human mesenchymal stem cells (hMSC), favoring endothelial cell activation toward an angiogenic phenotype. The SEM-EDS and protein micro bicinchoninic acid (BCA) assays demonstrated the efficacy of the coating. Whole proteomic analysis of DAR 16-II-treated HUVECs demonstrated the upregulation of proteins involved in cell adhesion (HABP2), migration (AMOTL1), cytoskeletal re-arrangement (SHC1, TMOD2), immuno-modulation (AMBP, MIF), and morphogenesis (COL4A1). In vivo studies using DAR-16-II-coated scaffolds provided an architectural template, promoting cell colonization, osteogenesis, and angiogenesis. In conclusion, DAR 16-II acts as a proactive angiogenic factor when adsorbed onto BCP scaffolds and provides a simple and effective functionalization step to facilitate the translation of tailored 3D-printed BCP scaffolds for clinical applications.

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