title: Combined effect of osteopontin and BMP-2 derived peptides grafted to an adhesive hydrogel on osteogenic and vasculogenic differentiation of marrow stromal cells.
abstract: The objective of this work was to investigate the combined effect of grafting the peptide corresponding to amino acid residues 162-168 of osteopontin (OPD peptide) and the peptide corresponding to amino acid residues 73-92 of bone morphogenetic protein-2 (BMP peptide) to an RGD-conjugated inert hydrogel on osteogenic and vasculogenic differentiation of bone marrow stromal (BMS) cells. RGD-conjugated three-dimensional (3D) porous hydrogel scaffolds with well-defined cylindrical pore geometry were produced from sacrificial wax molds fabricated by fused deposition modeling rapid prototyping system. Propargyl acrylate and 4-pentenal were conjugated to the hydrogel for orthogonal grafting of BMP and OPD peptides by click reaction and oxime ligation, respectively. The OPD peptide was grafted by the reaction between aminooxy moiety of aminooxy-mPEG-OPD (mPEG = mini-poly(ethylene glycol)) and the aldehyde moiety in the hydrogel. The BMP peptide was grafted by the reaction between the azide moiety of Az-mPEG-BMP and the propargyl moiety in the hydrogel. The hydrogels seeded with BMS cells were characterized by biochemical, immunocytochemical, and mRNA analyses. Groups included RGD control hydrogel (RGD), RGD and BMP peptides without OPD (RGD+BMP), RGD and BMP peptides with mutant OPD (RGD+BMP+mOPD), and RGD and BMP peptides with OPD (RGD+BMP+OPD) grafted hydrogels. The extent of mineralization of RGD, RGD+BMP, RGD+BMP+mOPD, and RGD+BMP+OPD groups after 28 days was 650 ± 70, 990 ± 30, 850 ± 30, and 1150 ± 40 mg/(mg of DNA), respectively, indicating that the BMP and OPD peptides enhanced osteogenic differentiation of the BMS cells. The BMS cells seeded on RGD+BMP+OPD grafted hydrogels stained positive for vasculogenic markers α-SMA, PECAM-1, and VE-cadherin while the groups without OPD peptide (RGD+BMP and RGD+BMP+mOPD) stained only for α-SMA but not PECAM-1 or VE-cadherin. These results were consistent with the significantly higher PECAM-1 mRNA expression for RGD+BMP+OPD group after 21 and 28 days, compared to the groups without OPD. These findings suggest that the RGD+BMP+OPD peptides provide a favorable microenvironment for concurrent osteogenic and vasculogenic differentiation of progenitor marrow-derived cells.
authors.affiliation: Biomimetic Materials and Tissue Engineering Laboratories, Department of Chemical Engineering, University of South Carolina, Columbia, South Carolina 29208, USA.
authors.forename: Xuezhong
authors.initials: X
authors.lastname: He
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Yang
authors.forename: Esmaiel
authors.initials: E
authors.lastname: Jabbari
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001672:Biocompatible Materials; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000008:administration & dosage*; D015820:Cadherins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D020100:Hydrogels; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000235:genetics; D009842:Oligopeptides / Q000008:administration & dosage; D015675:Osteocalcin / Q000235:genetics; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D015676:Osteonectin / Q000235:genetics; D053495:Osteopontin / Q000008:administration & dosage* / Q000235:genetics; D010446:Peptide Fragments / Q000008:administration & dosage; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D011091:Polyesters; D011092:Polyethylene Glycols; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017154:Stromal Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D054457:Tissue Scaffolds; D000099152:Cadherin 5
chemical_list: D000199:Actins; D015703:Antigens, CD; D001672:Biocompatible Materials; D055396:Bone Morphogenetic Protein 2; D015820:Cadherins; D020100:Hydrogels; D009842:Oligopeptides; D015675:Osteocalcin; D015676:Osteonectin; D053495:Osteopontin; D010446:Peptide Fragments; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011091:Polyesters; D011092:Polyethylene Glycols; D012333:RNA, Messenger; D000099152:Cadherin 5; C525720:Bmp2 protein, rat; C506897:Spp1 protein, rat; C512585:poly(L-lactide-co-ethylene oxide-co-fumarate); C498862:Acta2 protein, rat; C047981:arginyl-glycyl-aspartic acid