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  • Material-driven fibronectin and vitronectin assembly enhances BMP-2 presentation and osteogenesis.

Material-driven fibronectin and vitronectin assembly enhances BMP-2 presentation and osteogenesis.

Materials today. Bio (2022-08-09)
Yinbo Xiao, Hannah Donnelly, Mark Sprott, Jiajun Luo, Vineetha Jayawarna, Leandro Lemgruber, P Monica Tsimbouri, R M Dominic Meek, Manuel Salmeron-Sanchez, Matthew J Dalby
ABSTRACT

Mesenchymal stem cell (MSC)-based tissue engineering strategies are of interest in the field of bone tissue regenerative medicine. MSCs are commonly investigated in combination with growth factors (GFs) and biomaterials to provide a regenerative environment for the cells. However, optimizing how biomaterials interact with MSCs and efficiently deliver GFs, remains a challenge. Here, via plasma polymerization, tissue culture plates are coated with a layer of poly (ethyl acrylate) (PEA), which is able to spontaneously permit fibronectin (FN) to form fibrillar nanonetworks. However, vitronectin (VN), another important extracellular matrix (ECM) protein forms multimeric globules on the polymer, thus not displaying functional groups to cells. Interestingly, when FN and VN are co-absorbed onto PEA surfaces, VN can be entrapped within the FN fibrillar nanonetwork in the monomeric form providing a heterogeneous, open ECM network. The combination of FN and VN promote MSC adhesion and leads to enhanced GF binding; here we demonstrate this with bone morphogenetic protein-2 (BMP2). Moreover, MSC differentiation into osteoblasts is enhanced, with elevated expression of osteopontin (OPN) and osteocalcin (OCN) quantified by immunostaining, and increased mineralization observed by von Kossa staining. Osteogenic intracellular signalling is also induced, with increased activity in the SMAD pathway. The study emphasizes the need of recapitulating the complexity of native ECM to achieve optimal cell-material interactions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bone Morphogenetic Protein 2 human, BMP-2, recombinant, expressed in HEK 293 cells, HumanKine, suitable for cell culture
Sigma-Aldrich
Fibronectin human plasma, lyophilized powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Anti-Vinculin antibody, Mouse monoclonal, clone hVIN-1, purified from hybridoma cell culture