(567f) Protein and Mineral Composition of An Extracellular Matrix Scaffold for Osteogenic Differentiation of Bone Marrow Stromal Cells

Previous studies have demonstrated that electrospun poly(ε-caprolactone) scaffolds can be coated with an osteogenic extracellular matrix generated by bone marrow stromal cells (PCL/ECM scaffold) under engineered conditions.  The PCL/ECM scaffolds have been shown to retain the osteogenic differentiation of bone marrow stromal cells cultured without an osteogenic differentiation factor.  Although the extracellular matrix is known to contain collagen and a calcium-based mineral, the presence of other proteins found in minor quantities in mature bone has not been elucidated.  This research investigated the composition and temporal deposition of the proteins and minerals of osteogenic extracellular matrix scaffolds.  PCL/ECM scaffolds with different extracellular matrix maturities were generated utilizing a flow perfusion bioreactor and analyzed for their protein and mineral composition by liquid chromatography-tandem mass spectroscopy (LC-MS/MS), energy dispersive x-ray diffraction (EDX), x-ray diffraction (XRD), and calcium and phosphate assays.  The analysis revealed that at short culture durations cellular adhesion proteins were deposited, while at longer durations the scaffolds contained collagen 1, hydroxyapatite, matrix remodeling proteins, and regulatory proteins.