title: Creation of bony microenvironment with CaP and cell-derived ECM to enhance human bone-marrow MSC behavior and delivery of BMP-2.
abstract: Extracellular matrix (ECM) comprises a rich meshwork of proteins and proteoglycans, which not only contains biological cues for cell behavior, but is also a reservoir for binding growth factors and controlling their release. Here we aimed to create a suitable bony microenvironment with cell-derived ECM and biodegradable β-tricalcium phosphate (β-TCP). More specifically, we investigated whether the ECM produced by bone marrow-derived mesenchymal stem cells (hBMSC) on a β-TCP scaffold can bind bone morphogenetic protein-2 (BMP-2) and control its release in a sustained manner, and further examined the effect of ECM and the BMP-2 released from ECM on cell behaviors. The ECM was obtained through culturing the hBMSC on a β-TCP porous scaffold and performing decellularization and sterilization. SEM, XPS, FTIR, and immunofluorescent staining results indicated the presence of ECM on the β-TCP and the amount of ECM increased with the incubation time. BMP-2 was loaded onto the β-TCP with and without ECM by immersing the scaffolds in the BMP-2 solution. The loading and release kinetics of the BMP-2 on the β-TCP/ECM were significantly slower than those on the β-TCP. The β-TCP/ECM exhibited a sustained release profile of the BMP-2, which was also affected by the amount of ECM. This is probably because the β-TCP/ECM has different binding mechanisms with BMP-2. The β-TCP/ECM promoted cell proliferation. Furthermore, the BMP-2-loaded β-TCP/ECM stimulated reorganization of the actin cytoskeleton, increased expression of alkaline phosphatase and calcium deposition by the cells compared to those without BMP-2 loading and the β-TCP with BMP-2 loading.
authors.affiliation: Department of Restorative Dentistry and Biomaterials, The University of Texas Health Science Center at Houston, 6516 M.D. Anderson Blvd., Houston, TX 77030, USA.
authors.forename: Yunqing
authors.initials: Y
authors.lastname: Kang
authors.forename: Sungwoo
authors.initials: S
authors.lastname: Kim
authors.forename: Ali
authors.initials: A
authors.lastname: Khademhosseini
authors.forename: Yunzhi
authors.initials: Y
authors.lastname: Yang
mesh_terms: D055396:Bone Morphogenetic Protein 2 / Q000008:administration & dosage*; D002130:Calcium Phosphates / Q000737:chemistry*; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000737:chemistry*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D056951:Photoelectron Spectroscopy; D017550:Spectroscopy, Fourier Transform Infrared; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D055396:Bone Morphogenetic Protein 2; D002130:Calcium Phosphates; C485817:beta-tricalcium phosphate