Osteogenesis of human placenta-derived mesenchymal stem cell using biomimetic polymer mesh scaffold coated with BMP-2 tethered fibroblast derived matrix
- Osteogenesis of human placenta-derived mesenchymal stem cell using biomimetic polymer mesh scaffold coated with BMP-2 tethered fibroblast derived matrix
- 김인걸; 두 핑; 고재훈; 하철원; 박귀덕
- Extracellular matrix; Microenvironment; Placenta-derived mesenchymal stem cell; Mesh scaffold; Osteogenesis
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- The objective of this study is to construct a biomimetic composite by combining PLGA/PLA blended mesh scaffold with bone morphogenic protein (BMP-2) loaded cell-derived extracellular matrix (CDM) to promote bone formation. Decellularized ECM from in vitro cultured human lung fibroblast (hFDM) was coated onto the surface of mesh scaffolds. Heparin was then conjugated on the surface of ECM-coated mesh scaffold using EDC chemistry and BMP-2 was subsequently loaded onto it. The sustained release of BMP-2 was observed for 28 days. Human placenta-derived mesenchymal stem cells (hPMSCs) were seeded into hFDM-coated mesh scaffolds and cultured for 7, 14, and 28 days in either growth or osteogenic medium. Cells were growing attached to the microfiber structure and proliferated better in hFDM-coated mesh as determined by environmental SEM and CCK-8 assay, respectively. When osteogenic differentiation of the hPMSCs was determined by using alkaline phosphatase (ALP) activity and osteogenic marker expression, both parameters were more upregulated with hFDM-coated mesh group than those of control and fbronectin (FN)-coated group. Meanwhile, hPMSCs-seeded mesh scaffolds were also transplanted in the subcutaneous region of nude mice. Histological findings demonstrated that BMP-2 tethered hFDM-coated mesh scaffold significantly increased microvessel density and promoted the formation of a mineralized matrix, as compared to FN or hFDM group. In conclusion, BMP-2 tethered, bioactive ECM-coated mesh scaffold is a very effective microenvironment for osteogenesis and therefore can be an excellent candidate for bone tissue engineering.
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