In situ delivery of BMP-2 and VEGF for osteogenesis of mesenchymal stem cells and bone regeneration in rat calvarial defect model
- In situ delivery of BMP-2 and VEGF for osteogenesis of mesenchymal stem cells and bone regeneration in rat calvarial defect model
- 라메쉬; 반세영; 두 핑; 노용권; 김인걸; 황민태; 박귀덕
- Core-shell microcapsules; BMP-2; VEGF; umbilical cord-derived mesenchymal stem cells; Osteogenesis; Rat calvarial model
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- In this study, we investigate the effect of in situ dual growth factors (GFs) delivery for osteogenic differentiation of umbilical cord derived mesenchymal stem cells (UC-MSCs) and for bone regeneration in rat calvarial defect model. Ultrasonication-assisted emulsification produces bone morphogenic protein (BMP)-2 loaded poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs; ~600 nm size), whereas electrodropping technique based on custom-made coaxial needles yields vascular endothelial growth factor (VEGF) loaded alginate beads (~100 µm). For dual GFs delivery, PLGA NPs are homogeneously encapsulated within alginate beads that results in core-shell MCs (C-S MCs) formation. In vitro release test proves the sequential delivery of dual GFs from C-S MCs over a period of 28 days. Mixture of UC-MSCs and collagen is used to evaluated MSCs osteogenesis by comparing positive control (G1: BMP-2 in medium) with single GF (G2: BMP-2) or dual GF MCs (G3: BMP-2/VEGF) encapsulation. The PKH-26 labeled MSCs reveal that MCs and UC-MSCs are uniformly distributed within collagen scaffold. The in vitro osteogenesis of UC-MSCs (28 days) is confirmed by von Kossa staining and immunofluorescence (osteocalcin). In addition, quantitative analysis of calcium content, alkaline phosphatase (ALP) activity, and osteogenic gene expression further demonstrate the positive effect of dual GF carrying MCs in the induction of MSCs osteogenesis. Meanwhile further study shows a significant improvement of bone regeneration in rat calvarial model (8 mm diameter) using dual GF delivery MCs. In summary, C-S MCs-mediated dual GF delivery proves feasible and suggests its osteogenic potential with UC-MSCs as well as with bone defect model.
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