Abstract
Platelet-rich plasma (PRP) has a pool of multiple growth factors efficient at inducing the proliferation and osteogenic differentiation of human adipose-derived stem cells (hADSCs). Bone morphogenetic protein (BMP)-2 is a strong stimulator for the osteogenic differentiation of hADSCs. The purpose of this study was to verify the effect of PRP-released growth factors and microsphere-encapsulated BMP-2 on the proliferation and osteoblastic differentiation of hADSCs and to construct a novel tissue-engineered bone. The BMP-2-loaded microspheres and hADSCs were embedded in activated PRP gel. Another 5 composites (hADSCs/platelet-poor plasma [PPP]; hADSCs/PRP; hADSCs/BMP-2/PPP; hADSCs/BMP-2/PRP; and hADSCs/BMP-2+microspheres/PPP) were also constructed. The DNA content, alkaline phosphatase activity, mRNA expression of alkaline phosphatase, osteopontin, osteocalcin, and mineralization of hADSCs in each composite were compared. The DNA content was higher in all PRP-containing composites, meaning that PRP-released growth factors stimulated proliferation of hADSCs. Alkaline phosphatase increased in BMP-2/PRP and BMP-2+microspheres/PRP composites in the first 7 days, meaning that BMP-2 had a synergistic effect with PRP in the early differentiation of hADSCs. Osteopontin, osteocalcin, and mineralization assays were higher in BMP-2+microspheres/PRP composite than in the BMP-2/PRP composite up to 21 days, meaning that a continuous delivery of BMP-2 stimulates osteoblastic differentiation of hADSCs at the early stage and the final maturation stage. These results suggest that sustained delivery of BMP-2 in combination with PRP is better than a single administration of PRP or BMP-2 in the osteogenic differentiation of hADSCs.
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