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The need for bone tissue engineering has increased as the world population ages. The objectives of this study were to (1) develop a novel human umbilical cord mesenchymal stem cell (hUCMSC)-encapsulating, fiber-reinforced injectable calcium phosphate cement (CPCF) scaffold, and (2) investigate the effects of osteogenic media delivery, preosteodifferentiation, and bone morphogenetic protein-2 (BMP-2) delivery on hUCMSC osteodifferentiation inside CPCF for the first time. CPCF was developed using calcium phosphate powders, chitosan, and absorbable fibers. Four types of hUCMSC-encapsulating constructs were fabricated: control media in alginate hydrogel microbeads in CPCF; osteogenic media in microbeads; preosteodifferentiation; and recombinant human BMP-2 (rhBMP-2) in microbeads. The hUCMSCs inside CPCF maintained good viability, successfully differentiated into the osteogenic lineage, and synthesized bone minerals. The preosteodifferentiation method yielded high gene expressions of alkaline phosphatase, osteocalcin, collagen, and osterix, as well as alkaline phosphatase protein synthesis. The mineralization for the preosteodifferentiation constructs exceeded those of the rhBMP-2 group at 1-7 days, and was slightly lower than the rhBMP-2 group at 21 days. Mineralization of the rhBMP-2 group was 12-fold that of the control constructs at 21 days. In conclusion, although the BMP-2 delivery promoted osteodifferentiation, the preosteodifferentiation method and the ostegenic media method with hUCMSCs in CPCF were also promising for bone regeneration. hUCMSCs may be an effective alternative to the gold-standard bone marrow MSCs, which require an invasive procedure to harvest. The novel injectable stem cell-CPCF construct may be useful in minimally invasive and other orthopedic surgeries.

tissue engineering. part a

Osteogenic Media and rhBMP-2-Induced Differentiation of Umbilical Cord Mesenchymal Stem Cells Encapsulated in Alginate Microbeads and Integrated in an Injectable Calcium Phosphate-Chitosan Fibrous Scaffold