Tissue engineering provides an important approach for bone regeneration. Calcium phosphate cement (CPC) can be injected to fill complex-shaped bone defects with excellent osteoconductivity. Induced pluripotent stem cells (iPSCs) are exciting for regenerative medicine due to their potential to proliferate and differentiate into cells of all three germ layers. To date, there has been no report on iPSC seeding with CPC scaffolds. The objectives of this study were to (1) obtain iPSC-derived mesenchymal stem cells (iPSC-MSCs); (2) seed iPSC-MSCs on CPC scaffold for the first time to investigate cell attachment and proliferation; and (3) investigate osteogenic differentiation of iPSC-MSCs on CPC and mineral synthesis by the cells. iPSCs were derived from adult marrow CD34+ cells that were reprogrammed by a single episomal vector pEB-C5. iPSCs were cultured to form embryoid bodies (EBs), and MSCs were migrated out of EBs. Flow cytometry indicated that iPSC-MSCs expressed typical surface antigen profile of MSCs. Mesenchymal differentiation of iPSC-MSCs demonstrated that the iPSC-MSCs had the potential to differentiate into adipocytes, chondrocytes, and osteoblasts. iPSC-MSCs had good viability when attached on CPC scaffold. iPSC-MSCs differentiated into the osteogenic lineage and synthesized bone minerals. iPSC-MSCs on CPC in osteogenic medium yielded higher gene expressions of osteogenic markers including alkaline phosphatase (ALP), osteocalcin, collagen type I, and Runt-related transcription factor 2 than those in control medium (p<0.05). iPSC-MSCs on CPC in osteogenic medium had 10-fold increase in ALP protein than that in control medium (p<0.05). Bone mineral synthesis by iPSC-MSCs adherent to CPC scaffold was increased with time, and mineralization in osteogenic medium was three to four fold that in control medium. In conclusion, iPSCs were derived from adult marrow CD34+ cells that were reprogrammed by a single episomal vector pEB-C5, and MSCs were generated from the EBs. iPSC-MSCs showed good viability and osteogenic differentiation on CPC scaffold for the first time; hence, the novel iPSC-MSC-CPC construct is promising to promote bone regeneration in dental, craniofacial, and orthopedic repairs.
ASJC Scopus subject areas
- Biomedical Engineering