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Abstract:
Cartilage engineering is a new therapeutic approach in regenerative medicine. This study explored the chondrogenic potential of human dental pulp stem/stromal cells (hDPSCs) and adipose-derived stem/stromal cells (hAD-MSCs) grown on a hydrolytically modified poly(L-lactide-co-caprolactone) (PLCL) electrospun scaffold in relation to the phenotype of primary chondrocytes on PLCL. The effects of PLCL scaffold on the biological features of hDPSC, hAD-MSC, and their chondrogenic differentiation and chondrocytes biology were evaluated via flow cytometry, immunochemistry, biochemistry, and RT–PCR. The results demonstrated that PLCL supported hDPSC, hAD-MSC, and chondrocyte viability and cellular attachment. The chondrogenic potential of hDPSCs and hAD-MSCs on PLCL scaffold was evidenced by the mRNA expression of the cartilage-specific genes. Collagen type II (Col II) and aggrecan (Acan) gene expression and their proteins significantly increased in chondrogenically differentiated hDPSCs and hAD-MSCs on PLCL compared with undifferentiated stem/stromal cells on PLCL. The phenotype of differentiated hDPSCs and hAD-MSCs was comparable to primary chondrocytes grown on PLCL. The results of this study showed that PLCL scaffold promoted chondrogenic differentiation of hAD-MSCs and hDPSCs toward chondrocytes with phenotypic similarities to native chondrocytes. The PLCL scaffold composition has a positive effect on hDPSC, hAD-MSC, and chondrocyte behavior, chondrogenic gene expression, and matrix protein synthesis.

Keywords:
stem/stromal cells, hDPSCs, hAD-MSCs, chondrocytes, chondrogenesis, poly(L-lactide-co-caprolactone) (PLCL), cartilage engineering

Abstract:
Bone tissue engineering using different scaffolds is a new therapeutic approach in regenerative medicine. This study explored the osteogenic potential of human dental pulp stem cells (hDPSCs) grown on a hydrolytically modified poly(L-lactide-co-caprolactone) (PLCL) electrospun scaffold and a non-woven hyaluronic acid (HYAFF-11™) mesh. The adhesion, immunophenotype, and osteogenic differentiation of hDPSCs seeded on PLCL and HYAFF-11™ scaffolds were analyzed. The results showed that PLCL and HYAFF-11™ scaffolds significantly supported hDPSCs adhesion; however, hDPSCs’ adhesion rate was significantly higher on PLCL than on HYAFF-11™. SEM analysis confirmed good adhesion of hDPSCs on both scaffolds before and after osteogenesis. Alizarin red S staining showed mineral deposits on both scaffolds after hDPSCs osteogenesis. The mRNA levels of runt-related transcription factor 2 (Runx2), collagen type I (Coll-I), osterix (Osx), osteocalcin (Ocn), osteopontin (Opn), bone sialoprotein (Bsp), and dentin sialophosphoprotein (Dspp) gene expression and their proteins were higher in hDPSCs after osteogenic differentiation on both scaffolds compared to undifferentiated hDPSCs on PLCL and HYAFF-11™. These results showed that PLCL scaffolds provide a better environment that supports hDPSCs attachment and osteogenic differentiation than HYAFF-11™. The high mRNA of early osteogenic gene expression and mineral deposits observed after hDPSCs osteogenesis on a PLCL mat indicated its better impact on hDPSCs’ osteogenic potential than that of HYAFF-11™, and hDPSC/PLCL constructs might be considered in the future as an innovative approach to bone defect repair.

Keywords:
dental stem cells, hDPSCs, osteogenesis, PLCL scaffold, HYAFF-11 scaffold