The Role of Platelet-Rich Plasma on the Chondrogenic and Osteogenic Differentiation of Human Amniotic-Fluid-Derived Stem Cells

Abstract

Amniotic fluid represents a new and promising source of engraftable stem cells. The purpose of this study was to investigate the in vitro effects of platelet-rich plasma (PRP) on amniotic-fluid-derived stem cells (AFSCs) on chondrogenic or osteogenic differentiation potential. Amniotic fluid samples were obtained from women undergoing amniocentesis for prenatal diagnosis at 16-18 weeks of pregnancy. Undifferentiated human AFSCs were cocultured with PRP for 14 days. The study includes two protocols investigating the effects of activated PRP using two different methods: via freeze-thaw cycles and via the addition of calcium gluconate. On the 14th day of culturing, the differentiation potential of the cocultured AFSCs was then compared with undifferentiated AFSCs. Staining with alcian blue solution (ABS) and alizarine red solution (ARS) was performed, and chondrogenic- and osteogenic-associated genes markers were investigated. ABS demonstrated enhanced glycosaminoglycan expression. Cocultured cells expressed chondrocyte-associated genes, determined by real-time polymerase chain reaction (RT-PCR), including type I collagen, type II collagen, COMP, and aggrecan. In regard to the osteogenic markers, osteopontin and bone sialoprotein, there were no changes. In particular, the activation of PRP using the freeze-thaw cycle protocol showed a higher expression of the chondrogenic markers. Our preliminary in vitro results showed that PRP has good potential in the chondrogenic differentiation of AFSCs.