Allogeneic Fibrin Clot for Odontogenic/Cementogenic Differentiation of Human Dental Mesenchymal Stem Cells

ABSTRACT

BACKGROUND@#Fetal bovine serum is widely used as a growth supplement for cell culture medium; however, animalbornepathogens increase the risk of transmitting infectious agents. Platelet-rich fibrin is recently considered as a successfulalternative but leukocytes present limits to its allogeneic feasibility. The aim of this study was to explore the effects ofallogeneic fibrin clot (AFC) without leukocytes on inducing odontogenic/cementogenic differentiation of human dentalpulp stem cells (hDPSCs) and human periodontal ligament stem cells (hPDLSCs) in vitro and in vivo. @*METHODS@#AFC was prepared by high-speed centrifugation and leukocytes were almost removed, and AFC serum wasobtained through three freeze–thaw cycles. hDPSCs and hPDLSCs were treated with AFC serum to investigate theodontogenic or cementogenic associated markers by real-time polymerase chain reaction. hDPSCs were treated with AFCserum and placed inside of dentin canal, hPDLSCs were treated with AFC serum to wrap outside of dentin, the mixture wasthen transplanted into the subcutaneous of nude mice for 12 weeks. @*RESULTS@#AFC serum exhibited enough growth factors and cytokines to induce odontogenic/cementogenic differentiationof hDPSCs and hPDLSCs in vitro. Furthermore, AFC seurum could induce hDPSCs to differentiate into odontoblastslikecells and pulp-like tissues, and hPDLSCs to regenerate cementum-like tissues. @*CONCLUSION@#AFC could be an alternative safe source with growth factors for the expansion of human dental mesenchymalstem cells (hDMSCs).