Background: Fetal bovine serum is widely used as a growth supplement for cell culture medium; however, animal-borne pathogens increase the risk of transmitting infectious agents. Platelet-rich fibrin is recently considered as a successful alternative but leukocytes present limits to its allogeneic feasibility. The aim of this study was to explore the effects of allogeneic fibrin clot (AFC) without leukocytes on inducing odontogenic/cementogenic differentiation of human dental pulp 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 was obtained through three freeze-thaw cycles. hDPSCs and hPDLSCs were treated with AFC serum to investigate the odontogenic or cementogenic associated markers by real-time polymerase chain reaction. hDPSCs were treated with AFC serum and placed inside of dentin canal, hPDLSCs were treated with AFC serum to wrap outside of dentin, the mixture was then transplanted into the subcutaneous of nude mice for 12 weeks.
Results: AFC serum exhibited enough growth factors and cytokines to induce odontogenic/cementogenic differentiation of hDPSCs and hPDLSCs in vitro. Furthermore, AFC seurum could induce hDPSCs to differentiate into odontoblasts-like cells 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 mesenchymal stem cells (hDMSCs).
Keywords: Allogeneic fibrin clot; Human dental mesenchymal stem cells; Leukocyte; Odongtogenic/cementogenic; Platelet-rich fibrin.