Regenerative Potential of Dental Pulp Cells Cultured Under Inflammatory Conditions Using Platelet-Rich Plasma,
Diduo Tian1, Yunjie Shuai1, Yan Wei1
1State Key Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Introduction:
The development of various autologous platelet concentrates (APCs) has garnered recent attention for various applications in tissue regeneration. The aim of this study was to investigate 3 protocols to produce APCs on the biological capacity of human dental pulp cells (hDPCs) cultured under both normal and inflammatory conditions (induced by LPS from E. coli) in vitro.
Methods:
HDPCs were cultured in culture media from either (1) platelet-rich plasma (first centrifugation at 900 RCF for 5 minutes and second centrifugation at 2000 RCF for 15 minutes), (2) injectable-platelet-rich fibrin (700 RPM for 3 minutes), or (3) concentrated platelet-rich fibrin (C-PRF) (2000 RCF for 8 minutes) when cultured under normal and inflammatory conditions. Cell migration was assessed using a scratch and transwell assay. Cell proliferation was tested using EdU assay and Ki67 immunofluorescence. HDPCs differentiation was assessed via Alizarin Red Staining, DSPP staining and genes encoding dentin matrix protein 1, dentin sialophosphoprotein, and collagen type I alpha 1. Additionally, hDPCs conditioned under an inflammatory condition were further monitored for genes encoding P65 and interleukin 1 beta.
Results:
All APC groups demonstrated the ability to promote migration and proliferation with C-PRF demonstrating significantly highest values. Platelet-rich plasma and injectable-platelet-rich fibrin moderately increased mineralized nodule formation and odontogenic marker expression, whereas C-PRF showed the strongest enhancement of Alizarin Red staining and the expression of dentin matrix protein 1, dentin sialophosphoprotein, and collagen type I alpha 1. While culture conditions under inflammatory conditions induced by LPS induced higher interleukin 1 beta, P65, and lower hDPCs differentiation, the use of APCs improved all outcomes and the C-PRF group showed the greatest reduction in negative effects caused by LPS/inflammation.
Conclusions:
Based on these findings, the use of APCs was able to improve hDPCs activity in vitro with the C-PRF group showing the best results. Noteworthy, while LPS conditioned media typically led to higher hDPCs inflammation, the use of each APC group, especially C-PRF, was able to attenuate some of this impact. Future large animal and clinical studies are needed to further validate these findings.
