Joshua, H Jung
Xiaobin Huang
Tun-Jan Wang
Minh Nguyen
Chider Chen
| Joshua, H Jung | Xiaobin Huang | Tun-Jan Wang | Minh Nguyen | Chider Chen |
Joshua H Jung1, Xiaobin Huang1, Tun-Jan Wang2, Minh Nguyen1, Chider Chen1
1Oral & Maxillofacial Surgery, University of Pennsylvania, School of Dental Medicine 2Periodontics, University of Pennsylvania, School of Dental Medicine
Introduction
Leukocyte-Plate Rich Fibrin (L-PRF) is widely used in clinical dental setting as a biofilm graft material. In such cases, L-PRF usage is associated with shortening healing time and accelerating tissue regeneration in many facets of the oral cavity. However, mechanistic effects of L-PRF are poorly studied and need to be explored. As stem cells from the apical papilla (SCAP) have been shown with superior abilities to survive and differentiate into mineralized tissues even under inflammatory conditions, in this study, we used SCAP as a stem cell model to explore the regulatory mechanisms of L-PRF in bone tissue regeneration.
Methods
SCAP were harvested from healthy patients following institutional review board guidelines. L-PRF prepared using Choukroun’s protocol was used to treat SCAP for 48 hours. RNA-sequencing (RNA-seq) analysis was performed to profile differential gene expression after L-PRF treatment. Stem cell self-renewal assays were performed by Ki67 immunostaining and quantitative PCR (qPCR) analysis. In addition, in vitro osteogenesis and chondrogenesis were accessed by Alizarin Red and immunostaining. Activation of differentiation markers were evaluated by qPCR analysis. To reveal the underlying mechanisms of L-PRF in governing SCAP stemness, pharmaceutical approaches were used to manipulate stem cell function.
Results
RNA-seq analysis showed L-PRF treatment greatly promotes stemness of SCAP via activation of stem cell-related markers and epigenetic pathways. In vitro stem cell characterization further confirms that self-renewal and multipotent differentiation capabilities are largely elevated in L-PRF treated SCAP. Mechanistically, L-PRF treatment highly methylates H3K9 histone residues and activates NOTCH signaling-mediated cilium assembly for SCAP osteogenesis. In summary, this study explores a novel histone modification marker as a downstream target of L-PRF to control NOTCH signaling-mediated cilium assembly in SCAP osteogenesis.
Conclusion
L-PRF treatment increases cell-proliferation and multipotent differentiation through histone H3K9 methylation in SCAP.