Abstract
Regenerative endodontic procedures using autologous platelet-rich plasma (PRP) can improve the biologic outcome of treatment. However, its mechanism of action on improving pulp regeneration is not fully elucidated. Autophagy was recently shown to be related to tissue repair and osteogenesis. Therefore, the objective of this study was to investigate the effect of PRP in dental pulp regeneration and to elucidate the role of autophagy involved in this process. Human dental pulp cells (hDPCs) were isolated from healthy dental pulp and co-cultured with an increasing concentration of PRP. Cellular migration and proliferation were determined by scratch assay, transwell assay, and cell-counting kit 8 assay. Osteogenic differentiation was clarified by using alkaline phosphatase staining, alizarin red staining, and real-time polymerase chain reaction (RT-PCR) to measure the gene expression levels of alkaline phosphatase, collagen-1, osteocalcin, dentin matrix protein 1, and dentin sialophosphoprotein. Autophagic bodies were observed by transmission electron microscopy and the expression of autophagy marker light chain 3B (LC3B) was determined by immunofluorescence staining. The mRNA and protein expression level of LC3B and Beclin-1 were quantified by qRT-PCR and western blotting. The effect of PRP on cellular migration, proliferation, and osteogenic differentiation was further investigated in the milieu of autophagy activator, rapamycin, and inhibitor, 3-methyladenine. Results showed that PRP promoted cell migration, proliferation, and osteogenic differentiation. Autophagic bodies were strongly activated and the expression level of LC3B and Beclin-1 was significantly promoted by PRP. Autophagy inhibition suppressed PRP-induced hDPCs migration, proliferation, and osteogenic differentiation, whereas autophagy activator substantially augmented PRP-stimulated migration, proliferation, and differentiation. Taken together, these findings suggested that PRP could effectively promote regenerative potentials associated with autophagy.
Highlights
If immature teeth have a diffuse pulp infection or periapical periodontitis with an open apex, apexification with calcium hydroxide or apical barrier with mineral trioxide aggregate was adapted according to the traditional methods (Bonte et al, 2015)
We explored the effects of plateletrich plasma (PRP) in the autophagic activity of Human dental pulp cells (hDPCs) and the role of autophagy in PRP-induced migration, proliferation, and differentiation
The results showed that 5, 10, and 20% PRP significantly promoted cell migration of hDPCs (Figure 1A) and 5% PRP showed the best effect
Summary
If immature teeth have a diffuse pulp infection or periapical periodontitis with an open apex, apexification with calcium hydroxide or apical barrier with mineral trioxide aggregate was adapted according to the traditional methods (Bonte et al, 2015). Neither method above could promote root development (Andreasen et al, 2002). PRP separated from whole blood has been described as platelet concentrates. When these platelets are activated, they could release amounts of growth factors. These growth factors are confirmed to accelerate the healing process and promote pulp repair (Altaii et al, 2017). PRP presented benefits as a natural scaffold for cell proliferation, migration, and differentiation in pulp regeneration (Torabinejad and Faras, 2012; Bezgin et al, 2015)
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