Concentrated Growth Factor Promotes Dental Pulp Cells Proliferation and Mineralization and Facilitates Recovery of Dental Pulp Tissue

Abstract

Background: Dental pulp cells (DPCs) play vital roles in the recovery of dental pulp tissue. Concentrated growth factor (CGF) can promote proliferation and mineralization of various cells. However, the functions of CGF on DPCs and dental pulp tissue are unclear. The object of our study was to identify the roles of CGF in DPCs proliferation and mineralization in vitro and to assess the effects of CGF on direct pulp capping in vivo. Material/Methods: We performed CCK-8 and Transwell assay to detect proliferation and migration activity of DPCs. Alizarin Red staining was performed to examine mineralized nodules. Alkaline phosphatase activity test was used to measure the mineralization capacity of DPCs. We assessed the odontogenic differentiation gene expression level by Western blot and qPCR. The effect of CGF on direct pulp capping in vivo were evaluated by radiography and histopathology. Results: CGF increased the number of proliferative and migratory DPCs. CGF enhanced DPCs mineralized nodules and improved the gene expression levels of DSPP, DMP-1, BSP, and ALP. CGF upregulated the protein levels of ALP, BMP2, SMAD5, Runx2, and p-Smad, and the effect could be partially reversed by Noggin. CGF promoted pulp recovery and kept its vitality in directly pulp capping. Conclusions: CGF promotes DPCs proliferation and mineralization. It regulates the mineralization of DPCs via the BMP2/SMAD5/Runx2 signaling pathway. CGF can be used as the effective graft for direct pulp capping.

Influence of Adiponectin and Leptin on the Kinetics of Human Pulp Cells

“…Stimulation of the cells with either adiponectin or leptin in a time-dependent manner increased the proliferation index of the cells as measured by BrdU incorporation, and both proteins show the same effects on human osteoblast cell proliferation [23] [24]. Conversely, stimulated pulp cells showed no change in ALP activity compared to control cells and showed no expression of Runx2 [20] [25] [26]. ALP activity and the expression of Runx2 were promoted by activated osteoblasts, so isolated pulp cells in this study might not be induced to differentiate into the calcification cells [19].…”

“…Passive calcification and fibril hyperplasia are usually observed among the aging in pulp tissue. However, these changes are found not only in teeth affected by occlusion, mastication, caries, or hypersensitivity, but also in unerupted teeth, and it is inferred that pulp cells are affected under these conditions [18] [19] [20]. Additionally, osteoblasts and osteoclasts are also regulated by adipokines, which promote bone metabolism [10] [12] [21].…”

The effects and potential applications of concentrated growth factor in dentin–pulp complex regeneration

“… The secretion of TNF-α and IL-8 in DPSCs treated by CGF could promote the DPSCs migration. Tian et al (2019) [ 36 ] h-DPSCs Proliferation, migration, and odonto/osteogenic differentiation Cell counting kit-8; Transwell assays; ALP activity, ARS, and qPCR (DMP-1, DSPP, BSP, RUNX2) CGF promoted the proliferation and migration of DPSCs in a dose-dependent manner, and CGF enhanced DPSCs odonto/osteogenic differentiation by upregulating RUNX2 transcription. BMP-2/SMAD5/Runx2 signaling axis is related to CGF-mediated DPSCs mineralization.…”

“…CGF is known to stimulate the proliferation of various MSC types (e.g., PDLSCs, DPSCs, and MSCs [hTERT-E6/E7]) in a dose-dependent manner, possibly through the independent or synergistic effects of GFs [ 36 , 37 , 40 , 42 ]. However, some studies have reported a lack of dose dependence, which may be attributable to the different methods used to prepare CGF [ 34 , 38 ].…”

“…GFs are known to act synergistically and their mechanisms of action involve the activation of Runt-related homeobox (RUNX)2, the main regulatory transcription factor in osteogenic/odontoblastic differentiation [ 59 ]. BMSCs and DPSCs cultured in CGF overexpress RUNX2 [ 36 , 41 ]. BMP-2 promotes the expression of RUNX2 via the BMP-2/Mothers against decapentaplegic homolog (SMAD)5/Runx2 signalling axis in bone formation and remodelling, which is also involved in CGF-mediated DPSC mineralisation [ 36 , 60 ].…”