The potential application of concentrated growth factor in pulp regeneration: an in vitro and in vivo study

Abstract

Background: Concentrated growth factor (CGF), as a natural biomaterial, is known to contain platelets, cytokines, and growth factors to facilitate the healing process, but there has been little information acquired in regenerative endodontics. The purpose of this study was to investigate the effects of CGF on proliferation, migration, and differentiation in human dental stem pulp cells (hDPSCs) exposed to lipopolysaccharide (LPS) in vitro and its potential role in pulp regeneration of the immature teeth in vivo. Methods: In vitro experiments: CGF-conditioned medium were extracted by freeze-dried method. hDPSCs were isolated and identified. The proliferative potential of hDPSCs with different concentration of CGF and LPS was evaluated by Cell Counting Kit-8. Migration capacity was analyzed by Transwell assays, odonto/osteoblastic differentiation was determined by measuring alkaline phosphatase (ALP) activity using ALP staining, and the extent of mineralization was evaluated by using Alizarin red S staining. The mRNA expression level of DMP-1, DSPP, OPN, Runx2, and OCN were determined by quantitative polymerase chain reaction (qPCR). In vivo experiments: CGF were used as root canal filling agent of the immature single-rooted teeth in the beagle dogs. The teeth were then radiographed, extracted, fixed, demineralized, and subjected to histologic analyses at 8 weeks. The newly formed dentine-pulp complex and the development of apical foramen were evaluated by the hematoxylin-eosin (HE) and Masson trichrome technique. Soft tissues were analyzed by immunohistochemical staining of vascular endothelial growth factor (VEGF) and Nestin. Results: In vitro experiments: The cultured cells exhibited the characteristics of mesenchymal stem cell. The treatment of LPS significantly increased the expression of TNF-α, IL-1β, IL-6, and IL-8 in hDPSCs, and CGF inhibited the mRNA expression of IL-8 in LPS-stimulated hDPSCs. The proliferation values of the CGF group in LPS-stimulated hDPSCs were significantly higher than that of the control group from day 3 to day 7 (P < 0.05). In addition, the number of migratory cells of the CGF group was greater than that of the control group at 24 h with or without LPS treatment. ALP activities increased gradually in both groups from day 4 to day 7. The mineralized nodules and the expression of odontogenesis-related genes DMP-1 and DSPP, osteogenesis-related genes OPN, Runx2, and OCN were dramatically enhanced by CGF in LPS-stimulated hDPSCs at days 21 and 28. In vivo experiments: In CGF treated group, the results of radiograph, HE, and Masson trichrome staining showed a continuing developed tooth of the immature teeth in the beagle dogs (i.e., the ingrowth of soft tissues into the root

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

“…With persistence of stimulus, the inflammation will cause pulp necrosis and degeneration. These processes can be terminated by appropriate treatment, and the biological structure and function of dental pulp can be partly or entirely regenerated [19]. Pulp tissue injury can be repaired by DPCs, which reside in normal pulp [3].…”

Therapeutic effect of concentrated growth factor preparation on skin photoaging in a mouse model

“…CGF is a relatively recent discovery and it has thus been the subject of limited research. However, CGF has been reported to increase the proliferation and promote the osteogenic differentiation and angiogenic potential of periosteum-derived cells, 25 promote the proliferation, migration, and differentiation of human dental stem pulp cells exposed to lipopolysaccharide, 26 and increase the proliferation and secretion of neurotrophic factors by Schwann cells in vivo . 27 Our results indicated that CGF can effectively improve skin appearance and structure, alleviate pathological changes, and promote the proliferation of skin cells, leading to a therapeutic effect on skin aging.…”

“…CGF can increase the proliferation and promote osteogenic differentiation and the angiogenic potential of periosteum-derived cells. 25 It can also promote the proliferation, migration, and differentiation of human dental stem pulp cells exposed to lipopolysaccharide, 26 and induce regeneration of the dentine–pulp complex of immature teeth in vivo . In addition, CGF can upregulate the in vivo proliferation and secretion of neurotrophic factors from Schwann cells and promote functional recovery following peripheral nerve injuries.…”

Prospects of Advanced Therapy Medicinal Products–Based Therapies in Regenerative Dentistry: Current Status, Comparison with Global Trends in Medicine, and Future Perspectives

“…Sophisticated in vitro and in vivo systems have been developed 25 . Bioengineering of various constituents of dental tissues, such as dentin 26,27 , pulp [28][29][30] , enamel 31,32 , periodontal tissues 33 , or alveolar bone 34,35 , has been achieved by the combination of scaffolds, SCs, and growth factors (TE triad) 18 . Of major interest is the regeneration of neovascularized dental pulp 36,37 , of "bio-roots" 38,39 , or even fully functional teeth in animal models 40,41 .…”