The Potential Application of Human Gingival Fibroblast-Conditioned Media in Pulp Regeneration: An In Vitro Study

Cells. 2022 Oct 27;11(21):3398. doi: 10.3390/cells11213398.

Abstract

Regenerative endodontic treatment based on tissue engineering has recently gained interest in contemporary restorative dentistry. However, low survival rates and poor potential differentiation of stem cells could undermine the success rate of pulp regenerative therapy. Human gingival fibroblast-conditioned medium (hGF-CM) has been considered a potential therapy for tissue regeneration due to its stability in maintaining multiple factors essential for tissue regeneration compared to live cell transplantation. This study aimed to investigate the potency of hGF-CM on stem cells from human dental pulp (DPSC) in pulp regeneration. A series of experiments confirmed that hGF-CM contributes to a significant increase in proliferation, migration capability, and cell viability of DPSC after H2O2 exposure. Moreover, it has been proved to facilitate the odontogenic differentiation of DPSC via qRT-PCR, ALP (alkaline phosphatase), and ARS (Alizarin Red S) staining. It has been discovered that such highly upregulated odontogenesis is related to certain types of ECM proteins (collagen and laminin) from hGF-CM via proteomics. In addition, it is found that the ERK pathway is a key mechanism via inhibition assay based on RNA-seq result. These findings demonstrate that hGF-CM could be beneficial biomolecules for pulp regeneration.

Keywords: conditioned medium; gingival fibroblast; human exfoliated deciduous teeth; odontoblast differentiation; pulp regeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Culture Media, Conditioned* / chemistry
  • Culture Media, Conditioned* / pharmacology
  • Dental Pulp* / drug effects
  • Dental Pulp* / metabolism
  • Fibroblasts / metabolism
  • Gingiva / cytology
  • Gingiva / metabolism
  • Humans
  • Hydrogen Peroxide*
  • Regeneration
  • Tissue Engineering* / methods

Substances

  • Alkaline Phosphatase
  • Culture Media, Conditioned
  • Hydrogen Peroxide

Grants and funding

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Ministry of Education or the Ministry of Science and Technology (2019R1A6A1A11034536, 2018K1A4A3A01064257, 2021R1A5A2022318, 2022R1F1A1063017, and 2020R1G1A1009155, 2022K1A3A1A08085419). This research was also supported by the Department of Dentistry (Pediatric Dentistry) through the Research-Focused Department Promotion Project as a part of the University Innovation Support Program for Dankook University in 2021 and the Priority Institute Support Program (Basic Dental Scientist Support Program) in 2021. In addition, the work was partly supported by the Starting growth Technological R&D Program (S3282274) funded by the Small and Medium Business Administration (SMBA, Korea).