iRoot BP Plus promotes osteo/odontogenic differentiation of bone marrow mesenchymal stem cells via MAPK pathways and autophagy

Stem Cell Res Ther. 2019 Jul 29;10(1):222. doi: 10.1186/s13287-019-1345-3.

Abstract

Background: iRoot BP Plus is a novel bioceramic endodontic material. Recently, it has been considered as an alternative to MTA which is the most popular scaffold cover during regenerative endodontic therapy. This study aimed to evaluate the effects of iRoot BP Plus on the osteo/odontogenic capacity of bone marrow mesenchymal stem cells (BMMSCs), including the underlying mechanisms.

Methods: BMMSCs were collected by a whole marrow method and treated with iRoot BP Plus-conditioned medium (BP-CM). The proliferation ability was evaluated by cell counting kit 8 and flow cytometry. Complete medium was used as a blank control, and 2 mg/ml MTA-conditioned medium was served as a positive control. Alkaline phosphatase (ALP) activity assay, ALP staining, western blot, real-time RT-PCR, Alizarin Red S staining, and immunofluorescence staining were performed to explore the osteo/odontogenic potential and the involvement of MAPK pathways. Besides, autophagy was investigated by western blot, immunofluorescence staining, and transmission electron microscopy.

Keywords: Autophagy; BMMSCs; MAPK pathways; Osteo/odontogenic differentiation; iRoot BP Plus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy / drug effects*
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Culture Media, Conditioned / chemistry
  • Culture Media, Conditioned / pharmacology
  • G1 Phase Cell Cycle Checkpoints / drug effects
  • MAP Kinase Signaling System / drug effects*
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Odontogenesis / drug effects*
  • Osteogenesis / drug effects*
  • Rats
  • Rats, Sprague-Dawley
  • Sequestosome-1 Protein / genetics
  • Sequestosome-1 Protein / metabolism
  • Silicates / chemistry
  • Silicates / pharmacology*
  • Sp7 Transcription Factor / genetics
  • Sp7 Transcription Factor / metabolism

Substances

  • Core Binding Factor Alpha 1 Subunit
  • Culture Media, Conditioned
  • Runx2 protein, rat
  • Sequestosome-1 Protein
  • Silicates
  • Sp7 Transcription Factor
  • Sqstm1 protein, rat
  • iRoot BP Plus