Low-energy red light-emitting diode irradiation enhances osteogenic differentiation of periodontal ligament stem cells by regulating miR-146a-5p

J Periodontal Res. 2024 Oct;59(5):1031-1041. doi: 10.1111/jre.13276. Epub 2024 Jun 6.

Abstract

Aims: The study aimed to investigate the role of miR-146a-5p in osteogenesis of hPDLSCs irradiated with low-energy red LEDs.

Methods: After irradiation with 5 J/cm2 red LED, miR-146a-5p expression was detected by real-time quantitative polymerase chain reaction (RT-qPCR), and osteogenic markers expression was determined by RT-qPCR and Western blotting. Alkaline phosphatase (ALP) activity was assessed by ALP staining, and mineralization was assessed by Alizarin Red staining, respectively. Lentiviral vectors were designed to regulate miR-146a-5p expression. Dual-luciferase reporter assay was performed to confirm the targeted relationship between miR-146a-5p and MAPK1. Short hairpin RNA (shRNA) was used to regulate MAPK1 expression.

Results: RT-qPCR and western blotting revealed that 5 J/cm2 irradiation elevated the levels of the osteogenic markers osterix (OSX) and bone sialoprotein (BSP) in hPDLSCs. miR-146a-5p is downregulated in hPDLSCs under the low-energy red LED light irradiation. miR-146a-5p underexpression markedly promoted the osteogenic potential of hPDLSCs. miR-146a-5p targeted MAPK1. 5 J/cm2 red LED irradiation rescued the inhibitory effects of upregulated miR-146a-5p on osteogenic differentiation, and the positive influence of red LED irradiation could be reversed by downregulated MAPK1.

Conclusion: These findings confirm that miR-146a-5p is involved in the effect of LED irradiation on the osteogenic differentiation of hPDLSCs by targeting MAPK1. Red LED irradiation may be a potential clinical adjunct therapy for periodontal regeneration.

Keywords: LED; hPDLSCs; miR‐146a‐5p; osteogenic differentiation.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Cell Differentiation* / radiation effects
  • Cells, Cultured
  • Humans
  • Integrin-Binding Sialoprotein
  • Light
  • MicroRNAs*
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Osteogenesis* / radiation effects
  • Periodontal Ligament* / cytology
  • Periodontal Ligament* / radiation effects
  • Real-Time Polymerase Chain Reaction
  • Red Light
  • Sp7 Transcription Factor
  • Stem Cells* / radiation effects

Substances

  • MicroRNAs
  • MIRN146 microRNA, human
  • Integrin-Binding Sialoprotein
  • Alkaline Phosphatase
  • Mitogen-Activated Protein Kinase 1
  • MAPK1 protein, human
  • Sp7 Transcription Factor