Hyaluronidase inhibits TGF-β-mediated rat periodontal ligament fibroblast expression of collagen and myofibroblast markers: An in vitro exploration of periodontal tissue remodeling

Arch Oral Biol. 2024 Jul:163:105980. doi: 10.1016/j.archoralbio.2024.105980. Epub 2024 Apr 26.

Abstract

Objective: To determine the effect of hyaluronic acid (HA) degradation by hyaluronidase (HYAL) in inhibiting collagen fiber production by rat periodontal ligament cells (rPDLCs).

Design: Primary rPDLCs were isolated from the euthanized rats and used for in vitro experiments. The appropriate HYAL concentration was determined through CCK-8 testing for cytotoxicity detection and Alizarin red staining for mineralization detection. RT-qPCR and western blot assays were conducted to assess the effect of HYAL, with or without TGF-β, on generation of collagen fiber constituents and expression of actin alpha 2, smooth muscle (ACTA2) of rPDLCs.

Results: Neither cell proliferation nor mineralization were significantly affected by treatment with 4 U/mL HYAL. HYAL (4 U/mL) alone downregulated type I collagen fiber (Col1a1 and Col1a2) and Acta2 mRNA expression; however, ACTA2 and COL1 protein levels were only downregulated by HYAL treatment after TGF-β induction.

Conclusions: Treatment of rPDLCs with HYAL can inhibit TGF-β-induced collagen matrix formation and myofibroblast transformation.

Keywords: Hyaluronoglucosaminidase; Periodontal ligament; Recurrence; Stress fibers.

MeSH terms

  • Actins / metabolism
  • Animals
  • Biomarkers / metabolism
  • Blotting, Western
  • Cell Proliferation* / drug effects
  • Cells, Cultured
  • Collagen Type I / metabolism
  • Collagen* / metabolism
  • Fibroblasts* / drug effects
  • Fibroblasts* / metabolism
  • Hyaluronic Acid / pharmacology
  • Hyaluronoglucosaminidase* / pharmacology
  • In Vitro Techniques
  • Male
  • Myofibroblasts* / drug effects
  • Myofibroblasts* / metabolism
  • Periodontal Ligament* / cytology
  • Periodontal Ligament* / drug effects
  • Periodontal Ligament* / metabolism
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Real-Time Polymerase Chain Reaction
  • Transforming Growth Factor beta* / metabolism

Substances

  • Hyaluronoglucosaminidase
  • Transforming Growth Factor beta
  • Collagen
  • Hyaluronic Acid
  • Actins
  • Collagen Type I
  • Biomarkers
  • RNA, Messenger