Effects of collagen membranes and bone substitute differ in periodontal ligament cell microtissues and monolayers

J Periodontol. 2022 May;93(5):697-708. doi: 10.1002/JPER.21-0225. Epub 2021 Aug 7.

Abstract

Background: Barrier membranes and bone substitute are major tools of guided tissue regeneration (GTR) after periodontal disease. Integrity of the periodontal ligament plays a key role in periodontal health, but its functionality fails to be fully re-established by GTR after disease or trauma. Microtissue models suggest an in vivo-like model to develop novel GTR approaches due to its three-dimensionality. This study aims to assess the effects of collagen membranes and bone substitute on cell viability, adhesion and gene expression of regenerative and inflammatory biomarkers by periodontal ligament cell (PDLC) microtissues.

Methods: Human PDLC microtissues and monolayers were cultured on collagen membranes or bone substitute. After 24 hours incubation, metabolic activity, focal adhesion, mRNA and protein production of collagen-type-I (COL1A1), periostin (POSTN), vascular endothelial growth factor (VEGF), angiogenin (ANG), interleukin (IL)6 and IL8 were measured by resazurin-based toxicity assay, focal adhesion staining, quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively.

Results: PDLC microtissues and monolayers were viable on collagen membranes and bone substitute, but microtissues were less metabolically active. Dominant staining of actin filaments was found in PDLC microtissues on collagen membranes. COL1A1, POSTN, VEGF, ANG and IL6 were modulated in PDLC microtissues on bone substitute, while there were no significant changes on collagen membranes. PDLC monolayers showed a different character of gene expression changes.

Conclusions: PDLC microtissues and monolayers react diversely to collagen membranes and bone substitute. Further descriptive and mechanistic tests will be required to clarify the potential of PDLC microtissues as in vivo-like model for GTR.

Keywords: biocompatible materials; bone substitutes; cell culture techniques; dentistry; guided tissue regeneration; periodontal ligament.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Substitutes* / pharmacology
  • Collagen / pharmacology
  • Collagen Type I
  • Guided Tissue Regeneration, Periodontal
  • Humans
  • Membranes, Artificial
  • Periodontal Ligament*
  • Vascular Endothelial Growth Factor A

Substances

  • Bone Substitutes
  • Collagen Type I
  • Membranes, Artificial
  • Vascular Endothelial Growth Factor A
  • Collagen