Biologic interaction of three-dimensional periodontal fibroblast spheroids with collagen-based and synthetic membranes

J Periodontol. 2011 May;82(5):790-7. doi: 10.1902/jop.2010.100533. Epub 2010 Nov 16.

Abstract

Background: Cell-based therapy using autologous cells has been suggested as a potential approach for periodontal tissue regeneration. Spheroid systems are a form of three-dimensional cell culture that promotes cell matrix interaction, which could recapitulate the aspect of cell homeostasis in vivo. The aim of this study is to assess the interaction of periodontal fibroblast spheroids with synthetic and collagen-based membranes that have been used in guided tissue regeneration.

Methods: Commercially available normal human periodontal ligament fibroblasts were grown in spheroid forms using a liquid overlay technique and then transplanted onto a collagen-based and a polyglycolic acid-based membrane. The biologic interaction of the spheroids with the membranes was assessed using basic histology, Alamar blue tissue viability assay, scanning electron microscopy, and immunohistochemical analysis.

Results: Periodontal fibroblast spheroids adhered to both membranes, and the cells were able to proliferate and migrate from the spheroids both horizontally and vertically into the membrane scaffolds. Immunohistochemical analysis showed expression of collagen type I, periostin, and Runx2 by the periodontal fibroblasts.

Conclusion: Periodontal fibroblast spheroids were able to grow three-dimensionally on the biologic membranes and may have the potential to be used together with guided tissue regeneration approaches as an adjunct for periodontal regeneration.

MeSH terms

  • Biocompatible Materials / chemistry*
  • Cell Adhesion / physiology
  • Cell Adhesion Molecules / analysis
  • Cell Culture Techniques
  • Cell Movement / physiology
  • Cell Proliferation
  • Cell Survival / physiology
  • Collagen / chemistry*
  • Collagen Type I / analysis
  • Core Binding Factor Alpha 1 Subunit / analysis
  • Fibroblasts / physiology*
  • Guided Tissue Regeneration, Periodontal / instrumentation
  • Humans
  • Immunohistochemistry
  • Indicators and Reagents
  • Membranes, Artificial*
  • Microscopy, Electron, Scanning
  • Oxazines
  • Periodontal Ligament / cytology*
  • Polyglycolic Acid / chemistry
  • Spheroids, Cellular / physiology*
  • Tissue Scaffolds*
  • Xanthenes

Substances

  • Biocompatible Materials
  • Cell Adhesion Molecules
  • Collagen Type I
  • Core Binding Factor Alpha 1 Subunit
  • Indicators and Reagents
  • Membranes, Artificial
  • Oxazines
  • POSTN protein, human
  • RUNX2 protein, human
  • Xanthenes
  • resazurin
  • Polyglycolic Acid
  • Collagen