Cell survival and gene expression under compressive stress in a three-dimensional in vitro human periodontal ligament-like tissue model

Wen Liao, Masahiro Okada, Kaoru Inami, Yoshiya Hashimoto, Naoyuki Matsumoto

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

This study investigated cell survival and gene expression under various compressive stress conditions mimicking orthodontic force by using a newly developed in vitro model of human periodontal ligament-like tissue (HPdLLT). The HPdLLT was developed by three-dimensional culturing of human periodontal ligament fibroblasts in a porous poly-l-lactide matrix with threefold increased culture media permeability due to hydrophilic modification. In vitro HPdLLTs in experimental groups were subjected to 5, 15, 25 and 35 g/cm2 compressive stress for 1, 3, 7 or 14 days; controls were cultured over the same periods without compressive stress. Cell morphology and cell apoptosis in the experimental and control groups were investigated using scanning electron microscopy and caspase-3/7 detection. Real-time polymerase chain reaction was performed for seven osteogenic and osteoclastic genes. Similar extracellular matrix and spindle-shaped cells were observed inside or on the surface of in vitro HPdLLTs, with no relation to compressive stress duration or intensity. Similar caspase-3/7 activity indicating comparable apoptosis levels was observed in all samples. Receptor activator of nuclear factor kappa-B ligand and bone morphogenetic protein 2 genes showed characteristic “double-peak” expression at 15 and 35 g/cm2 on day 14, and alkaline phosphatase and periodontal ligament-associated protein 1 expression peaked at 5 g/cm2 on day 14; other genes also showed time-dependent and load-dependent expression patterns. The in vitro HPdLLT model system effectively mimicked the reaction and gene expression of the human periodontal ligament in response to orthodontic force. This work provides new information on the effects of compressive stress on human periodontal ligament tissue.

Original languageEnglish
Pages (from-to)249-260
Number of pages12
JournalCytotechnology
Volume68
Issue number2
DOIs
Publication statusPublished - Mar 1 2016
Externally publishedYes

Keywords

  • Cell living condition
  • Compressive stress
  • Gene expression
  • Human periodontal ligament-like tissue
  • Orthodontic force

ASJC Scopus subject areas

  • Cell Biology
  • Biotechnology
  • Biomedical Engineering
  • Bioengineering
  • Clinical Biochemistry

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