Osteoblast responses one hour after load-induced fluid flow in a three-dimensional porous matrix

Shigeo M. Tanaka, Hui (Herb) Sun, Ryan K. Roeder, David B. Burr, Charles H. Turner, Hiroki Yokota

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

When bone is loaded, substrate strain is generated by the external force and this strain induces fluid flow that creates fluid shear stress on bone cells. Our current understanding of load-driven gene regulation of osteoblasts is based primarily on in vitro studies on planer two-dimensional tissue culture substrates. However, differences between a flat layer of cells and cells in 3-dimensional (3D) ECM are being recognized for signal transduction. Proliferation and differentiation of osteoblasts are affected by substrate geometry. Here we developed a novel 3D culture system that would mimic physiologically relevant substrate strain as well as strain-induced fluid flow in a 3D porous collagen matrix. The system allowed us to evaluate the responses of osteoblasts in a 3D stress-strain environment similar to a mechanical field to which bone is exposed. Using MC3T3-E1 osteoblasts grown in the 3D collagen matrix with and without hydroxyapatite deposition, we tested the role of strain and the strain-induced fluid flow in the expression of the load-responsive genes such as c-fos, egr1, cox2, osteopontin, and mmp1B involved in transcriptional regulation, osteogenesis, and rearrangement of ECM. Strain-induced fluid flow was visualized with a microspheres ~3 μm in diameter in real time, and three viscoelastic parameters were determined. The results obtained by semi-quantitative PCR, immunoblot assay, enzymatic activity assays for collagenase and gelatinase, and mechanical characterization of collagen matrices supported the dominant role of strain-induced fluid flow in expression of the selected genes one hour after the mechanical treatment.

Original languageEnglish (US)
Pages (from-to)261-271
Number of pages11
JournalCalcified Tissue International
Volume76
Issue number4
DOIs
StatePublished - Apr 2005
Externally publishedYes

Fingerprint

Osteoblasts
Collagen
Bone and Bones
Gelatinases
Osteopontin
Enzyme Assays
Collagenases
Durapatite
Microspheres
Osteogenesis
Genes
Signal Transduction
Gene Expression
Polymerase Chain Reaction

Keywords

  • Bone
  • Collagen matrix
  • Fluid flow shear
  • Hydroxyapatite
  • Mechanical loading
  • Osteoblasts

ASJC Scopus subject areas

  • Endocrinology

Cite this

Osteoblast responses one hour after load-induced fluid flow in a three-dimensional porous matrix. / Tanaka, Shigeo M.; Sun, Hui (Herb); Roeder, Ryan K.; Burr, David B.; Turner, Charles H.; Yokota, Hiroki.

In: Calcified Tissue International, Vol. 76, No. 4, 04.2005, p. 261-271.

Research output: Contribution to journalArticle

Tanaka, Shigeo M. ; Sun, Hui (Herb) ; Roeder, Ryan K. ; Burr, David B. ; Turner, Charles H. ; Yokota, Hiroki. / Osteoblast responses one hour after load-induced fluid flow in a three-dimensional porous matrix. In: Calcified Tissue International. 2005 ; Vol. 76, No. 4. pp. 261-271.
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