Cardiovascular tissue engineering: a new laminar flow chamber for in vitro improvement of mechanical tissue properties

ASAIO J. 2002 Jan-Feb;48(1):8-11. doi: 10.1097/00002480-200201000-00003.

Abstract

A new in vitro flow system was developed to investigate the impact of laminar flow on extracellular matrix formation and tissue development. The dynamic in vitro system was designed to provide a cross flow arrangement of main flow induced by a dialysis roller pump (500 ml/min), and nutrition flow by a perfusion pump (3 ml/hr). Poly-L-lysine precoated polyglycolic acid (PGA) scaffolds (3.14 cm2) were seeded with myofibroblasts of human aortic origin (3.0 x 10(6) cells/ mesh) and incubated for 14 days under static conditions. The tissue was exposed to shear stress over a time period of 14 days (n = 4). The control group was seeded under static conditions (n = 4). To obtain a CO2 independent medium, 25 mM HEPES and 1 mM bicarbonate buffer was supplemented to modified MEM without bicarbonate. Gas samples were collected from the medium, and hydroxyproline assay was performed as a marker of collagen production. The newly developed flow system maintained stable cell culture conditions, with the hydroxyproline concentration significantly higher in group F (p < 0.05). These preliminary experiences with a new in vitro tissue culture system demonstrate the feasibility of using flow induced mechanical stress to enhance extracellular matrix formation.

MeSH terms

  • Aorta / cytology*
  • Carbon Dioxide / metabolism
  • Cells, Cultured
  • Collagen / metabolism
  • Culture Media
  • Diffusion Chambers, Culture / instrumentation*
  • Extracellular Matrix / metabolism
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Humans
  • Hydroxyproline / metabolism
  • Stress, Mechanical
  • Tissue Engineering / instrumentation*
  • Tissue Engineering / methods*

Substances

  • Culture Media
  • Carbon Dioxide
  • Collagen
  • Hydroxyproline