Cardiovascular tissue engineering: a new laminar flow chamber for in vitro improvement of mechanical tissue properties.
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.
Jockenhoevel
Stefan
S
Clinic for Thoracic and Cardiovascular Surgery, University Hospital Aachen, Germany.
Zund
Gregor
G
Hoerstrup
Simon P
SP
Schnell
Andrea
A
Turina
Marco
M
D001011:Aorta / Q000166:cytology*; D002245:Carbon Dioxide / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D015194:Diffusion Chambers, Culture / Q000295:instrumentation*; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D006909:Hydroxyproline / Q000378:metabolism; D013314:Stress, Mechanical; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods*
D003470:Culture Media; D002245:Carbon Dioxide; D003094:Collagen; D006909:Hydroxyproline