The effects of GDF-5 and uniaxial strain on mesenchymal stem cells in 3-D culture.
Recent endeavors in tissue engineering have attempted to identify the optimal parameters to create an artificial ligament. Both mechanical and biochemical stimulation have been used by others to independently modulate growth and differentiation, although few studies have explored their interactions. We applied previously described fabrication techniques to create a highly porous (90%-95% porosity, 212-300 microm), 3-D, bioabsorbable polymer scaffold (polycaprolactone). Scaffolds were coated with bovine collagen, and growth and differentiation factor 5 (GDF-5) was added to half of the scaffolds. Scaffolds were seeded with mesenchymal stem cells and cultured in a custom bioreactor under static or cyclic strain (10% strain, 0.33 Hz) conditions. After 48 hours, both mechanical stimulation and GDF-5 increased mRNA production of collagen I, II, and scleraxis compared to control; tenascin C production was not increased. Combining stimuli did not change gene expression; however, cellular metabolism was 1.7 times higher in scaffolds treated with both stimuli. We successfully grew a line of mesenchymal stem cells in 3-D culture, and our initial data indicate mechanical stimulation and GDF-5 influenced cellular activity and mRNA production; we did not, however, observe additive synergism with the mechanical and biological stimuli.
Department of Orthopaedic Surgery, David Geffen School of Medicine, University of California Medical Center, 10833 Le Conte Avenue, Room 16-155 CHS, Los Angeles, CA 90095, USA. efarng@mednet.ucla.edu
Eugene
E
Farng
Alfonso R
AR
Urdaneta
David
D
Barba
Sean
S
Esmende
David R
DR
McAllister
D019149:Bioreactors; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002448:Cell Adhesion; D049109:Cell Proliferation; D055428:Growth Differentiation Factor 5; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D013314:Stress, Mechanical; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D019485:Bone Morphogenetic Proteins; D055428:Growth Differentiation Factor 5; D016212:Transforming Growth Factor beta