TY - JOUR
T1 - Effects of initial seeding density and fluid perfusion rate on formation of tissue-engineered bone
AU - Grayson, Warren L.
AU - Bhumiratana, Sarindr
AU - Cannizzaro, Christopher
AU - Chao, P. H.Grace
AU - Lennon, Donald P.
AU - Caplan, Arnold I.
AU - Vunjak-Novakovic, Gordana
PY - 2008/11/1
Y1 - 2008/11/1
N2 - We describe a novel bioreactor system for tissue engineering of bone that enables cultivation of up to six tissue constructs simultaneously, with direct perfusion and imaging capability. The bioreactor was used to investigate the relative effects of initial seeding density and medium perfusion rate on the growth and osteogenic differentiation patterns of bone marrow-derived human mesenchymal stem cells (hMSCs) cultured on three-dimensional scaffolds. Fully decellularized bovine trabecular bone was used as a scaffold because it provided suitable "biomimetic" topography, biochemical composition, and mechanical properties for osteogenic differentiation of hMSCs. Trabecular bone plugs were completely denuded of cellular material using a serial treatment with hypotonic buffers and detergents, seeded with hMSCs, and cultured for 5 weeks. Increasing seeding density from 30 × 106 cells/mL to 60 × 106 cells/mL did not measurably influence the characteristics of tissue-engineered bone, in contrast to an increase in the perfusion rate from 100 μms-1 to 400 μms-1, which radically improved final cell numbers, cell distributions throughout the constructs, and the amounts of bone proteins and minerals. Taken together, these findings suggest that the rate of medium perfusion during cultivation has a significant effect on the characteristics of engineered bone.
AB - We describe a novel bioreactor system for tissue engineering of bone that enables cultivation of up to six tissue constructs simultaneously, with direct perfusion and imaging capability. The bioreactor was used to investigate the relative effects of initial seeding density and medium perfusion rate on the growth and osteogenic differentiation patterns of bone marrow-derived human mesenchymal stem cells (hMSCs) cultured on three-dimensional scaffolds. Fully decellularized bovine trabecular bone was used as a scaffold because it provided suitable "biomimetic" topography, biochemical composition, and mechanical properties for osteogenic differentiation of hMSCs. Trabecular bone plugs were completely denuded of cellular material using a serial treatment with hypotonic buffers and detergents, seeded with hMSCs, and cultured for 5 weeks. Increasing seeding density from 30 × 106 cells/mL to 60 × 106 cells/mL did not measurably influence the characteristics of tissue-engineered bone, in contrast to an increase in the perfusion rate from 100 μms-1 to 400 μms-1, which radically improved final cell numbers, cell distributions throughout the constructs, and the amounts of bone proteins and minerals. Taken together, these findings suggest that the rate of medium perfusion during cultivation has a significant effect on the characteristics of engineered bone.
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U2 - 10.1089/ten.tea.2007.0255
DO - 10.1089/ten.tea.2007.0255
M3 - Article
C2 - 18620487
AN - SCOPUS:54949104711
SN - 1937-3341
VL - 14
SP - 1809
EP - 1820
JO - Tissue Engineering - Part A.
JF - Tissue Engineering - Part A.
IS - 11
ER -