Effects of initial seeding density and fluid perfusion rate on formation of tissue-engineered bone

Warren L. Grayson, Sarindr Bhumiratana, Christopher Cannizzaro, P. H.Grace Chao, Donald P. Lennon, Arnold I. Caplan, Gordana Vunjak-Novakovic

Research output: Contribution to journalArticlepeer-review

179 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)1809-1820
Number of pages12
JournalTissue Engineering - Part A.
Issue number11
StatePublished - Nov 1 2008
Externally publishedYes

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biomedical Engineering
  • Biomaterials


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