Chondrogenic differentiation of human bone marrow mesenchymal stem cells in chitosan-based scaffolds using a flow-perfusion bioreactor

J Tissue Eng Regen Med. 2011 Oct;5(9):722-32. doi: 10.1002/term.372. Epub 2010 Dec 29.

Abstract

Native articular cartilage is subjected to synovial fluid flow during normal joint function. Thus, it is believed that the morphogenesis of articular cartilage may be positively regulated by the application of similar stimulation in vitro. In the present study, the effect of fluid flow over the chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) was investigated. We intended to find out whether the shear stress caused by perfusion of the medium through the constructs was capable of augmenting the differentiation process. Human BMSCs were isolated from bone marrow aspirates and were characterized by flow cytometry. After expansion, hBM-MSCs were seeded statically onto fibre mesh scaffolds, consisting of a blend of 50:50 chitosan:poly(butylene terephthalate adipate) (CPBTA). Constructs were cultured in a flow-perfusion bioreactor for 28 days, using complete medium for chondrogenesis supplemented by TGFβ3. An enhanced ECM deposition and collagen type II production was observed in the bioreactor samples when compared to the static controls. Moreover, it was observed that hBM-MSCs, in static cultures, take longer to differentiate. ECM accumulation in these samples is lower than in the bioreactor sections, and there is a significant difference in the expression of collagen type I. We found that the flow-induced shear stress has a beneficial effect on the chondrogenic differentiation of hMSCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism
  • Bioreactors*
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Bone Marrow Cells / ultrastructure
  • Cell Differentiation / drug effects*
  • Cell Separation
  • Cell Shape / drug effects
  • Cells, Cultured
  • Chitosan / pharmacology*
  • Chondrogenesis / drug effects*
  • Chondrogenesis / genetics
  • Collagen Type I / metabolism
  • Collagen Type II / metabolism
  • DNA / metabolism
  • Flow Cytometry
  • Humans
  • Immunohistochemistry
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / metabolism
  • Mesenchymal Stem Cells / ultrastructure
  • Perfusion
  • Polyesters / pharmacology
  • Real-Time Polymerase Chain Reaction
  • Rheology / drug effects
  • Staining and Labeling
  • Tissue Engineering / instrumentation*
  • Tissue Scaffolds / chemistry

Substances

  • Biomarkers
  • Collagen Type I
  • Collagen Type II
  • Polyesters
  • poly(butylene adipate-co-butylene terephthalate)
  • DNA
  • Chitosan