Chondrogenesis on BMP-6 loaded chitosan scaffolds in stationary and dynamic cultures

Biotechnol Bioeng. 2009 Oct 15;104(3):601-10. doi: 10.1002/bit.22426.

Abstract

We originally investigated the suitability of chitosan scaffolds loaded with bone morphogenetic protein 6 (BMP-6) in both stationary and dynamic conditions for cartilage tissue engineering. In the first part of the present study, ATDC5 murine chondrogenic cells were seeded in chitosan and BMP-6 loaded chitosan scaffolds and cultured for 28 days under static conditions. In the following part, we examined the influence of dynamic cultivation conditions over BMP-6 loaded chitosan scaffolds by using rotating bioreactor with perfusion (RCMW). Tissue engineered constructs were characterized by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT) assay, scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and biochemical assays for glycosaminoglycans (GAG) deoxyribonucleic acid (DNA) and collagen Type II quantification. At the end of 4 weeks static incubation period high levels of GAG (21.22 mg/g dry weight), DNA amounts (1.37 mg/g dry weight) and collagen Type II amounts (1.94 microg/g dry weight) were achieved for BMP-6 loaded chitosan scaffolds compared to chitosan scaffolds. However, the results obtained from morphological observations suggested hypertrophic differentiation of ATDC5 cells in the presence of BMP-6 under stationary conditions. The influence of mechanical stimulation appeared significantly with differentiated cells, cultured under dynamic conditions, showing the effect of retaining their phenotypes without hypertrophy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 6 / metabolism*
  • Cell Survival
  • Chitosan*
  • Chondrocytes / drug effects*
  • Chondrocytes / ultrastructure
  • Chondrogenesis*
  • Collagen Type II / analysis
  • DNA / analysis
  • Glycosaminoglycans / analysis
  • Mice
  • Microscopy, Confocal
  • Microscopy, Electron, Scanning
  • Tetrazolium Salts / metabolism
  • Thiazoles / metabolism
  • Tissue Engineering / methods*

Substances

  • Bone Morphogenetic Protein 6
  • Collagen Type II
  • Glycosaminoglycans
  • Tetrazolium Salts
  • Thiazoles
  • DNA
  • Chitosan
  • thiazolyl blue