Effect of cell seeding concentration on the quality of tissue engineered constructs loaded with adult human articular chondrocytes.
Journal article, 2008
Many aspects of the process of in vitro differentiation of chondrocytes in three-dimensional (3D) scaffolds need to be further investigated. Chitosan scaffolds were produced by freeze-drying 3% w/v 90% DDA chitosan gels. The effect of the cell seeding concentration was evaluated by culturing human adult chondrocytes in chitosan scaffolds After the first passage, cells were seeded into chitosan scaffolds with a diameter of 8 mm. The final cell seeding concentration per cm3 of chitosan scaffold was: Group A, 3 x 10(6); Group B, 6 x 10(6); Group C, 12 x 10(6); and Group D, 25 x 10(6) cells. After 14 and 28 days in 3D culture, the constructs were assesed for collagen, glucosaminoglycans and DNA content. The mechanical properties of the constructs were determined using a dynamic oscillatory shear test. The histological aspect of the constructs was evaluated using the Bern score. The collagen and GAG concentration increased, varying the cell seeding concentration. There was a significant increase in proteoglycan and hydroxyproline production between groups C and D. The sulphated GAG content increased significantly in the group D as compared to the other groups. The mechanical properties of the different constructs increased over time, from 9.6 G'/kPa at 14 days of 3D culture to 14.6 G'/kPa at 28 days under the same culture conditions. In this study we were able to determine that concentrations of 12-25 million cells/cm2 are needed to increase the matrix production and mechanical properties of human adult chondrocytes under static conditions.
Chitosan
Humans
Articular
Cells
Scanning
Cell Culture Techniques
metabolism
Chondrocytes
cytology
Cartilage
metabolism
methods
cytology
Electron
Tissue Engineering
methods
metabolism
Microscopy
Porosity
Stress
Mechanical
Cultured
Adult
Author
Sebastian Concaro
University of Gothenburg
Elin Nicklasson
University of Gothenburg
Linda Ellowsson
Anders Lindahl
University of Gothenburg
Mats Brittberg
University of Gothenburg
Paul Gatenholm
Chalmers, Chemical and Biological Engineering, Polymer Technology
Journal of tissue engineering and regenerative medicine
1932-6254 (ISSN) 19327005 (eISSN)
Vol. 2 1 14-21Subject Categories
Surgery
Cell and Molecular Biology
Other Basic Medicine
DOI
10.1002/term.60
PubMed
18265427