Comparison of Chondrogensis in Static and Perfused Bioreactor Culture

• Published in: Biotechnology progress Vol. 16; no. 5; pp. 893 - 896
• Main Authors: Pazzano, David, Mercier, Kathi A., Moran, John M., Fong, Stephen S., DiBiasio, David D., Rulfs, Jill X., Kohles, Sean S., Bonassar, Lawrence J.
• Format: Journal Article
• Language: English
• Published: USA American Chemical Society 01.09.2000; American Institute of Chemical Engineers
• Subjects: Animal cells; Biochemical engineering; Biological and medical sciences; Biomedical engineering; Bioreactors; Biotechnology; Cartilage; Cell Culture Techniques; Chondrocytes - cytology; Cytology; Equipment Design; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Humans; Hydrogen-Ion Concentration; Metabolism; Methods. Procedures. Technologies; pH effects; Physiology; Statistical methods; Cartilage; Cell culture; Bioreactor; Static conditions; Chondrocyte; Perfusion; Tissue engineering; Animal; Chondrogenesis; Medical application; Comparative study
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1021/bp000082v

As a result of the low yield of cartilage from primary patient harvests and a high demand for autologous cartilage for reconstructive surgery and structural repair, primary explant cartilage must be augmented by tissue engineering techniques. In this study, chondrocytes seeded on PLLA/PGA scaffolds in static culture and a direct perfusion bioreactor were biochemically and histologically analyzed to determine the effects of fluid flow and media pH on matrix assembly. A gradual media pH change was maintained in the bioreactor within 7.4−6.96 over 2 weeks compared to a more rapid decrease from 7.4 to 6.58 in static culture over 3 days. Seeded scaffolds subjected to 1 μm/s flow demonstrated a 118% increase (p < 0.05) in DNA content, a 184% increase (p < 0.05) in GAG content, and a 155% (p< 0.05) increase in hydroxyproline content compared to static culture. Distinct differences were noted in tissue morphology, including more intense staining for proteoglycans by safranin‐O and alignment of cells in the direction of media flow. Culture of chondrocyte seeded matrices thus offers the possibility of rapid in vitro expansion of donor cartilage for the repair of structural defects, tracheal injury, and vascularized tissue damage.