Chondrogenesis of aged human articular cartilage in a scaffold-free bioreactor.
Chondrogenesis of aged human articular chondrocytes was evaluated under controlled in vitro conditions, using a rotating bioreactor vessel. Articular chondrocytes isolated from 10 aged patients (median age, 84 years) were increased in monolayer culture. A single-cell suspension of dedifferentiated chondrocytes was inoculated in a rotating wall vessel, without the use of any scaffold or supporting gel material. After 90 days of cultivation, a three-dimensional cartilage-like tissue was formed, encapsulated by fibrous tissue resembling a perichondrial membrane. Morphological examination revealed differentiated chondrocytes ordered in clusters within a continuous dense cartilaginous matrix demonstrating a strong positive staining with monoclonal antibodies against collagen type II and articular proteoglycan. The surrounding fibrous membrane consisted of fibroblast-like cells, and showed a clear distinction from the cartilaginous areas when stained against collagen type I. Transmission electron microscopy revealed differentiated and highly metabolically active chondrocytes, producing an extracellular matrix consisting of a fine network of randomly distributed cross-banded collagen fibrils. Chondrogenesis of aged human articular chondrocytes can be induced in vitro in a rotating bioreactor vessel using low shear and efficient mass transfer. Moreover, the tissue-engineered constructs may be used for further in vitro studies of differentiation, aging, and regeneration of human articular cartilage.
Marlovits
Stefan
S
Department of Traumatology, University of Vienna Medical School, Vienna, Austria. stefan.malovits@akh-wien.ac.at
Tichy
Brigitte
B
Truppe
Michaela
M
Gruber
Daniela
D
Vécsei
Vilmos
V
D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology*; D019149:Bioreactors; D002358:Cartilage, Articular / Q000166:cytology* / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000502:physiology*; D020219:Chondrogenesis / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D006801:Humans; D023822:Tissue Engineering / Q000379:methods*