Collagen Expression in Tissue Engineered Cartilage of Aged Human Articular Chondrocytes in a Rotating Bioreactor

Abstract

This study describes the culture and three-dimensional assembly of aged human articular chondrocytes under controlled oxygenation and low shear stress in a rotating-wall vessel. Chondrocytes cultured in monolayer were released and placed without any scaffold as a single cell suspension in a rotating bioreactor for 12 weeks. Samples were analyzed with immunohistochemistry, molecular biology and electron microscopy. During serial monolayer cultures chondrocytes dedifferentiated to a “fibroblast-like” structure and produced predominantly collagen type I. When these dedifferentiated cells were transferred to the rotating bioreactor, the cells showed a spontaneous aggregation and formation of solid tissue during the culture time. Expression of collagen type II and other components critical for the extracellular cartilage matrix could be detected. Transmission electron microscopy revealed a fine network of randomly distributed collagen fibrils. This rotating bioreactor proves to be a useful tool for providing an environment that enables dedifferentiated chondrocytes to redifferentiate and produce a cartilage-specific extracellular matrix.

Chondrogenic differentiation of human adipose-derived stem cells using microcarrier and bioreactor combination technique

“…It is difficult to meet these requirements with traditional static culture, whilst three-dimensional dynamic culture is able to accelerate cell proliferation and be conducive to chondrogenic differentiation and phenotype maintenance (7). Therefore, the present study applied bioreactor technology combined with microcarriers to rapidly amplify and induce chondrogenic differentiation in ADSCs.…”

Neocartilage Formation in 1 g, Simulated, and Microgravity Environments: Implications for Tissue Engineering

“…[13][14][15][16][17] More recently, Ohyabu et al 18 used rabbit bone marrow stromal cells in an RWV to produce cartilage tissue. Conza et al 19 showed that neocartilage cultivated on an RPM exhibits a much more regular distribution of cells within a well-developed cartilage extracellular matrix (ECM) than the layered structures produced under normal conditions without a supporting scaffold.…”

Comparison of articular and auricular cartilage as a cell source for the autologous chondrocyte implantation

“…Appropriate bioreactor and mechanical stimuli may further improve the chondrogenic phenotype. 29 A similar 3D hydrogel, alginate-agarose, seeded with autologous articular chondrocytes, has recently been introduced into clinical work; excellent clinical, radiological, and histological results in the treatment of femoral condyle osteochondral lesions were demonstrated. 30 Relatively high ELS in the auricular chondrocytes revealed their capability to retain some characteristics of auricular cartilage tissue after 1 week in vitro culturing.…”