Human chondrocyte proliferation and matrix synthesis cultured in Atelocollagen� gel

Abstract

To evaluate the potential of Atelocollagen gel as a carrier for chondrocyte transplantation, histological and biochemical characteristics of the chondrocytes in gel culture were compared with those in conventional monolayer cultures. Articular chondrocytes from 20 patients were isolated by enzyme digestion, embedded in Atelocollagen gel, and cultured for up to 4 weeks. The effects on proliferation, morphological changes, and synthesis of proteoglycans were analyzed by cell counts, light and electron microscopy, and measurement of isomers of chondroitin sulfates. Chondrocytes embedded in the Atelocollagen gel gradually proliferated and produced chondroitin 6-sulfate, maintaining the chondrocyte phenotype for up to 4 weeks. In contrast, although monolayer chondrocytes increased in number, most could be characterized as being fibroblast-like cells with a reduced capability of producing chondroitin 6-sulfate. The results suggest that Atelocollagen gel permitted a gradual proliferation and matrix synthesis of chondrocytes and maintaining its phenotype. Atelocollagen gel represents an important carrier for the clinical application of cultured chondrocyte transplantation for repair of cartilage defects.

Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

“…Interaction of this subunit with collagen type I leads to osteoblastic differentiation of MSCs, which is a crucial event in the expression of the osteogenic phenotype (Mizuno et al, 2000). Chondrocytes cultured in collagen type I gel grew in three dimensions, accumulating a cartilaginous ECM while maintaining their round morphology during the culture period (Kimura et al, 1987;Uchio et al, 2002). Furthermore, a cartilage defect was partially repaired when human autologous culture-expanded MSCs engineered in a collagen type I gel were transplanted into an osteoarthritic knee (Wakitani et al, 2002).…”

Transplantation of cartilage-like tissue made by tissue engineering in the treatment of cartilage defects of the knee

“…6 Our previous in vitro study in which human articular chondrocytes were used also showed that chondrocytes in a monolayer culture differentiated into fibroblast-like cells and synthesised chondroitin-4-sulphatedominant proteoglycans. 25 By contrast, chondrocytes cultured in atelocollagen gel maintained the chondrocyte phenotype and synthesised a chondroitin-6-sulphate-rich matrix. In addition, Coon and Cahn 35 reported that dedifferentiated chondrocytes, during four successive clonal passages in monolayer culture, re-expressed chondrocyte phenotypes when they were in contact with normal chondrocytes or were in the presence of unknown highmolecular-weight molecules.…”

Tissue Engineering von Knorpelersatzgewebe – Mechanische Stimulation in der In-vitro-Kultivierung von humanen Chondrozyten

“…Auch als dreidimensionales Kultivierungssystem haben Kollagenmatrices im Rahmen von MACT-Verfahren einen klinischen Stellenwert (Andereya et al, 2006;Andereya et al, 2007). Kollagengelmatrices stimulieren die zelluläre Proliferation und Syntheseleistung in vitro (Schuman et al, 1995;Uchio et al, 2000) und stellen damit zumindest in vielerlei Hinsicht eine funktionelle Extrazellulär-matrix-Umgebung dar, die hyalinem Knorpel ähnlich ist (Schneider et al, 2003) und…”