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Enhanced cartilage tissue engineering by sequential exposure of chondrocytes to FGF‐2 during 2D expansion and BMP‐2 during 3D cultivation

Bovine calf articular chondrocytes, either primary or expanded in monolayers (2D) with or without 5 ng/ml fibroblast growth factor‐2 (FGF‐2), were cultured on three‐dimensional (3D) biodegradable polyglycolic acid (PGA) scaffolds with or without 10 ng/ml bone morphogenetic protein‐2 (BMP‐2). Chondrocytes expanded without FGF‐2 exhibited high intensity immunostaining for smooth muscle α‐actin (SMA) and collagen type I and induced shrinkage of the PGA scaffold, thus resembling contractile fibroblasts. Chondrocytes expanded in the presence of FGF‐2 and cultured 6 weeks on PGA scaffolds yielded engineered cartilage with 3.7‐fold higher cell number, 4.2‐fold higher wet weight, and 2.8‐fold higher wet weight glycosaminoglycan (GAG) fraction than chondrocytes expanded without FGF‐2. Chondrocytes expanded with FGF‐2 and cultured on PGA scaffolds in the presence of BMP‐2 for 6 weeks yielded engineered cartilage with similar cellularity and size, 1.5‐fold higher wet weight GAG fraction, and more homogenous GAG distribution than the corresponding engineered cartilage cultured without BMP‐2. The presence of BMP‐2 during 3D culture had no apparent effect on primary chondrocytes or those expanded without FGF‐2. In summary, the presence of FGF‐2 during 2D expansion reduced chondrocyte expression of fibroblastic molecules and induced responsiveness to BMP‐2 during 3D cultivation on PGA scaffolds. © 2001 Wiley‐Liss, Inc.