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

J Cell Biochem. 2001 Jun 26-Jul 25;83(1):121-8. doi: 10.1002/jcb.1203.

Abstract

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 alpha-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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actins / analysis
  • Animals
  • Bioartificial Organs*
  • Biotechnology / methods*
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins / pharmacology*
  • Cartilage / chemistry
  • Cartilage / cytology
  • Cartilage / drug effects
  • Cartilage / growth & development*
  • Cattle
  • Cell Culture Techniques / methods*
  • Cell Division / drug effects
  • Cells, Cultured
  • Chondrocytes / chemistry
  • Chondrocytes / cytology
  • Chondrocytes / drug effects*
  • Collagen / analysis
  • Fibroblast Growth Factor 2 / pharmacology*
  • Glycosaminoglycans / analysis
  • Microscopy
  • Polymers / chemistry
  • Polymers / metabolism
  • Transforming Growth Factor beta*

Substances

  • Actins
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins
  • Glycosaminoglycans
  • Polymers
  • Transforming Growth Factor beta
  • Fibroblast Growth Factor 2
  • Collagen