Effects of the controlled-released TGF-beta 1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold

• Published in: Biomaterials Vol. 25; no. 18; pp. 4163 - 4173
• Main Authors: Lee, Jong Eun, Kim, Ko Eun, Kwon, Ick Chan, Ahn, Hyun Jeong, Lee, Sang-Hoon, Cho, Hyunchul, Kim, Hee Joong, Seong, Sang Chul, Lee, Myung Chul
• Format: Journal Article
• Language: English
• Published: Netherlands 01-08-2004
• Subjects: Animals; Cartilage, Articular - cytology; Cartilage, Articular - drug effects; Cartilage, Articular - physiology; Cell Division - drug effects; Cell Survival - drug effects; Cells, Cultured; Chitosan - chemistry; Chondrocytes - cytology; Chondrocytes - drug effects; Chondrocytes - physiology; Coated Materials, Biocompatible - administration & dosage; Coated Materials, Biocompatible - chemistry; Collagen - chemistry; Delayed-Action Preparations - administration & dosage; Delayed-Action Preparations - chemistry; Drug Delivery Systems - methods; Extracellular Matrix - chemistry; Glycosaminoglycans - chemistry; Materials Testing; Microspheres; Porosity; Rabbits; Surface Properties; Tissue Engineering - methods; Transforming Growth Factor beta - administration & dosage; Transforming Growth Factor beta - chemistry
• ISSN: 0142-9612
• DOI: 10.1016/j.biomaterials.2003.10.057

The objectives of this study were (1) to develop a three-dimensional collagen/chitosan/glycosaminoglycan (GAG) scaffold in combination with transforming growth factor-beta1 (TGF-beta 1)-loaded chitosan microspheres, and (2) to evaluate the effect of released TGF-beta 1 on the chondrogenic potential of rabbit chondrocytes in such scaffolds. TGF-beta 1 was loaded into chitosan microspheres using an emulsion-crosslinking method. The controlled release of TGF-beta 1, as measured by enzyme-linked immunosorbent assay (ELISA), was monitored for 7 days. The porous scaffolds containing collagen and chitosan were fabricated by using a freeze drying technique and crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide (EDC) in the presence of chondroitin sulfate (CS), as a GAG component. The TGF-beta 1 microspheres were encapsulated into the scaffold at a concentration of 10 ng TGF-beta 1/scaffold and then chondrocytes were seeded in the scaffold and incubated in vitro for 3 weeks. Both proliferation rate and glycosaminoglycan (GAG) production were significantly higher in the TGF-beta 1 microsphere-incorporated scaffolds than in the control scaffolds without microspheres. Extracellular matrix staining by Safranin O and immunohistochemistry for type II collagen were elevated in the scaffold with TGF-beta 1 microspheres. These results suggest that TGF-beta 1 microspheres when incorporated into a scaffold have the potential to enhance cartilage formation.