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Effects of a Chitosan Scaffold Containing TGF‐β1 Encapsulated Chitosan Microspheres on In Vitro Chondrocyte Culture
Author(s) -
Lee Jong Eun,
Kim Seoung Eun,
Kwon Ick Chan,
Ahn Hyun Jeong,
Cho Hyunchul,
Lee SangHoon,
Kim Hee Joong,
Seong Sang Cheol,
Lee Myung Chul
Publication year - 2004
Publication title -
artificial organs
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.684
H-Index - 76
eISSN - 1525-1594
pISSN - 0160-564X
DOI - 10.1111/j.1525-1594.2004.00020.x
Subject(s) - chitosan , scaffold , chondrocyte , chemistry , extracellular matrix , glycosaminoglycan , chondrogenesis , in vitro , biomedical engineering , tissue engineering , in vivo , transforming growth factor , biophysics , materials science , biochemistry , microbiology and biotechnology , medicine , biology
The objectives of this study were (1) to develop a three‐dimensional chitosan scaffold in combination with transforming growth factor‐beta1 (TGF‐β1)‐loaded chitosan microspheres and (2) to evaluate the effect of the TGF‐β1 release on the chondrogenic potential of rabbit chondrocytes in the scaffolds. TGF‐β1 was loaded into chitosan microspheres using an emulsion‐crosslinking method, resulting in spherical shapes with a size ranging from 0.3 to 1.5 µm. Controlled release of TGF‐β1, as measured by enzyme‐linked immunosorbent assay (ELISA), was observed with chitosan microspheres over 7 days. Chitosan solutions (2% and 3%) were fabricated into two types of scaffolds with different pore morphologies and mechanical properties using a freeze‐drying technique, with the result that scaffold with higher concentrations showed smaller pores and lower porosity, leading to a much stronger scaffold. The TGF‐β1 microspheres were incorporated into the scaffolds at a concentration of 10 ng TGF‐β1/scaffold and then chondrocytes seeded into each scaffold and incubated in vitro for 2 weeks. The 2% chitosan scaffolds showed higher cell attachment levels than the 3% chitosan scaffolds ( P < 0.01), regardless of the TGF‐β1 microspheres. Both the proliferation rate and glycosaminoglycan (GAG) production were significantly higher for scaffolds incorporating TGF‐β1 microspheres than for the control scaffolds without microspheres 10 days after incubation. Extracellular matrix staining by Safranin O and immunohistochemistry for type II collagen both significantly increased in scaffolds containing TGF‐β1 microspheres. These results suggest that the TGF‐β1 microsphere incorporated in scaffolds have the potential to enhance cartilage formation.