Effects of a Chitosan Scaffold Containing TGF‐β1 Encapsulated Chitosan Microspheres on In Vitro Chondrocyte Culture

Artificial Organs - Tập 28 Số 9 - Trang 829-839 - 2004
Jong Eun Lee1, Seoung Eun Kim2, Ick Chan Kwon2, Hyun Jeong Ahn1, Cho Hyun-Chul1, Sang‐Hoon Lee1, Hee Joong Kim1, Sang Cheol Seong1, Myung Chul Lee1
1Department of Orthopedic Surgery, Seoul National University College of Medicine; and
2Biomedical Research Center, Korea Institute of Science & Technology, Seoul, Korea.

Tóm tắt

Abstract:  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.

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Artificial Organs

Tập 28 Số 9

829-839

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