Effects of the controlled-released TGF-beta 1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold.
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.
Lee
Jong Eun
JE
Seoul National University College of Medicine, Orthopedic Surgery, 28 Yongon-dong Chongno-gu, Seoul 110-744, South Korea.
Kim
Ko Eun
KE
Kwon
Ick Chan
IC
Ahn
Hyun Jeong
HJ
Lee
Sang-Hoon
SH
Cho
Hyunchul
H
Kim
Hee Joong
HJ
Seong
Sang Chul
SC
Lee
Myung Chul
MC
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D020099:Coated Materials, Biocompatible / Q000008:administration & dosage* / Q000737:chemistry; D003094:Collagen / Q000737:chemistry; D003692:Delayed-Action Preparations / Q000008:administration & dosage / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods*; D005109:Extracellular Matrix / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry; D008422:Materials Testing; D008863:Microspheres; D016062:Porosity; D011817:Rabbits; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*; D016212:Transforming Growth Factor beta / Q000008:administration & dosage* / Q000737:chemistry
D020099:Coated Materials, Biocompatible; D003692:Delayed-Action Preparations; D006025:Glycosaminoglycans; D016212:Transforming Growth Factor beta; D003094:Collagen; D048271:Chitosan