Use of synovium-derived stromal cells and chitosan/collagen type I scaffolds for cartilage tissue engineering.
The objective was to investigate synovium-derived stromal cells (SDSCs) coupled with chitosan/collagen type I (CS/COL-I) scaffolds for cartilage engineering. CS/COL-I scaffolds were fabricated through freeze-drying and cross-linked by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. SDSCs were isolated from synovium and cultured onto CS/COL-I scaffolds, constructs of which were incubated in serum-free chondrogenic medium with sequential application of TGF-β1 and bFGF for up to 21 days and then implanted into nude mice. The physical characteristics of the scaffolds were examined. The quality of the in vitro constructs was assessed in terms of DNA content by PicoGreen assay and cartilaginous matrix by histological examination. The implants of the constructs were evaluated by histological and immunohistochemical examinations and reverse transcription PCR. Results indicated that the CS/COL-I scaffold showed porous structures, and the DNA content of SDSCs in CS/COL-I scaffolds increased at 1 week culture time. Both of the constructs in vitro and the implants were examined with positive stained GAGs histologically and the implants with positive collagen type II immunohistochemically. RT-PCR of the implants indicated that aggrecan and collagen type II expressed. It suggested that SDSCs coupled with CS/COL-I scaffolds treated sequentially with TGF-β1 and bFGF in vitro were highly competent for engineered cartilage formation in vitro and in vivo.
Gong
Zhongcheng
Z
Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830054, People's Republic of China.
Xiong
Hui
H
Long
Xing
X
Wei
Lili
L
Li
Jian
J
Wu
Yang
Y
Lin
Zhaoquan
Z
D000818:Animals; D002356:Cartilage / Q000166:cytology*; D002478:Cells, Cultured; D048271:Chitosan / Q000032:analysis / Q000737:chemistry*; D020219:Chondrogenesis; D024042:Collagen Type I / Q000032:analysis / Q000737:chemistry; D024043:Collagen Type II / Q000032:analysis / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000032:analysis / Q000737:chemistry; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000032:analysis / Q000737:chemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D016062:Porosity; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology*; D013583:Synovial Membrane / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D053773:Transforming Growth Factor beta1 / Q000032:analysis / Q000378:metabolism
D024042:Collagen Type I; D024043:Collagen Type II; D003432:Cross-Linking Reagents; D053773:Transforming Growth Factor beta1; D048271:Chitosan; D005022:Ethyldimethylaminopropyl Carbodiimide