TGF-beta1 immobilized tri-co-polymer for articular cartilage tissue engineering.
Tri-co-polymer with composition of gelatin, hyaluronic acid and chondroitin-6-sulfate has been used to mimic the cartilage extracellular matrix as scaffold for cartilage tissue engineering. In this study, we try to immobilize TGF-beta1 onto the surface of the tri-co-polymer sponge to suppress the undesired differentiation during the cartilage growth in vitro. The scaffold was synthesized with a pore size in a range of 300-500 microm. TGF-beta1 was immobilized on the surface of the tri-co-polymer scaffold with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) as a crosslinking agent. Tri-co-polymer scaffolds with and without TGF-beta1 were seeded with porcine chondrocytes and cultured in a spinner flask for 2, 4, and 6 weeks. The chondrocytes were characterized by the methods of immunohistochemical staining with anti-type II collagen and anti-S-100 protein monoclonal antibody, and RT-PCR. After culturing for 4 weeks, chondrocytes showed positive in S-100 protein, Alcian blue, and type II collagen for the scaffold with TGF-beta1 immobilization. There is no observed type I and type X collagen expression in the scaffolds from the observation of RT-PCR. In addition, the scaffold without TGF-beta1 immobilization, type X collagen, can be detected after cultured for 2 weeks. Type I collagen was progressively expressed after 4 weeks. These results can conclude that TGF-beta1 immobilized scaffold can suppress chondrocytes toward prehypertrophic chondrocytes and osteolineage cells. The tri-co-polymer sponge with TGF-beta1 immobilization should have a great potential in cartilage tissue engineering in the future.
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei 100, Taiwan, Republic of China.
Cheng-Hung
CH
Chou
Winston T K
WT
Cheng
Chien-Cheng
CC
Lin
Chih-Hung
CH
Chang
Chien-Chen
CC
Tsai
Feng-Huei
FH
Lin
D000818:Animals; D040761:Biomimetic Materials / Q000138:chemical synthesis; D002358:Cartilage, Articular; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology; D002809:Chondroitin Sulfates; D005780:Gelatin; D006820:Hyaluronic Acid; D011108:Polymers / Q000737:chemistry* / Q000627:therapeutic use; D016062:Porosity; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*; D016212:Transforming Growth Factor beta / Q000008:administration & dosage*; D053773:Transforming Growth Factor beta1
D011108:Polymers; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D005780:Gelatin; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates