Fibrin glue mixed with gelatin/hyaluronic acid/chondroitin-6-sulfate tri-copolymer for articular cartilage tissue engineering: the results of real-time polymerase chain reaction.
Autologous fibrin glue has been demonstrated as a potential scaffold with very good biocompatibility for neocartilage formation. However, fibrin glue has been reported not to provide enough mechanical strength, but with many growth factors to interfere the tissue growth. Gelatin/hyaluronic acid/chondroitin-6-sulfate (GHC6S) tri-copolymer sponge has been prepared as scaffold for cartilage tissue engineering and showed very good results, but problems of cell seeding and cell distribution troubled the researchers. In this study, GHC6S particles would be added into the fibrin glue to provide better mechanical strength, better cell distribution, and easier cell seeding, which would be expected to improve cartilage regeneration in vitro. Porcine cryo-precipitated fibrinogen and thrombin prepared from prothrombin activated by 10% CaCl(2) solution were used in two groups. One is the fibrin glue group in which porcine chondrocytes were mixed with thrombin-fibrinogen solution, which was then converted into fibrin glue. The other is GHC6S-fibrin glue in which GHC6S particles were added into the thrombin-fibrinogen solution with porcine chondrocytes. After culturing for 1-2 weeks, the chondrocytes cultured in GHC6S-fibrin glue showed a round shape with distinct lacuna structure and showed positive in S-100 protein immunohistochemical stain. The related gene expressions of tissue inhibitor of metalloproteinases-1, matrix metalloproteinase-2, MT1-MMP, aggrecan, decorin, type I, II, X collagen, interleukin-1 beta, transforming growth factor-beta 1 (TGF-beta1), and Fas-associating death domain were checked by real-time PCR. The results indicated that the chondrocytes cultured in GHC6S-fibrin glue would effectively promote extracellular matrix (ECM) secretion and inhibit ECM degradation. The evidence could support that GHC6S-fibrin glue would be a promising scaffold for articular cartilage tissue engineering.
Chou
Cheng-Hung
CH
Institute of Biomedical Engineering, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
Cheng
Winston T K
WT
Kuo
Tzong-Fu
TF
Sun
Jui-Sheng
JS
Lin
Feng-Huei
FH
Tsai
Jui-Che
JC
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology*; D018929:Cell Culture Techniques; D019902:Chondrocytes / Q000166:cytology; D002809:Chondroitin Sulfates / Q000737:chemistry; D005109:Extracellular Matrix / Q000378:metabolism; D015718:Fibrin Tissue Adhesive / Q000737:chemistry* / Q000627:therapeutic use; D005780:Gelatin / Q000737:chemistry; D020869:Gene Expression Profiling; D006820:Hyaluronic Acid / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*
D015718:Fibrin Tissue Adhesive; D005780:Gelatin; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates