ABSTRACT
This study was to assess collagen type II and collagen type I gene expression in tissue-engineered human auricular: cartilage formed via tissue engineering technique. Large-scale culture expansions were transformed into 3D in vitro construct and were implanted subcutaneously on the dorsal of athymic mice. After 8 weeks, explanted construct was processed in the same manner of native cartilage to facilitate cells for gene expression analysis. Isolated cells from in vivo construct demonstrated expression of type II collagen gene comparable to native cartilage. This study verified that tissue-engineered auricular cartilage expressed cartilage specific gene, collagen type II after in vivo maturation.
Subject(s)
Actins/genetics , Cartilage/transplantation , Cellular Senescence/physiology , Cells, Cultured , Chondrocytes/cytology , Collagen Type I/genetics , Collagen Type II/genetics , Ear, External , Fibroblasts/cytology , Gene Expression/physiology , Mice, Nude , Phenotype , Reverse Transcriptase Polymerase Chain Reaction , Tissue Engineering/methodsABSTRACT
Cartilage is regularly needed for reconstructive surgery. Basic research in tissue engineering is necessary to develop its full potential. We presented here the expression profile of type II collagen gene and type I collagen gene in human auricular monolayer culture expansion. Cultured chondrocytes documented a reduction in the expression level of collagen type II gene whilst collagen type I gene was gradually expressed through all the passages. This study demonstrated that human auricular chondrocytes lose its phenotypic expression during monolayer culture expansion. Further studies are required to enhance cartilage specific gene expression, collagen type II throughout the in vitro culture.
Subject(s)
Cells, Cultured , Chondrocytes/cytology , Collagen Type I/genetics , Collagen Type II/genetics , Ear, External , Fibroblasts/cytology , Phenotype , Reverse Transcriptase Polymerase Chain Reaction , Tissue Engineering/methodsABSTRACT
Treatment of articular cartilage lesions remains a clinical challenge. The uses of prosthetic joint replace allograft and/or autograft transplant carry a risk of complications due to infection, loosening of its component, immunological rejection and morbidity at the donor site. There has been an increasing interest in the management of cartilage damages, owing to the introduction of new therapeutic options. Tissue engineering as a method for tissue restoration begins to provide a potential alternative therapy for autologous grafts transplantations. We aimed to evaluate how well a tissue engineered neocartilage implant, consist of human articular chondrocytes cultured with the presence of autologous serum and mixed in a fresh fibrin derived from patient, would perform in subcutaneous implantation in athymic mice.
Subject(s)
Biomechanical Phenomena , Cartilage, Articular/injuries , Cartilage, Articular/physiology , Cartilage, Articular/transplantation , Chondrocytes/cytology , Culture Media , Mice, Nude , Orthopedic Procedures , Serum , Tissue EngineeringABSTRACT
Culture media supplemented with animal serum e.g. fetal bovine serum; FBS is commonly used for human culture expansion. However, for clinical application, FBS is restricted as its carry a risk of viral or prion transmission. Engineering autologous cartilage with autologous human serum supplementation is seen as a better solution to reduce the risk of transmitting infectious diseases and immune rejection during cartilage transplantation. The purpose of this study is to establish and compare the effects of 10% autologous human serum (AHS) and 10% FBS on the growth of chondrocytes and the formation of tissue engineered human articular cartilage.