RESUMO
Objective Chondrocytes were seeded on allogeneic demineralized cancellous bone (DCB ) constructing tissue-engineered cartilage to evaluate the feasibility of DCB as scaffolds for tissue engineering cartilage.Methods Allogeneic DCB was prepared by successive defatting and decalcification and then observed under scanning electronic microscope.One-month old rabbit articular chondrocytes were isolated and proliferated by monolayer culture.Passage 1 chondrocytes were seeded on DCB to construct tissue-engineered cartilage in vitro for 6 weeks.Specimens were taken after 7,14,21,28,35 and 42 days'culture and evaluated by hematoxylin and eosin (HE),toluidine blue (TB),and immunohistochemistry for collagen type Ⅰ and Ⅱ.Results Prepared DCB showed three-dimensional porous structure of 100 - 500 μm with good plasticity and certain mechanical strength. Chondrocytes grew well on and inside the DCB,and were located in lacunae and expressed matrix proteoglycan and type collagen Ⅱ,which formed cartiageous tissues on DCB up to 42 days'culture.Conclusion DCB combined with allogeneic chondrocytes successfully constructed tissue-engineered cartilage in vitro ,which is an ideal scaffold for tissue engineering cartilage.
RESUMO
Objective Chondrocytes were seeded on allogeneic demineralized cancellous bone (DCB ) constructing tissue-engineered cartilage to evaluate the feasibility of DCB as scaffolds for tissue engineering cartilage.Methods Allogeneic DCB was prepared by successive defatting and decalcification and then observed under scanning electronic microscope.One-month old rabbit articular chondrocytes were isolated and proliferated by monolayer culture.Passage 1 chondrocytes were seeded on DCB to construct tissue-engineered cartilage in vitro for 6 weeks.Specimens were taken after 7,14,21,28,35 and 42 days'culture and evaluated by hematoxylin and eosin (HE),toluidine blue (TB),and immunohistochemistry for collagen type Ⅰ and Ⅱ.Results Prepared DCB showed three-dimensional porous structure of 100 - 500 μm with good plasticity and certain mechanical strength. Chondrocytes grew well on and inside the DCB,and were located in lacunae and expressed matrix proteoglycan and type collagen Ⅱ,which formed cartiageous tissues on DCB up to 42 days'culture.Conclusion DCB combined with allogeneic chondrocytes successfully constructed tissue-engineered cartilage in vitro ,which is an ideal scaffold for tissue engineering cartilage.