Cell Therapy To Growth Plate Injury

Objective To develop a simple and reliable method of separation and purification of chondrocytes in vitro.Methods Chondrocytes were obtained from rabbit iliac crest cartilage with aseptic operation,and digested by0.1%hyaluronidase,0.25%trypsin and0.1%type Ⅱ collagen enzyme, then cultured in DMEM/F12supplemented with10%fetal bovine serum.The cells were identified by trypan blue staining.Results Large quantities of individual chondrocytes were obtained with this method with good viability.Conclusions This might be a simple and reliable method for separation and purification of chondrocytes that can be used for further research for cartilage or growth plate injury and repair through amplification by primary culture. Objective To develop a simple and reliable method of primary culture of chondrocytes in vitro.Methods Chondrocytes were obtained from rabbit iliac crest cartilage with aseptic operation,and digested by0.1%hyaluronidase,0.25%trypsin and0.1%type Ⅱ collagen enzyme, then cultured in DMEM/F12supplemented with10%fetal bovine serum.The cells were identified by phase-contrast microscope,toluidine blue staining,type II collagen immunohistochemical staining and transmission electron microscopy.Results Morphologic characteristics of chondrocytes were observed in cell cycle of7-8days.Toluidine blue staining and type Ⅱ collagen immunohistochemical staining showed positive phenotype features of chondrocytes.Transmission electron microscopy showed cytoplasm was rich in mitochondria and endoplasmic reticulum,part of endoplasmic reticulum were expansion.Conclusions This might be a simple and reliable method for primary culture of chondrocytes that can be used for further research for cartilage or growth plate injury and repair on autologous transplantation. Objective To develop a simple and reliable method of growth plate injury model.Methods Growth plates of bilateral proximal tibia from New Zealand rabbit were destructed with needle-pricking with aseptic operation after intraperitoneal anesthesia. The control group was without operation. Injury and repair of growth plates were evaluated by radiological examination.Results1week after the operation, operation group showed local inflammation in bilateral proximal tibial growth plate;6weeks after the operation, operation group showed bone bridge formation in the center of bilateral proximal tibial growth plates.Conclusions This might be a simple and reliable method for establishment of growth plate injury model that can be used for further research for growth plate injury and repair on autologous transplantation with chondrocytes.