RESUMO
BACKGROUND:Such methods as transplanting autologous periosteum or autologous bone marrow mesenchymal stem cells (BMSCs) can promote the repair of articular cartilage defects for sure. But they all have their own limits in chondrogenic abilities,which results in an unsatisfactory curative effect.OBJECTIVE:To study the effect of transplanting BMSCs (which were induced into chondrocytes) combined with autogenous periosteum on repairing articular cartilage defects in rabbits.MATERIALS:A total of 18 New Zealand rabbits,aged 6-8 months,were divided with random digits table method into 3 groups,namely,periosteum+BMSCs group,periosteum group and blank control group,with 6 ones (12 knee joint samples) in each group. METHODS:In periosteum+BMSCs group,BMSCs were harvested and adherently cultured with trypsin digestion method. Then they were induced by transforming growth factor 81 into chondrocytes. At the same time,immunofluorescence labeling was performed to BMSCs membranes with PKH-26. Full-thickness articular cartilage defects (diameter:3mm,depth:3mm) were made to bilateral condylus medialis femoris of all rabbits. In periosteum+BMSCs group and periosteum group,defects were covered by homolateral autogenous proximal tibia periosteums,with germinal layer facing to cavitas medullaris. After that,the periosteum+BMSCs group received 3 sutures,followed by injection of 20 μL BMSCs suspension (1×109/L) into the defects,after which the last suture was taken. The periosteum group underwent coverage with periosteum on defects only. The blank control group underwent perforate only.MAIN OUTCOME MEASURES:General observation,histological observation,Wakitani's score,immunohistochemical and in situ hybridization detection of collagen type Ⅱ were performed to defects at 6 and 12 weeks following operation.RESULTS:No sutured periosteums were found desquamate. In periosteum+BMSCs group,defects were filled with hyaline cartilage-like repairing tissues at week 6 following operation;Week 12 following operation saw remodeled tissues whose cells were mainly the implanted cells labeled with PKH-26. In periosteum group,repairing tissues in defect areas were ivory white,smooth with light introcession and distinctively different from the surrounding normal cartilage tissue. In the blank control group,clearer introcession or irregular appearance,even broken surrounding cartilage tissues could be seen in the defect area. Both Wakitani's score and histological score were highest in periosteum+BMSCs group at week 6 and 12 following operation (P＜0.05),with higher ones in periosteum group than in the control group (P＜0.05). What'more,matrix around cells in the repairing tissues showed positive results to both immunohistochemical and in situ hybridization staining of collagen typeⅡ,which proved that cells in repairing tissues were the implanted ones.CONCLUSION:Transplanted BMSCs (which were induced into chondrocytes) combined with autogenous periosteum can form hyaline cartilage-like repairing tissues through which articular cartilage defects are repaired.