| Objective:The comparative analysis of external cartilage differentiation between rabbit bone marrow mesenchymal stem cells(MSC)and Buffy coat which includes mononuclear cells,growth factors and cytokines etc.The aim of this study was to provide experimental basis for the treatment of cartilage injury with Buffy coat.Methods:The selected animals were 16 weeks old,the weight was about(3±0.5)kg,and there were 12 New Zealand white rabbits.All 12 New Zealand white rabbits were randomly divided into three groups:MSC group(negative control group):3 rabbits,apply the cultivation under the induction of non-additive transforming growth factorβ in bone marrow mesenchymal stem cells(MSC).Buffy coat group(experimental group):6 rabbits,apply the cultivation under the cartilage induced liquid without experiencing external continuous cell culture of Buffy coat.MSC+TGF group(positive control group)3 rabbits,apply the cultivation under the induction of adding transforming growth factor(3 in bone marrow mesenchymal stem cells(MSC).Through the gross observation,histological observation,glycosaminoglycan(GAG)assay,and in vitro cartilage BERN score compared the external cartilage differentiation ability.Results:1)The general observation showed:the cartilage part cultivated in MSC group had been dissolved,fuzzy edge;MSC+TGF group and Buffy coat group had no dissolution phe no me non,smooth and clear edge.2)The sarranine O staining microscope morphological observation showed:the chondrocytes in MSC group had poor growth,maldistribution,precious few amount and GAG secretion,irregular secretion;a few cells can find specific Cartilage lacuna of chondrocyte.Buffy coat group and MSC+TGF group had rather fast growth speed of chondrocyte,even cells distribution,cell number was rather large,smooth surface,round and mellow periphery,there are a large number of GAG secretion,basically distributed in the surroundings of chondrocytes;there are a large number of cells found in Cartilage lacuna.3)Glycosaminoglycan(GAG)content determination:MSC group:7.78±1.79μg,Buffy coat group:22.83±3.11μg,MSC group TGF:24.05±3.35μg.There is significant difference in between the three groups,MSC group and MSC group TGF,MSC and Buffy in coat group,there is significant difference between groups(P<0.05);Buffy coat group and MSC TGF group,no significant difference between groups(P>0.05).4)In vitro cartilage BERN score:MSC ggroup:2.83 ± 1.16,Buffy coat group:6.16±1.16,MSC+ TGF group:6.68±0.81.There is significant difference in between the three groups,MSC group and MSC group TGF,MSC and Buffy coat group,there is significant difference between groups(P<0.05),Buffy coat group and MSC TGF group.There were no significant differences(P>0.05).Conclusions:Buffy coat has a good in vitro chondrogenic differentiation ability,which can solve the problem that the growth factor must be added in vitro differentiation cartilage,which provides the experimental basis for the clinical application of Buffy coat. |
