| Objective:The effects of different formulations of gelatin sponge/fibrin glue scaffolds on chondrocyte migration,proliferation,phenotype,matrix synthesis,and tissue integration after cartilage fragmentation in young rabbits were compared to provide experimental evidence for further research on cartilage fragment transplantation in vivo.Method:Cartilage fragments(approximately 1 mm~3)were obtained from the knee joints of the hind limbs of 36 4-week-old rabbits.The cartilage fragments were implanted into a gelatin sponge scaffold,a gelatin sponge/fibrin glue scaffold,and a fibrin glue scaffold,respectively,and made into a gel-free culture group(group A),a gel-less culture group(group B),and full-gel Culture group(group C)in vitro culture models(36 per group).At 0,2,and 4 weeks of culture,12samples were randomly selected from each group,and the migration,proliferation,phenotype,matrix synthesis,and tissue integration of chondrocytes were observed by histological staining,and the average optical density values of saffron O staining and type II collagen immunohistochemistry were measured.The differences between groups at the same time point and the differences between groups at different observation points were compared and observed.The experimental data were statistically analyzed using SPSS 17.0 software,and the differences between and among groups were compared.Results:1.Histological staining observation:After 2 weeks of culture,a large number of chondrocytes in group A and group B migrated to the space between the fragments and the scaffold material,and more chondrocytes were seen in the fragment in a proliferative state.After 4weeks of culture,chondrocytes in group A and group B were greatly increased.A small amount of new cartilage tissue was formed around the fragments of group A,and the cartilage tissue was partially fused.In group B,there were a large number of chondrocytes and the formation of new cartilage tissue around and between the fragments.The cartilage tissue was obviously fused,and the number of chondrocytes attached to the scaffold material was significantly higher than that of group A.After4 weeks of culture,only a small amount Chondrocytes migrated out,and no fusion was seen in the tissues.2.The average optical density value of safranin O staining:compared between groups,there was no significant difference between the three groups when cultured for 2 weeks(P>0.05),and cultured to 4 weeks,group B was significantly higher than group A and group C(P<0.05)),And there was no significant difference between group A and group C(P>0.05).Comparing within the group,the average optical density value of crocus O staining in each group showed a trend of first decrease and then increase during the period of 4 weeks of culture(P<0.05).When cultured for 4 weeks,group B was significantly higher than 0 weeks(P<0.05),group A was still slightly lower than 0 weeks(P<0.05),and group C was not significantly different from 0 weeks(P>0.05).3.The average optical density value of type II collagen immunohistochemistry:Comparison between groups.At 2 and 4 weeks of culture,group B was higher than group A and group C(P<0.05),while there was no significant difference between group A and group C over the same period(P>0.05).Comparing within the group,the average optical density of type II collagen immunohistochemistry showed a trend of first decrease and then increase during the period of culture to 4 weeks(P<0.05).When cultured for 4 weeks,group B was higher than 0 weeks(P<0.05),and there was no significant difference between group A and group C compared with 0 weeks(P>0.05).Conclusion:The mixed scaffold containing gelatin sponge/fibrin glue can more effectively promote the migration,proliferation,matrix synthesis and tissue fusion of chondrocytes after fragmentation of articular cartilage,and maintain the stable phenotype of exogenous chondrocytes,which is conducive to the repair of hyaline cartilage fragments. |
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