| Objective:To observe the expression of MT1-MMP(Membrane type-1 matrix metalloproteinase,MT1-MMP)in minced rabbit articular cartilage in vitro culture,and reserching the correlation of the expression with chondrocyte migration and minced cartilage repaire.analyze their difference,provide experimental evidence for studying the mechanism of articular cartilage healing.Method: full-thickness articular cartilage were obtained form femoral condyle and tibial plateau of 60 knees of 1 months old rabbits.Part I: The articular cartilage was minced into particulate(1×1×1mm),plant it into Fibrin glue-Gelatin sponge composite scaffolds,30 models of cartilage fragements cultured in vitro were finished(group A).Obtaining 30 monoblock cartilage from the condyle of femur(about 7mm in diameter,3mm in thickness)plant into scaffolds as the control group(group B).10 specimens were taken out respectively from each group after cultured 0,2,4 weeks.Chondrocytes at the margin of tissue were observed via HE staining and histologically evaluated with semi-quantitative scoring,the expression of MT1-MMP and Collagen ? were evaluated by Immunohistochmical,and calculate positive cell percentage of MT1-MMP in Immunohistochemistry sections and obtain average optical density values of Immunohistochemistry sections of collagen ?.Compared the difference of the two groups at same time point,and different time point in the same group.Part ?: Manufacturing 30 models of Minceded articular cartilage-Fibrin glue-Gelatin sponge compound else(group C),and cultured in vitro with inhibitor of MT1-MMP(NSC402050),100umol/L;10 specimens were taken out after cultured 0,2,4 weeks.chondrocytes at the margin of tissue were observed via HE staining and histologically evaluated with semi-quantitative scoring.Contrast difference between group A and group C.Analyze: All measurement data were analyzed with Graphpad Prism 6.0,2 Way ANOVA were used in groups to analyze the data of histological semi-quantitative scoring,T Test were used in groups to analyze the data of positive cell percentage of MT1-MMP and average optical density values of Immunohistochemistry sections of collagen ?,Spearman correlation analysis were used to analyze the correlation between positive cell percentage of MT1-MMP and histological semi-quantitative scoring,Pearson correlation analysis were used to analyze the correlation between positive cell percentage of MT1-MMP and average optical density values of Immunohistochemistry sections of collagen ?.Results: Part I: 1.Positive cell percentage of MT1-MMP: 0 week,the average positive cell percentage of MT1-MMP is 17.1%±2.9%.The percentages were increased with time in both groups,multiple comparisons at all time points were made in both group,a significant different was detected at 0st week,2nd week and 4rd week,P<0.05.at 2nd,4th week of culture,the percentage of positive cell of group A is higher than that of group B,a significant different was detected,P<0.05;2.Histological semi-quantitative scoring of HE stain: The scores were increased with time in both groups,multiple comparisons at all time points were made in both group.At 2nd,4th week of culture,the scoring of group A is higher than that of group B(P<0.05);3.Average optical density values of Immunohistochemistry sections of collagen ?: The Average optical density values were increased with time in both groups,multiple comparisons at all time points were made in both group(P<0.05);the average values of group A is higher than that of group B,P<0.05;4.correlation between positive cell percentage of MT1-MMP and histological semi-quantitative scoring: there was a positive correlation between positive cell percentage of MT1-MMP and histological semi-quantitative scoring,a significant different was detected,r=0.774(P < 0.05).5.correlation between positive cell percentage of MT1-MMP and average optical density values of Immunohistochemistry sections of collagen ?: there was a positive correlation between positive cell percentage of MT1-MMP and average optical density values of Immunohistochemistry sections of collagen ?,a significant different was detected,r=0.786,P<0.05.Part ?: 1.Positive cell percentage of MT1-MMP: In the group C,the percentage of MT1-MMP positive cell is very high in 2nd,4th week,about 100%,insignificant,not included in the analysis.2.Histological semi-quantitative scoring of HE stain: In group C,a insignificant different was detected from the comparisons of 0th week vs 2nd week and 2nd vs 4rd week,P>0.05,no chondrocytes gathered at the margin of tissue;compared the group A and group C,at 2nd,4th week of culture,the scoring of group A is higher than that of group C(P<0.05).3.Average optical density values of Immunohistochemistry sections of collagen ?: In group C,the value of 2nd week higher than 0th week(P< 0.05),and a insignificant different was detected from the comparisons of 2nd week and 4th week,P>0.05;the average values of group A is higher than that of group C at the same time point,P<0.05;Conclusion: After the fragmentation of articular cartilage in juvenile rabbits,the expression of MT1-MMP was up-regulated and the migration of chondrocytes increased. |
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