The Effect Of Cartilage Graft Size On Survivalm Rate Of Chondrocytes And Biomechanical Properties In Tissue Culture

ObjectiveTo provide the experimental evidence for searching the preservation and cartilagetissue medium,choose the articular cartilage in different size of goats to compare thedifference of cartilage cells survival rate and biomechanical properties by preserving themin tissue medium and to oberserve the the influence of TSMU-1medium on cartilage cellsviability.Meterials and MethodsThe articular cartilages of goats were choosed as the experimental material.Underaseptic conditions,obtain cylindrical articular cartilage100,10mm in length and6mm indiameter,of condyleoffemur articular surface by ARTHREX transplantation device,thenwashed by PBS3times.These articular cartilages were equally divided into2groupsrandomly:A1group:the TSMU-1tissue medium (the No.1cartilage preservation ofTaishan Medical College)--added the factor to improve the cell viability in common tissuemedium (the confection method and element are protected by patent);B1group:thecommon medium--99%EMEM tissue medium100UI/ml and1%streptomycin100UI/ml.They were preserved under the condition of4℃,and change the medium everytwo days.Select randomly20cartilages from A and B groups at the first day and35thdayto detect the survival rate of cartilage cell by double fluorescence method-EB/FDA.At the35thday,choose10cartilages from A1group to process into the sample which was6mm inlength,4mm in width,2-3mm in thickness by special bone knife,then,to carry on theunidimensional stretch experiment in automatic control electronic testing machine ofSHIMADU AG-10TA.The rate of stretching is5mm/min.Observe some index such as themaximum load of strech、extension ratio、stress、strain、modulus of elasticity、stress strain curve and so on.Under aseptic conditions,obtain one half of femoral condylar of goats20byhacksaw,they were equally divided into2groups randomly and preserved into A,Bmediums--A2(TSMU-1tissue medium and half of femoral condylar),B2(the commonmedium and half of femoral condylar).In the same condition,to get the whole of femoralcondylar of goats20and also preserved into the two mediums--A3(TSMU-1tissuemedium and the whole of femoral condylar),B3(the common medium and e whole offemoral condylar).Choose20cartilages respectively from the4groups above at the firstday and the35thday.Get10cartilages randomly from A2and A3groups at the35thday toprocess into the sample which was6mm in length,4mm in width,2-3mm in thickness,thento do the same experiment as the A1group to compare the maximum load of strech、extension ratio、stress、strain、modulus of elasticity、stress strain curve and so on.The rateis also5mm/min.All the data were analyzed with the SPSS17.0software and were expressedbyx s.The statistical data of cell livability of all groups were treated by ANOVA.Thenchoose the treatment factor (A:different tissue dmem,35days;B:different size ofcartilages,35days.) that had statistical significance and were tested by One-WayANOVA-q test (The S-N-K way).The data of the unidimensional stretch experiment weretested by One-Way ANOVA-q test.ResultsThe survival rate of chondrocytes of all groups was over90%at the first day,butevery two groups had no statistical significance（P＞0.05）.The cell viability of all groupsreduced at the35thday.In all groups of A group,A1group＞70%，the followed were A2and A3groups,all groups had statistical significance(P＜0.05).About B group,B1groupnearly was50%,both B2and B3groups were lower50%and B2＞B3.The result showsthat the preserving of columnar cartilages is better.The preserving of half of femoralcondylar is better than the whole of femoral condylar.(the data were showed in chart1)By comparing the effect of A and B tissue mediums on the survival rate ofchondrocytes, the result shows:A1group＞B1group，A2group＞B2group，A3group＞B3group.Every two groups had statistical significanc（eP＜0.05）.The result shows that thepreserving of TSMU-1tissue medium is better than the common dmem.(the data wereshowed in chart1) By carring on the unidimensional stretch experiment of A group,the result shows:A1group＞A2group,A1group＞A3group, A2group＞A3group.The result shows thatbiomechanical properties of columnar cartilages is better and the preserving of half offemoral condylar is better than the whole of femoral condylar.(the data were showed inchart2)ConclusionThe different size of cartilage grafts can influence the survival rate of chondrocytesand biomechanical properties in tissue culture in vitro,the effect was better in the smallercartilages. TSMU-1tissue culture medium can increase the survival rate of chondrocyte.MeaningThe repair methods of cartilage defects is still a difficult problem for sportsmedicine and orthopaedics doctors.At present,using the fresh allogenic cartilagetransplanting to treat cartilage defects of full thickness had got certain effect,but thematerials were difficult to get,so it limit the clinical application of allogenic cartilagetransplanting.Therefore,prolonging the preserving time and remaining cells viabilitybecome the focus for scholars.In this study,we found that the presrvation effect was betterin the smaller cartilages.What’s more,the TSMU-1medium developed by SportsMedcinestudy group of Taishan Medical College can increase the survival rate and activity ofcartilage cells and provide the basis for the better preservation and tissue medium.