| Osteoarthritis is a disease that the characteristic of which is cartilage degeneration,osteophyte formation and subchondral bone remodeling.The incidence rate of OA steps by each year and had caused great disablity to the old people.A suitable animal model is the basis of OA research.Athouth we have modeled OA by different means,little is known about the detail of these models.The serum component of OA patients is different from normal person and maybe it is the reflection of the painful joint.Inversely,the changed serum environment may have a influence on the progression of OA.In this experiment,we research systematicaly the modified Hulth modeling of OA rabbit,and observe the influence of OA rabbit serum at different stages on the proliferation of chondrocytes and mesenchymal stem cells.Objective:1.To observe the macroscopic and pathologic characteristics of the rabbit knees and survey the NO concentration of the serum,chondrocytes apoptosis, collagenⅡmRNA and aggrecan mRNA change in different phase after Hulth's modeling and confirm the feasibility of replicating knee osteoarthritis at different stages2.To study the influence of OA rabbit serum afer different times postoperation on the proliferation of chondrocytes and mesenchymal stem cells.Method:1.sixteen mature New Zealand white rabbits were subjects of this study.The right knees were selected as the experimental group in which the ACLT of those knees were cut off and medial meniscus resected,and left knees as a normal control group.The rabbits were sent to a large cage,where they were free to move.Four rabbits were killed at the 2nd,6th,10th and 14th weeks,respectively.The macroscopic and pathologic characteristics of the rabbit knees were observed.A histologic evaluation was done according to the Mankin grading scale.We also quantified the NO concentration of the serum and chondrocytes apoptosis between different group and tested the expression level of collagenⅡmRNA and aggrecan mRNA with PT-PCR between different group;2.Isolate and culure OA chondrocytes and compare it with the normal chondrocytes,compare their ability of proliferation by the means of MTT assay3.culture the normal chondrocytes and MSCs with the osteoarthritic serum,compare cells proliferation with different conditioned-culture solution by means of MTT assay.Result:1.The Mankin's scale between the control group didn't have statistical signification(P＞0.05).but the scale between the control and modeling group and beween different modeling group had apparent statistical signification(P＜0.01).The NO concentration of the serum between the normal,2th and 6th weeks modeling group had statistical signification(P＜0.01),but between the 6th,10th,14th weeks modeling group hadn't(P＞0.05).The chondrocytes apoptosis percentage and mRNA expression of collagen and aggrecan between different group had statistical signification(P＜0.01).2.The osteoarthritic chondrocyte have a better ability of proliferation then the nomal chondrocytes at 10th,14th weeks(P＜0.01)3.Osteoarthritic serum conditioned -culture solution can change the shape of the chondrocytes and MSC and its compact on cells proliferation is complicated.Conclusion:(1) The osteoarthritis of the knee at different stages could be successfully replicated by the Modified Hulth' s Modeling Method within 14 weeks,early stage changes occur at 2 weeks,metaphase at the 6th week,advanced stage at the 10th week and last stage at the 14th week.The NO concentration increased rapidly before 6th weeks modeling,and then get into plateau phase.The chondrocytes apoptosis pencentage increased with the histological grading scale;the highest mRNA level of collagen and aggrecan appears at the 10th week and then decline(2) The osteoarthritic chondrocytes seems to have better proliferation potential,we advise we can choose it as the seeding cells of tissure engineering;(3) the osteoarthritic serum has great influence on the biological behavior of chondrocytes and MSCs,and we need deep research.
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