| Objective: To study the influence of platelet rich plasma(PRP) in reshaping the micro construction and histomorphology of cartilage in joint of KOA, the expression of type ? collagen, Protein polysaccharide, MMP-13 and mRNA by establishing the rabbit model of knee osteoarthritis by Hulth, we try to analyse initially the effect of PRP in treating degenerated cartilage of KOA, probe the potential function mechanis-m of PRP in treating KOA, seek the theoretical basis to applicate PRP clinically. Method: 32 male New Zealand white rabbit, 2.25±0.20 kg weight, were randomly divided into four groups: Group A: blank control group; Group B: model control group; Group C: platelet rich plasma(PRP) treatment group;Group D: sodium hyaluronate treatment group. After one week for adaption, the group A did the simulated operation, while the group B, C, D were mold by Hulth method. After managing the Hulth operation, PRP group,s right knee 3 weeks per time to inject 0.5ml PRP in articular cavity, total 2 times in 6 weeks; sodium hyaluronate group: articular cavity were injected by sodium hyaluronate, once per week and in total 5 times. Group A and B in the same time to give equal amounts of sterile saline injections. 1,Observe the gross appearance of articular cartilage morphological changes and the score, compare the gross appearance shape change between groups. 2,Observe structure of articular cartilage, cell number, the integrity of the tide line and toluidine blue staining, and so on and so forth, compare the Mankin,s score difference between groups. 3,Measure the expression of type ? collagen and GAG of cartilage tissue each group in fluorescent-immunohistochemis- try, compare the differences between groups. 4,Rt-pcr technique was applied to articular cartilage cells of MMP-13 mRNA expression in testing; Using western blot protein imprinting method to detect MMP-13 expression of articular cartilage, compare the differences between groups. Results: 1,compare the general appearance of the articular cartilage: articular cartilage in group A surface is normal; Group B joints, such as the heaviest cartilage injury; C, D group of articular cartilage degeneration but close to group A. According to articular cartilage gross observation standard for evaluation score results: A group the lowest score, B group the highest score. C, D group were significantly lower than those in B group and. A group compared with B group,the difference was statistically significant(P<0.01); Group B and C, D group score comparison, the difference was statistically significant(P<0.01); C group and D group score comparison, the difference was not statistically significant(P>0.05). 2,The articular cartilage were observed under light microscope. Results:in group A, articular cartilage morphology normal; In B group cartilage structure change greatly; C, D Group cartilage morphology has changed, but that with A group cartilage structure is close to that of C, D in the two groups after the treatment of articular cartilage gradually change to the normal structure. The Cell number, tide line and toluidine blue staining of Mankin's score: A group was the lowest.B the highest score of group. C, D group after treatment were significantly lower in. B group and A group score, the difference was statistically significant(P<0.01); B group and C, D group score, the difference was statistically significant(P<0.01), C group and D group were compared, the difference was not statistically significant(P>0.05). 3,The determination results of GAG and type ? collagen: Group B type ? collagen and GAG positive staining of integral optical density(IOD) average is significantly lower in group A; C, D group type ? collagen and GAG positive staining IOD average and A group were closed. A group type ? collagen and GAG compared with B group, the difference was statistically significant(P<0.01); Group B type ? collagen and GAG positive staining IOD average compared with C, D group, the difference was statistically significant(P<0.05); group C type ? collagen and GAG positive staining IOD average compared with the D group, there was no statistically significant difference(P>0.05). 4,The result of MMP-13 protein and mRNA expression in articular cartilage cells: MMP-13 expression of protein and mRNA in the mean value of A group was the lowest, highest in B group, C, D between the two groups. The expression mean of MMP-13 protein and mRNA of A group compared with B group, the difference was statistically significant(P<0.01); Group B MMP-13 and its mRNA expression compared with C, D groups, the difference was statistically significant(P<0.05); MMP-13 expression of mRNA mean comparison of C,D group, there was no statistically significant difference(P>0.05). C, D groups after treatment protein expression of MMP-13 mean comparison, the difference was statistically significant(P<0.01). Conclusion: 1,The rabbit animal model of KOA induced by Hulth method established, possess high success rate and good stability. 2,The preparation of PRP by Landesberg method, the concentration of PRP was relatively stable, and most of the results are the same as those reported in the literature, the method of operation is simple. 3,PRP can promote the repair of cartilage degeneration, reduce cartilage wear, delay the degeneration of articular cartilage lesions in KOA process. 4,PRP can promote the synthesis of type ? collagen and matrix of articular cartilage proteoglycan, promote articular cartilage cell regeneration and repair of cartilaage matrix. 5,PRP can inhibit articular cartilage MMP-13 protein and mRNA expression, effectively prevent the degradation of collagen matrix, delay the disease process, of osteoarthritis protection, inhibit articular cartilage matrix of articular cartilage damage. |
PDF Full Text Request