Study On Blocking SDF-1/CXCR4 Signaling Pathway To Delay The Degeneration Of Human OA Articular Cartilage With TN14003 In Vitro

[Objective] To explore the possibility and mechanism by blocking SDF-1/CXCR4 signaling pathway with TN14003 to prevent the degenetation of OA cartilage, thus provide theoretical basis for the targeted therapy for osteoarthritis(OA).[Methods ]Articular cartilage samples were obtained from patients with OA when acceptted total knee arthroplasty (n = 12) ,the samples were cut into total 140 blocks,which size was 3 ×3×3 mm3, and randomly divided into four groups(group A,B,C,D).Blocks in Group A(n = 12) were incubated in the culture solution contained SDF-1 (100 ng/ml) and TN14003 (1000nmol/L) ,blocks in Group B(n = 12) were incubated in the culture solution contained SDF-1 (100 ng/ml) and T140(1000nmol/L ),blocks in Group C(n = 12) were incubated in the culture solution contained SDF-1 (100 ng/ml) and AMD3100( 1000nmol/L),blocks in Group D(n = 12)were incubated in the culture solution contained SDF-1 (100 ng/ml). The cartilage blocks and culture medium were collected on 2ed,4th,6th,8th and 10th days to take the following tests : ? Histologic examination (HE and Safranin-O stainning);? Q-PCR was used to test the levels of collagen type ?and aggrecan mRNA in the cartilage blocks; ? Western blot was used to test the collagen type II in the cartilage blocks; ?ELISA was used to measure the levels of MMP-3,MMP-9 and MMP-13 in the culture solution. SPSS 19.0 were used with repeated measures analysis of variance for data analysis.The data will be expressed with mean±standard ( X± S) ,P<0.05 was used as siginificant difference standard.[Results].(1) Histologic examination (HE and Safranin-O stainning) of OA blocks:? At the same time,the surface morphology of cartilage blocks in group A showed more intact of the surface,more orderely of collagenous fiber arrangement,more number of chondrocytes,less loss of Safranin-O staining,more orderly peritidal,low Mankin scores than group A, and there was statistically significant (P < 0.05) .? As time going,in the same group the surface morphology of cartilage explants showed graduallydisorder,chondrocyte losing,safranin-O staining losing,mankin scoresgra-dually increasing, and there was statistically significarnt (P < 0.05) .(2) ELISA test the levels of MMP-3,MMP-9 and MMP-13:At the same time , the levels of MMP-3,MMP-9 and MMP-13 was lower in the culture medium of group A than group B,group B was lower than group C, group C was lower than group D,and there was statistically significant ( P < 0.05 ) .As time going,the levels of MMP-3,MMP-13 showed decrease tendency, however, the levels of MMP-9 showed increase tendency.(3)The result of Q-PCR and Western blot in OA blocks:? The result of collagen?and aggrecan mRNA were tested by Q-PCR. At the same time ,the levels of collagen II and aggrecan was lower in the culture medium of group A than group B,group B was lower than group C, group C was lower than group D,and there was statistically significant ( P < 0.05 ) .As time going,the levels of collagen ? Hand aggrecan showed decrease tendency in the same group.?The result of collagen ?protein was tested by Western blot; At the same time ,the levels of collagen ? protein showed decrease tendencyingroupA,B,C,D .and there was statistically significant (P< 0.05 ) . As time going,the levels of collagen II showed decrease tendency in the same group. and there was statistically significant (P < 0.05)[Conclusions] 1.SDF-1 could induce the articular cartilage degeneration in vitro culture.2.TN14003could prevent the articular cartilage degeneration by blocking the SDF-1/CXCR4 signaling pathway.3.TN14003 couldprevent, but could't reverse OA articular cartilage into nomal cartilage.4.TN14003 showed better effect on prevent OA cartilage degeneration by blocking the SDF-1/CXCR4 signaling pathway than T140 and AMD3100.