Effect Of Leptin On Repair Of Articular Cartilage Injury In Rabbits

Objectives The cartilage cells in calcium alginate scaffold co culture and observation of cartilage cells on the scaffolds of biological behavior. The stent implanted in rabbit knee articular cartilage defect model, regular part of knee joint injection 10ng/ml leptin,observation, defect repair, and then discusses leptin on rabbit articular cartilage injury of the knee joint is the repair effect.Methods 1 Isolation and culture of chondrocytes: 2 healthy 4 week old New Zealand rabbit articular cartilage, the chondrocytes were isolated and amplified and passaged. The chondrocyte and star shaped, light polygon, cobblestone growth; observation of cartilage cell morphology HE staining, immunohistochemical staining of type II collagen secretion and sodium alginate scaffolds. 2 The mixture of calcium chloride, the chondrocytes were cultured in the passage. The proliferation state of the cells were observedunder microscope, and HE staining and immunohistochemical staining. 3 The rabbit model of knee joint defect in adult rabbits. 4 The experimental animal were randomly divided into 3 groups: control group: every5 of the rabbit the knee joint cavity injection of 1ml sodium chloride physiological; stent group: the three-dimensional scaffolds containing chondrocytes good rabbit knee joint defect cavity implantation dressing, and every 5 day injection of sodium chloride physiological 1ml;experimental group: three-dimensional stent implantation After every 5 days, and local injection of 1ml of 10 ng / ml leptin. The attention were randomly divided into two groups, the experimen-tal individual defect sites of the same, the specimens were were killed 12 weeks afte-r operation, observe the cartilage defect repair of growth, and stained with hematoxyli-n and eosin, Masson staining, immunohistochemical staining and quantitative PCR analysis of inspection and repair. To evaluate leptin on 3D scaffold of cartilage defect repair.Results 1 Separated cartilage cells were round or oval, after 12 hours after gradually attached to the wall after 24 hours, most have been posted HE cartilage cell wall. 2 After passage staining under microscope were triangular, polygonal or spindle shaped nuclei, obviously,were round or oval, HE staining and immunohistochemical staining results. The chondrocytes showed cartilage cells, its unique activity. 3 Suspension of sodium alginate, calcium chloride was prepared by mixing, scaffold, apoptosis and lysis of stent cultured cells was not obvious,and the cell activity was good, inspection results show that cells suitable for implantation of. 4Articular cavity after implantation, no obvious inflammatory reaction and immune rejection the reaction, support suitable for soft and hard, will not damage the articular cartilage, and the later material degradation completely, no joint residual scaffold. 5 Implantation after dressing and daily injections of leptin, promote collagen type II and IGF-1 students Cheng, the repair of the defect has a significant role in promoting. 6 Experimental group: the knee joint defect surface is smooth, compared with the other two groups, PCR amplification assay of IGF-1secretion was significantly higher than the other two groups, the difference was statistically significant; the control group: the defect was repaired, knee joint adhesion, articular cartilage damage is serious, obviously the performance of osteoarthritis; stent group: the defect of cartilage is smooth, but lower than the experimental group, pathological examination showed no. 7 Knee osteoarthritis, experimental group fibers and less scar tissue, type II collagen staining showed that the number is higher than the other two groups, HE staining showed a few fibroblasts, nuclear small, arranged the relative rules. The control group of fibrous tissue more disordered, there are a few cartilage cells, large nuclei and irregular arranged. The stent group cartilage cell nucleus, between the number of fibroblasts and structure between the two.Conclusions 1 Calcium alginate scaffold and cartilage cells were co cultured, cartilage cell can be very good in the scaffold surface and in the interior of the adhesion, growth and proliferation, cell activity is good, and there is no obvious cytotoxicity. 2 Experimental process in calcium alginate scaffold material in rabbits caused by inflammation reaction was mild, well vascula rized, slow absorption of the material degradation, with good organization compatibility. 3 The experimental conditions under 10ng/ml leptin can promote the proliferation of cartilage cells and type II collagen and IGF-1 secretion,locally injected regularly can promote the repair of rabbit articular cartilage injury of the knee joint.