The Experimental Research On The Repair Of Auricular Cartilage Defects By The Complex Of Calcium Alginate And Autologous Chondrocyte

Objective By the transplantation of the complex of Calcium Alginate and Autologous chondrocyte for the repairing of rabbit articular cartilage damage, observing the recently repairing.Methods 1 The articular cartilage cells were digested from rabbit's left knee joint that is 34-37 weeks old and cultured in vitro,and it was easy to get enough quantity of the chondrocytes.2 Counting the number of the chondrocytes, detecting the activity of the chondrocytes by Trypan blue stain, Subculturing and expansion the chondrocytes in vitro. Then identify chondrocyte through immunohistochemical stain of collagen typeâ…¡and Mallory,Toluidine blue stain.3 Low viscosity sodium alginate is dissolved in NaCl solution to made of sodium alginate in the gel, then trickle in CaC12 solution to form the blank alginate beads condensate, for gross observation and HE staining.4 Sodium alginate gel mixed with the third generation of the chondrocytes to form cell gel suspension, and then drops into CaC12 solution to form alginate-chondrocytes complex, and finally culture the cells for 4 weeks in high glucose DMEM medium.5 After cultured for 4 weeks, the calcium alginate-chondrocytes are generally complex morphology, observed under inverted phase contrast microscope, hematoxylin-eosin (HE) staining, and type II collagen immunohistochemical staining.6 Made the model of articular cartilage defects of rabbits, divided it into three groups:the experimental group was implanted the autologous chondrocyte composite, the experimental control group was only implanted the scalfold, the blank group was implanted nothing, the specimen was observed roughly in the eighth week and the twelfth week respectively, HE staining and immunohistochemical stain of collagen typeâ…¡were applied, and then graded score with Wakitani histological grading system.Results 1.(1)The chondrocytes were proliferated rapidly during the three generations and had triangle or multip-angle shape or elliptic shape at the growing confluence, comparing with chondrocyte of the four generations that had irregular morphology, spindle shape, similar to fibroblasts.(2)The positive result was observed with Mallory, staining with toluidine blue and immunohistochemical stain of collagen typeâ…¡.2 The blank calcium alginate and calcium alginate-cartilage cells combined with it by HE staining showed a mesh-like interior, the gap is uniform, with good structure, cells in which the pores were round or spindle within a uniform distribution. Alginate-cartilage cells, by typeâ…¡collagen immunohistochemical staining,shows cytoplasmic and cell typeâ…¡collagen around large.3 Cartilage defects were repaired by hyaline-like cartilage in the experimental group, on the eighth week and the twelfth week, and by fibro-like cartilage repairing in the experimental control group. The blank group were fiber repair at the same time. There was difference between the experimental group and the experimental control group on the eighth week and the twelfth week, there was an obvious statistical difference on with Wakitani histological grading system and They all were better than the blank group.Conclusions 1.The third generation of chondrocytes in cartilage tissue engineering can be used as seed cells for transplant. Chondrocytes within the three-dimensional calcium alginate culture helps to maintain the chondrocyte cell morphology and phenotype.2. Alginate-chondrocytes condensate beads had the function to repair articular cartilage defects, the ability was superior to the scalfold group,and they were better than that of nothing transplanted group.