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Objective calcium alginate composite autologous chondrocyte transplantation for the repair of rabbit knee articular cartilage defects, observed defect repair. 1 take 34-37 weeks in New Zealand white rabbits left knee cartilage, digested with high purity chondrocytes in monolayer culture. 2 cultured in vitro cartilage cell line station hope blue-activity detection, amplification passage, and its morphological observation, cell growth curve. In the third generation of cartilage cells seeded row Mallory staining, toluidine blue staining, and Ⅱ collagen immunohistochemical staining to identify chondrocytes; 3. Analytically pure low viscosity sodium alginate (sigama) was dissolved in a solution of NaCl prepared sodium alginate gel and cross-linked to form a blank calcium alginate beads dropped into the CaCl2 solution, gross observation and HE staining. Sodium alginate gel and digested chondrocytes mixed into cell gel suspension was dropped into the CaCl2 solution in the cross-linked to form calcium alginate - chondrocytes Ning Chu, placed high glucose DMEM complete medium Bank of China three-dimensional culture for 4 weeks. The culture after 4 weeks of calcium alginate - cartilage cells Ning Chu by gross morphology of inverted phase contrast microscope, hematoxylin - eosin (HE) staining, and type II collagen immunohistochemical staining. Production rabbit articular cartilage defect model, divided into three groups, the experimental group was implanted autologous chondrocyte - carrier complex, experimental control group soaked in calcium alginate beads implanted in a single medium, the control group implantation materials, a single defect model. Groups, respectively, after 8 weeks, 12 weeks after operation gross observation, HE staining, and type II collagen immunohistochemical staining and histological scores with Wakitani method. 1 (1) the rapid growth and proliferation of the previous three generations of chondrocytes, the shape of a polygon, short spindle, triangle, fusion cobblestone shaped oval, opposite four generations after chondrocytes irregular, fusiform The sixth generation of cartilage cells like fibroblasts. (2) The third generation of cartilage cells Mallory, toluidine blue metachromatic reaction and type II collagen immunohistochemistry strongly positive. 2 (1) blank calcium alginate and calcium alginate - chondrocytes condensate beads HE stain was visible inside the condensate beads mesh-like gap is relatively uniform, with good spatial structure, the cells were round or Ning Chu pore spindle uniform distribution, good growth; (2) calcium alginate - chondrocytes complex type II collagen immunohistochemical staining around the cell cytoplasm and cell type II collagen expression. Animal models after 8 and 12 weeks, the experimental group defects as hyaline cartilage-like repair, repair of experimental control group fibrocartilage-like, fibrous repair of the control group. Repair at the same point in time, the experimental group is better than the experimental control group. Which Wakitani histological score results showed that the experimental group and the experimental control group were significantly different, and both were better than the control group, at the same time in each group at week 12 score results are better than eight weeks. Conclusion The third generation of autologous cartilage cells can be used as seed cells for cartilage tissue engineering transplantation, the three-dimensional culture of chondrocytes in calcium alginate helps to maintain the morphology and phenotype of chondrocytes. Calcium alginate - autologous chondrocyte complex articular cartilage defect repair, repair is better than the stent control group, the difference was statistically significant, and the two sets of results are better than the control group.
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