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The injury of articular cartilage is common in clinic and it often caused joint pain, stiffness and dysfunction. Because of cartilage cells embedded in dense matrix, lacking of a direct blood supply, it is self-repair after injury is limited. There are a lot of methods to repair articular cartilage defects in clinical practice, but the result is not satisfactory. Researches[1-4] showed that the low-temperature frozen osteochondral allograft transplantation has much advantages, such as a wide array of sources, long hours of preservation, low immunogenicity, biological activity, binding sites receptor biologically and flexible time limits of transplantation. Therefore, it is subject to a number of researchers’ attention. By observing cartilage morphology, changes of cell metabolism of specimens in each group and the survival status and metabolic activity of the graded cryopreservation on the rabbit articular chondrocytes, to explore the repair effect of articular cartilage.

This study showed that the articular cartilage could tolerate cryopreservation process, and the survival cartilage cells could maintain normal morphology, as well as the synthesis of matrix proteoglycans and collagen type Ⅱ capacity. However, in the view of the histology and ultratructure of frozen cartilage, there were differences among the three groups in different frozen conditions. We find that the changes of cartilage cells in middle layer were the most obvious in morphological and most of cartilage lacuna collapsed, the number of cartilage cells and content of matrix PG and type Ⅱ collagen reduced with passage of time. The differences of which between surface and cartilage matrix increased. This resulted from: ①The cryoprotectant permeability is limited. If not perforated, deep cartilage cells and matrix structure were lack of effective protection and vulnerable to the solution effect injury and mechanical damage. ②The cooling rate of the surface and deep layer of cartilage can lead to inconsistencies in the various parts of the cartilage to form ice crystals. When extra cellular fluid crystall rapidly and cells have not yet solidified, the obvious imbalance of cell osmotic pressure inside and outside can cause dramatic change in cell electrolyte concentration, and lead to cartilage cell death eventually[5-7].
Some researches[8] showed that once given suitable condition for repairing cartilage, the cartilage cells can be differentiated and restore its original features. If the cartilage cells in the middle layer exposed air and increased cryoprotectant penetration to the deep, which can also survived as the surface. The experimental results showed that the experimental group was better than the cryopreservation group. The staining of matrix was higher than the control. It suggested that the effect of cryoprotectants protecting the deep cartilage tissues get significant strengthen, which can improve the survival of cartilage cells effectively.
In addition, we observed an important thing in different periods of cartilage tissue sections stained HE. With the passage of time after transplanted, cartilage cells decreased gradually. However, active cell proliferation situation emerged in the middle layer and superficial partial of cartilage and large cell mass formed. This phenomena in non-hole graded freezing group and cryopreservation group appeared earlier. The phenomenon maybe result from cartilage tissue frozen suffered more serous injury, and the decrease in the number of cells broken the balance of matrix secretion and metabolic. Thereby, it stimulated the proliferation of survival cartilage cell in order to maintain the normal metabolism of cartilage matrix. This is a hypertrophic compensatory mechanism of chondrocytes.
This study showed that the results of cryopreservation group were the worst. The experimental group was better than results of non-hole graded freezing group (P < 0.05). But the number of cartilage cell in the experimental group and non-hole graded freezing group had no significant difference at the same time point (P > 0.05). Which reflected that the disfuntion of the cartilage cell in non-hole graded freezing group was more obvious. Although the changes in cartilage matrix of the experimental group were less than the control, the cartilage matrix and cell inevitably reduced with the passage of time, and the transplanted cartilage degenerated. There are following reasons: ①It was blinded in drilling process, and the appropriate aperture and density of cryoprotectants permeate was unclear. ②The survival cartilage cells were still little after cryopreservation, and it cannot meet the normal metabolism of cartilage matrix. ③Frozen preservation may affect the metabolic activity of cartilage cells. Although cell survived, it was not enough for the synthetisis of matrix. ④The matrix components secreted by transplanted chondrocytes has large differences with the surrounding host cartilage. This maybe affected its biological characteristics[9]. The above reasons maybe directly impact the effect of cartilage transplantation, which should be addressed first and foremost in our future study. In addition, we also observed that the survival of cartilage graft was individual specific. It has significant differences in different individual transplantation in the same conditions. And it had no apparent correlation with the freezing injury. Experimental studies of Steveson[10] showed that the result of histocomparibility leukocyte antigen matching cartilage transplanted was better. However, whether the cartilage transplantation requires histocomparibility antigen matching, we need further study to be confirmed.
In summary, cartilage tissue, as a special type of organization structure, can withstand cryopreservation and has biological activity after transplantation. This provides the necessary preconditions for clinical application. Holed cartilage can protect the deep cartilage cell effectively, improves the survival rate and the effectiveness of transplantation.

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