| Articular cartilage injury caused by trauma or disease is common clinically. But there are not blood vessels and nerves in the cartilage tissues, so self - ple-rosis of the injury is not easy. The plerosis of articular cartilage injury is an important task in the aspects of clinincal and basic research on orthopaedics. How to get large number of autologous chondrocytes is the emphase tesk in the tissue engineering to repair articular cartilage defect. But in previous , the study animals are always the fetal rabbits or immature rabbits'articular cartilage, its'proliferation more fast than the adult rabbits. Many researches show that organism activity gene plays an important role in regulating the mechanism of injury and plerosis of articular cartilage. Many growth factor had been evidenced that their has the effects of promoting proliferation and express the extracellular maxtress. In those, TGF-β1 is a important factor. Clinically,the patients of articular injury or serious osteoarthritis are always the old and adult people, Their self repair capacity is very poor. In present study, we used the adult rabbits articular chondrocytes as the object of study, we researched the effect of TGF-β1 to promote chondrocytes proliferation and preserve the synthetise of special stroma in order to provide theortically and clinically basis of culture and proliferation in vitro of a-dult people articular chondrocytes.Materials and Methods1. Isolation and Culture of adult rabbits chondrocytes: under the sterilecondition, slice the articular cartilage of one Japanese big ear rabbit with the ageof 18 months, the cartilage slices was cut into small cubes (1.0 mm x 1.0 mmx 1.0mm or so) , digested , filtrated, and centrifuged. The collected cells werethen resuspended in DMED culture mediun(containing20% fetal bovine serum, penicillin l00u/ml,streptomycin l00ug/ml) ,adjust the percentage of cells to 2. 5 x 10Vml and cultivated in culture flask at 37 , containing 5% of C02.2. Observe the foim and growth condition of the cells under the invert microscope.3. Observing the cells climbing to the carry sheet glass, which are stained with HE.4. Detect with MTT test to comprehend the survival numbers of the chon-drocytes and the bestest effective concentration of TGF-β1.5. Cell counting and draw growth curve to observe the proliferation state of the chondrocytes under the effect of TGF-β1.6. Detect the expression of n - collagen of the chondrocytes climbing to the carry sheet glass with immunohistochemistry in order to verity the effect of TGF-β1.7. Statistics with analysis of variance.Experimental Results1. Under the invert microscope:chondrocytes began to sticking to the wall of the Petri dish at 4 hours after inoculated and after 48 hours almost all the cells are sticking to the wall. Their phase changed spherical to polygon and some is fusiformis. With the increase of the quantity and volue,cells phase became to be unanimous. The chondrocytes of the test group which be added with TGF-β1 are polygon mostly and have more division cells and blend to be monolayer as "flagstone", the cells of the control group growth slowly and have a few division cells , the proportion of the fusiformis cells increased.2. Observing the cells climbing to the carry sheet glass, which are stained with HE:Under the light microscope, the shapes of chondrocytes may be seen more clearly, being small mostly, in the shapes of fusiform, roundlet and polygon, etc, and karyokinesis may be seen.3 . MIT colorimetric test;Determination of the survival numbers of the cells: 0. D. ( optical density ) value determined by MTT and the survival numbers of the cells are in direct ratio. According to the histogram with various concentrations of TGF-β1, analyzed statistically, the cell nonmember is more super than the control group re-markablely. and the best concentration is l0ng/ml. So we determined that TGF-β1 has the strongest effect of stimulating chondrocyte proliferation and has the be...
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