The Effects Of Shear Stress On The Metabolism Of Human Chondrocyte In Vitro

Objective:Along with our country economy developed at top speed and life-span of population is gradually prolong, the number of patients who suffered osteoarthritis is increased. In our country, the morbidity of osteoarthritis is about 70% in population older than 75. In our country every year the osteoarthritis causes the labor force loses and the expense which produces with the pharmacological treatment to be huge, it also has brought the huge pain for the patient and their family. The number of who . like sports is increased by enhanced of quality of life, but the inappropriate movement is very easy to cause articular cartilage's injury. The articular cartilage injury is quite common in the patient who suffered knee joint disease.The method of therapy injury of cartilage are contain joint arthroscope, drilling, microfracture, autologous chondrocyte implantation, mosaicplasty. For the aim to ACI and let chondrocyte keep its phenotype, so in the first party of my study we have selected the human articular cartilage to carry on the experiment, and we discussion raises and separates the human cartilage cell's method. Although the difficult mechanics function not to have understood in cartilage cell's mechanism, But certain mechanics function has to the articular cartilage cell promotes its secretion cartilage matrix and repairs articular cartilage's function to obtain everybody approval. In the second party of my study the experiment attempts through the different size interrupted cutting devoting to use in the person articular cartilage cell, observed the change of chondrocyte secretion protein polysaccharide and collagen, Interrupted shearing force to cartilage matrix metabolism, the effect of chondrocyte proliferation. It will help us later carry on the recovery exercise,for us the articular cartilage will damage provides the direct basis.Method:The first part of experiment is human articular cartilage which obtains by the surgery in is a material, disappears through two enzymes Person articular cartilage cell's extraction, used high suger DMEM which contain FBS to culture the .chondrocyte, Uses the inverted microscope observation cell growth, Carries on the cell counting with the counting board, than plan cell growth curve. We will gong obtain the human articular cartilage cell undergoes the original generation of raise to multiply next the toluidine blue dyeing examination cartilage cell table. MTT color method estimate cell multiplication situation.The seconds parts of experiments is used the second generation of human chondrocyte which obtains by the first part of experiment. Then let the seconds generation of human chondrocyte arrange to 4 group:control group, low speed group(20rpm), middle speed group(40rpm) and high speed group(60rpm).Simulates the articular cartilage cell to receive the different size the shearing force. The blank control group puts in 5%CO2,37℃incubator to static cultivation. Every day puts the different rotational group human chondrocyte on the rotational table to affect according to the hypothesis rotational speed 2h. (is equal in giving interrupted shearing force), daily two times, previous day replaced the nutrient fluid.Through MTT examination cell multiplication, Rt-PCR examination Aggrecan mRNA and TypeⅡCollagen, the mRNA expression appraises the different size interrupted shearing force to the person cartilage cell multiplication and the secretion protein polysaccharide function.Result:1. Digests the human chondrocyte using two step enzyme digestion method to have the high efficiency at the same time cell harvest to be also big, the original generation of human chondrocyte survival rate which obtains is high.2. The original generation of human chondrocyte undergoes the sugar DMEM culture medium raise to be possible to obtain the cartilage phenotype complete human chondrocyte, it can secrete the protein polysaccharide normally, the cell frozen to the wall is good, may take mechanics function the material.3. The certain intensity interrupted shearing force can obviously affect the human chondrocyte's metabolism, the multiplication activity, causes its synthesis Aggrecan and TypeⅡCollagen increases.4. This experiment's result and clinical and the basic research the understanding which damages regarding the osteoarthritis and the cartilage has the strong relatedness, it is the necessary further deep research. The findings also prompt the treatment which and the recovery stage use nonnegative heavy joint damages in the cartilage exercise (i.e. interrupted shearing force), it possibly obtains a better result.