Experimental Study Of Combined Therapy In Order On The Expression Of BFGF And Tissue Morphology In The Cartilage Of Knee Osteoarthritis Rabbits

Objective:Combined Therapy In Order (joints flushing, injection of hyaluronic acid, iontophoresis of Chinese medicine) is used to treat rabbits'knee osteoarthritis in the experiment. We inspect the expression of bFGF in the cartilage of the right femoral medial condyle and observe tissue morphology in order to clarify the initial mechanism on Combined Therapy In Order which was performed on arthroscopic knee joint. Then to provide theoretical and experimental basis for clinic while using arthroscopic knee on flushing, intra-articular injection of hyaluronic acid and Chinese medicine treatment of iontophoresis.Method:Seventy healthy adult male and female rabbits, about 2.5kg (±100g), which were randomly divided into seven groups:normal controlled group (Ⅰ), model controlled group (Ⅱ), joint flushing group (Ⅲ), hyaluronic acid group (Ⅳ), iontophoresis of Chinese medicine group (Ⅴ), Guyouning embrocation Group (Ⅵ), Combined Therapy In Order group (Ⅶ),10 in each group. A model method of ligation right side of femoral vein is used to get high-pressure. Eight weeks later, the corresponding treatment was used on each group except the normal group and the model group. Five weeks later, the animals were executed. Knees were anatomized, and surface of rabbits'articular cartilage were observed, then a portion of the right femoral condyle cartilage is cutted. According to the sample preparation methods for specimens, the expression of bFGF in cartilage was inspected by immunohistochemical method. And the cartilage cellular ultrastructure was observed with transmission electron microscope.Results:1. BFGF expression in articular cartilage:The treatment groups bFGF expression compared with the model ones have increased significantly (p<0.01), one of the most significant the combined therapy in order group, compared with other treatment group were significantly different (p<0.05).2. Observation the cellular ultrastructure of cartilage by visual observation and transmission electron microscopic: Through visual observation:Ⅰgroup were no markedly abnormal findings in the articular cartilage, articular surface finishing and color as usual.Ⅱgroup:There were obviously changethe in the articular cartilage. The articular cartilage turned yellow obviously and had erosioned, middle layer defected. And the articular cartilage had ulcered, full-thickness defected.Ⅲgroup,Ⅳgroup,Ⅴgroup,Ⅵgroup:The articular cartilage turned roughness,and had a small number of cracks and turned dark color. Combined Therapy In Order group:The surface cartilage than the model group significantly reduce damage. Close to normal group. Through transmission electron microscopic:Ⅰgroup:The chondrocyte had normal eumorphism, complete nucleus, clear nuclear membrane. And chromatin of intranuclear had homogeneous distribution. Nucleolus and organelles in the cytoplasm had normal eumorphism.Ⅱgroup:Apoptosis cells were observed. They showed cell pyknosis obvious, chromatin of intranuclear concentrated and stained darker. Nucleolus reduced or disappeared and organelles in the cytoplasm reduced or even disappeared. There were vacuoles and lipid droplets in cell membrane.Ⅲgroup,Ⅳgroup,Ⅴgroup,Ⅵgroup:The chondrocyte were irregular in shape. chromatin of intranuclear which were basically normal and shallowly stained, and slightly dilated endoplasmic reticulum.Ⅶgroup; The form of chondrocyte presented the circular or ovals were almost normal.The cell membrane had integrity cell morphology. The shape of nucleus became regular. There were plentiful rough endoplasmic reticulum and Golgi complex and mitochondria within the cytoplasm. Through the visual and transmission electron microscopic observation of Combined Terapy In Order group, comparing with other treatment group, it could improve the rabbit knee joint cartilage damage better.Conclusion:Combined Terapy In Order played a very good effect on treating osteoarthritis of the knee. And it is superior to each treatment group. The experimental results show that Combined Terapy In Order has better renovated the rabbit knee cartilage damage compared with the other treatment groups, ameliorated the degeneration of articular cartilage. Combined Therapy In Order provide prtheoretical and experimental basis for clinic while using arthroscopic knee on flushing, intra-articular injection of hyaluronic acid and Chinese medicine treatment of iontophoresis.