| ObjectiveTo observe the variation of BMP-2 and VEGF as well as patho-change in the steroid induced avascular necrosis of femoral head in rabbits under the effect of Gutong ointment,discussing the control efficiency and mechanism of Gutong ointment on SANFH,to provide experimental basis for the prevention and treatment of SANFH with traditional Chinese medicine.Methods30 male New Zealand rabbits were used to establish the model of steroid induced avascular necrosis of the femoral head by using the improved Lipopolysaccharide combined with methylprednisolone.After the model was completed,using X-ray and MRI identify the model.After the success of the model identification,the experimental animals were randomly divided into three groups:blank group,control group and experimental group.Removing the rabbit hair on both hips,smearing the gutong ointment on the cloth cotton(size about 10*10 cm)and applying on both hips of the experimental group.The experimental group changed dressing every other day and continued 4 weeks.Other two groups were raised normally.After the treatment,all rabbits were put to death.The femoral head was taken for HE staining of tissue morphology and empty bone lacuna rate detection.The expression of BMP-2 and VEGF in femoral head was detected by immunohistochemistry.Results1.All rabbits are in good mental state,with bright and shiny hair,sensitive reaction,normal appetite and defecation before the experiment.After modeling,most of the rabbits were mentally depressed,with a slightly dull and fluffy coat,a little hair removal,less activity,slow reaction,a slight loss of appetite,some of them had diarrhea,and slightly lost weight.MRIT1 showed flaky low signal in the femoral head and T2 lipid inhibition showed flaked heterogeneous signals in the femoral head and hydrops in the hip joint,which confirmed the success of the model.2.The generally morphological observation of femoral head.①Blan k group:The femoral head has complete shape without collapse of cartilage surface,the whole is white with smooth surface,the bone is hard and difficult to cut;②Control group:The femoral head has complete shape,and there is no obvious collapse of the cartilage surface.The surface luster of the femoral head is slightly dim,the whole femoral head is cyan,and the cortex is obviously thinner.The bone is osteoporosis and easy to cut;③Experimental group:The femoral head has complete shape,and there is no obvious collapse of the cartilage surface.The surface of the femoral head is glossy,and the whole bone is pale white with transparent cartilage.The bone is hard and needs some force to cut.3.HE dyeing observation of femoral head.①Blank group:Bone trabeculae arranged in a continuous and orderly manner.Osteocytes are clearly visible and nucleus is large a nd located in the center.There is less fat in the medullary cavity and abundant hematopoietic cells,the empty bone lacuna rate is 12.128%;②Control group:The cartilage of the femoral head becomes thinner.Bone tissue continuous middle,trabecular appear rupture.Osteocytes cell nucleus appeared small and karyopycnosis.Bone cells scattered irregular,the number of osteocytes was significantly reduced and empty lacunae increased.Hematopoietic cells in the medullary cavity was decreased.There were a lot of fat cells in the lacuna,and some fused into vesicles,the empty bone lacuna rate is 12.12819.592%;③Experimental group:Bone tissue continuous fair,trabecular bone slightly thinner,failure is not obvious,visible new bone formation and bone cell distribution rules,intramedullary femoral head visible hematopoietic cells and empty osteocyte nest no significant increased,the empty bone lacuna rate is 15.213%.The results of empty bone lacuna rate were analysized by variance.Compared with the blank group,both control group and experimental group have higher rates and the difference is statistically significant(p<0.05),which proving the success model.There also has significant difference between control group and experimental group(p<0.05).4.Immunohistochemical results:①The expression of BMP-2 and VEGF in control group and experimental group were lower than that in blank group,with statistical significance(p<0.05);②The expression of BMP-2 and VEGF in experimental group was higher than that in control group,with statistical significance(p<0.05).Conclusions1.The method using lipopolysaccharide combined methylprednisolone to establish animal model of steroid induced avascular necrosis of femoral head is simple and reliable,which has low mortality and high success rate of modeling.2.The occurrence and progression of steroid induced avascular necrosis of femoral head are closely related to the expression of BMP-2 and VEGF in bone tissue.BMP-2 and VEGF can promote angiogenesis and new bone formation.3.Gutong ointment can increase the expression of BMP-2 and VEGF in bone tissue of steroid induced avascular necrosis of femoral head and promote angiogenesis and bone repair in the necrotic area,which has some prevention and cure function. |
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