The Observation on Effect of Traditional Chinese Medicine Gumibao Treating Steroid-induced Necrosis of Femoral Head
|School||Nanjing University of Traditional Chinese Medicine|
|Course||TCM orthopedics and traumatology|
|Keywords||Steroid-induced Avascular Necrosis of the Femoral Head Traditional Chinesemedicine Gumibao Lipid Metabolism Bone Metabolism Biomechanics|
Objective:The purpose of this study is to establish an animal model with clinical pathological characteristics, the disorder of lipid metabolism, intravascular coagulation, bone disorders, osteoporosis in the steroid-induced avascular necrosis of the femoral head in the pathological course of action. And focuses on eliminate the Stagnant and Activate the Blood, Invigorate the Kidney and Replenish Qi-the Effect of Chinese Medical Herb Gumibao on steroid-induced avascular necrosis of the femoral head model of rat femoral head bone tissue morphology, ultrastructure, lipid metabolism, angiogenesis, bone resorption, bone formation effect. To explore the mechanism and efficacy of Chinese medicine, to provide scientific theoretical basis for the treatment of this disease.Methods:Two-month-old SD rats were200of clean grade, half male and half female. Randomly selected30rabbits as blank control group. Molding method according to the S. Okazaki et al, the rat tail vein injection of LPS on two consecutive days of2mg/kg, once a day, the last time after injection of LPS, three consecutive days of injection of methylprednisolone20mg/kg, once a day. The last injection of methylprednisolone after2weeks from the blank control group, model group respectively were6, pathological observation, confirmed that the model was successful. Randomly divided into blank control group (group A), model group (group B), western medicine group (Fosamax Group C), the Gumibao of high-dose group (D), bone mineral density by weight based on Celebrexprescription dose group (E), the Gumibao of low-dose group (F), n=8, male and female. The Gumibao of is high, medium and low dose group rats with body surface area conversion the shares secret Celebrex were orally suspension, western medicine group rats with body surface area conversion gavage dose of Fosamax suspension model groupand blank control group were fed with normal saline.4,8and12weeks after operation respectively intervention.By hematoxylin and eosin staining was used to observe the morphology, observation of morphological changes of femoral head cartilage and bone cell toluidine blue staining, to observe the changes in the number of osteoclasts and TRACP staining, transmission electron microscopy ultrastructural observation, observation of angiogenesis, VEGF staining, Micro-CT scan, biomechanical testing, CHOL, TGL, HDL, LDL detection of lipid metabolism, PICP, CTX-I,1,25-(OH) to detect the level of-D in bone metabolism.Results:The animal model of animal in the injection after LPS died of anaphylactic shock in17, Gumibao group gavage killed during3, western medicine group by gavage killed during2,2rats died in model group and spontaneous.HE staining showed that the trabecular bone of the model group sparse arrangement, shape tapering gradually continuity interruption, bone marrow tissue steatosis, bone karyopyknosis, while poly, stained bone marrow edema, visible fat emboli, a large number of lacunaevacuoles.Blank control group with normal morphology of the femoral head. Western medicine group with normal trabecular bone structure, less breakage, rare lacunae, bone cell shrinkage, nuclear margination. Gumibao group:the high-dose group trabecular bone morphology slender, still arranged neatly, breaking more, a large number of lacunae, bone karyopyknosis, while poly. In the low dose group, trabecular bone structure was normal, nuclear pyknosis, chromatin is less, the bone lacunae number is less, have rich blood vessels under the cartilagie.Toluidine blue staining showed the model group chondrocytes irregular, disorder, the surface cartilage severe disability dye, epiphyseal line at the matrix uneven dyeing, coloring shallow, the osteoblasts irregular, nuclear condensation, nuclear staining light blue, cytoplasmic staining light, epiphyseal plate vacuoles formed around the trabecular bone cells reduce the number. The western medicine group cartilage surface coloring is deep, cartilage cells arranged in neat rows, settling time clearly, nuclear staining evident. Gumibao group uniform staining of the surface cartilage of the femoral head, clear, deep coloring of the surface of the cartilage, cartilage cells arranged in neat rows, clear Shen times, nuclear staining, the presence of bone cells of empty lacunae, the nucleus margination, condensation, trabecular bonearranged in neat rows, the trabecular part of the fracture.VEGF expression located in the cytoplasm and membrane, the positive was brown. Model group after2weeks femoral head VEGF expression was decreased,4weeks,8weeks, although the expression of VEGF within12weeks of the femoral head elevated, but not yet returned to normal levels. Blank control group, VEGF expression did not change. Gumibao group after4weeks of drug intervention based found weak expression of VEGF in the femoral head, the enhanced expression of VEGF in the femoral head within8weeks,12weeks, gradually returned to normal levels.TEM ultrastructure observation:blank control group, bone cells, cartilage cells with normal morphology. The model group appeared in the cytoplasm of lipid droplets, and gradually increase, full of cytoplasm; bone cell nuclear pyknosis, chromatin; bone cell calcification; nuclear and cytoplasmic edema radiolucent zone; trabecular fold, has obvious calcification adhesive line and the newly formed bone; intravascular red cell overlapping even a "Min line", closely arranged, thrombosis.TRACP staining shows, model group TRAP-positive multinucleated cells increased significantly. Around the western medicine group subtidal and trabecular scattered osteoclasts multiple irregular nucleus in a mature or immature shape. Gumibao groups subtidal and trabecular bone scattered around the more mature or immature multicore irregular osteoclast cells. Micro-CT bone morphogenetic3D analysis shows that the model group, BMD, BMC, TMC, TMD, BV/TV to reduce, Tb.SP, SMI, BS/BV increases, share the same Gumibao group, middle dose group compared statistically difference (P<0.05), with Gumibao of the low-dose group and western medicine group difference was significant difference (P<0.01), shares close Celebrex prescription, low-dose group compared with the western medicine group was no significant difference (P>0.05). Micro-CT scan showed normal development of the femoral head of the control group. The rough surface of the femoral head of the model group, the gradual emergence of cellulite degeneration the, early epiphyseal cartilage District cystic change, in the late subchondral bone collapse. The western medicine group and bone trabecula was sparse, the thickness of different sizes, arranged neatly, consistent with the line of force direction. Gumibao groups femoral head surface is rough, stock density remains low dosage group of femoral head bone a little subsidence, epiphyseal plate better continuity, the trabecula was sparse, arranged neatly, consistent with the line of force direction.Serum TC, TG, HDL, LDL shows, model group, TC, TG, HDL was significantly higher HDL reduced, with the other groups was statistically significant (P<0.05). Gumibao groups compared with the western medicine group has no statistically significant difference (P>0.05).High levels of serum PICP content and western medicine group, the Gumibao groups of high-dose and low-dose group were significant difference (P<0.001). The Gumibao groups of high-dose group content of serum PICP with the western medicine group was not statistically significant (P>0.05). The model group CTX-I content is the highest, with the control group, the Gumibao groups of high, middle dose group and western medicine group were statistically significant (P<0.05), the Gumibao groups of low-dose group CTX-I content than western medicine group (P<0.05); model group1,25-(OH)-D content was the lowest, with the blank control group, a statistically significant difference (P<0.05), and western medicine group difference was statistically significant (P<0.001), and there is a significant difference (P<0.01) in the high dose of Gumibao groups.Biomechanical test results showed that the Gumibao groups femoral head subchondral bone maximum load, stress intensity, axial stiffness, elastic modulus significantly better than the model group (P<0.001), Gumibao groups femoral headsubchondral bone displacement values than the model group, two more significant difference (P<0.001). Femoral shaft three-point bending test shares Gumibao groups of various biomechanical indicators are better than the model group (P<0.01).Conclusions:The disorder of lipid metabolism, angiogenesis, reduce the number of bone metabolism imbalance, osteoporosis may be the pathogenesis of steroid induced necrosis of femoral head. Gumibao Fang can regulate lipid metabolism, promote the formation of new blood vessels, accelerate bone repair, inhibit bone resorption, improving biomechanics, delay the collapse of femoral head.