| ObjectiveThe contrast-enhanced ultrasound(CEUS)technique was used to observe the Steroid induced osteonecrosis of the femoral head(SIONFH),by the sonographic characteristics,contrast medium perfusion pattern,and quantitatively analyze the parameters of CEUS,and so on.The main objectives include: ?based on the animal models,the rabbit model of SIONFH was established,the modeling effect was verified from the aspects of histopathology,micro vessel density and hematological indexes in different periods,and CEUS examination was carried out simultaneously to explore the key mechanism of CEUS in diagnosing blood perfusion of SIONFH.At the same time,observe the changes of the above observation parameters after taking compound Shengmaichenggu capsule,so as to provide a new detection method for the evaluation of Chinese medicine preparation.?based on clinical studies,to observe the characteristics of CEUS perfusion curve and the expression of quantitative parameters of femoral head in healthy adults and patients with SIONFH,compare the changes of CEUS of femoral head in necrosis group before and after taking compound Shengmaichenggu capsule,evaluate its therapeutic effect,and compare with the results of animal experiments in the first part to analyze the feasibility of CEUS in diagnosing ONFH.Overall,this study will preliminarily explore the sensitivity and specificity of CEUS in evaluating blood perfusion to provide a simple and effective test for early diagnosis and efficacy evaluation of clinical SIONFH,which is easy and convenient for follow-up observation.MethodsThis topic is conducted in two parts.Part I: 63 New Zealand rabbits were randomly divided into blank group(n=21)and model group(n=42)after quarantine qualifying.Rabbits in the model group received an ear edge injection of endotoxin combined with an intramuscular injection of methylprednisolone to prepare for ONFH model.After modeling,one rabbit in the blank group and two rabbits in the model group were randomly selected for histopathological examination every 1 week.The femoral head necrosis of rabbits in the model group was compared and observed.Three weeks after modeling,the femoral heads of two rabbits in the model group were positive for osteonecrosis.The rabbits in the model group were randomly divided into model group(n=18)and compound Shengmaichenggu capsule Chinese medicine group(n=18).From the third week,the Chinese medicine group was given Shengmaichenggu capsule suspension by gavage every day,and the model group and the blank group were given the same amount of normal saline once a day for 8 weeks.Six rabbits in each of the three groups were taken for CEUS micro blood perfusion examination of femoral head of both hind limbs at 5 weeks?7weeks and 11 weeks after modeling,and the venous blood of rabbit ear margin was collected for hypoxia inducible factor 1(HIF-1a)? The content of vascular endothelial growth factor(VEGF)was determined,and then all femoral heads were removed under sterile conditions for histopathological examination.Part II: A total of 30 patients with already MRI confirmed ARCO stage I-IIIA SIONFH were collected,including 40 femoral heads,and 10 healthy adult femoral heads were collected as normal control group.All patients with SIONFH were randomly divided into placebo control group and compound Shengmaichenggu capsule Chinese medicine group.Under the condition that the methods of supporting crutches,reducing weight and local physical therapy were consistent,the Chinese medicine group took compound Shengmaichenggu capsule 3 times/day,4 capsules/ time,and the control group took placebo with the same appearance for 6 months.CEUS was performed on all included femoral heads before and after treatment,the changes of femoral head micro blood flow perfusion before and after treatment were observed,and the curative effect of compound Shengmaichenggu capsule was evaluated.ResultsPart I: ? CEUS inspection: Two dimensional ultrasound observation showed that the femoral head of rabbits in the model group had smooth surface,no roughness and joint effusion,no obvious collapse in appearance,and there was no significant difference compared with the blank group.Quantitative analysis results: At 5 weeks,the ascending slope(as)and time to peak(TTP)of the curve in the model group were lower than those in the blank group,indicating that the overall flow rate and time of contrast medium on the surface of femoral head in the model group were accelerated and shortened.The descending slope(DS)and mean transit time(MTT)of the curve were lower than those in the blank group,indicating that the regression speed of the contrast agent in the overall microcirculation on the surface of the femoral head was accelerated,the effective perfusion time of the contrast agent in the femoral head was shortened,and the blood perfusion showed the performance of “fast forward and fast out”.There was no significant difference in CEUS parameters of femoral head at 7 and 11 weeks,P> 0.05.? Histopathological examination : Histopathological examination 2 weeks after modeling,2 cases of femoral head in the model group showed sparse and thin bone trabeculae and proliferation of bone marrow adipose tissue;Three weeks after modeling,all 4 cases of femoral head degenerated into osteonecrosis positive,which showed that the number of bone trabeculae decreased significantly,became thinner and broken,structural disorder,pyknosis and necrosis of bone nucleus,formation of a large number of empty bone lacunae,diffuse hemorrhage and necrosis of bone marrow,fat necrosis,local new bone tissue,and some femoral head necrosis were classified as grade II;Five weeks after modeling: in the model group,there were 12 cases of femoral head,7 cases were positive for osteonecrosis and 5cases were negative for osteonecrosis.Although there was no formation of a large number of empty bone lacunae and nuclear pyknosis was not obvious,the empty bone lacunae increased slightly,the formation of empty halo around the nucleus,the bone trabeculae became sparse and thin,and the growth of bone marrow adipose tissue was increased,of which 3 cases showed bone marrow adipose tissue hyperplasia or osteonuclear pyknosis in the subchondral area,And the number of empty bone lacunae increased slightly.After 7 weeks of modeling,12 cases of femoral head and 4 cases of histological examination in the model group had no abnormal changes;8 cases were negative for osteonecrosis.There were 12 cases of femoral head in the traditional Chinese medicine group,and no abnormal changes were found in histological examination.11 weeks after modeling: no abnormal changes were found in the femoral head of blank group,model group and traditional Chinese medicine group.? Micro vessel density(MVD)count of femoral head: After modeling for 5 weeks,compared with the blank group,MVD of femoral head in the model group decreased significantly(P<0.05),the number of micro vessels decreased,and MVD of traditional Chinese medicine group decreased slightly,but there was no significant difference(P>0.05);There was no significant difference in MVD of femoral head between groups at 7 and 11weeks(P>0.05).? Hematological indexes: After modeling for 5 and 7 weeks,compared with the blank group,the model group had HIF-1??VEGF level increased(P<0.05);Compared with the model group,HIF-1a in the Chinese medicine group expression level decreased slightly,the difference was not statistically significant(P>0.05),and the expression of VEGF decreased,the difference was statistically significant.Modeling 11 weeks,HIF-1?expression level continued to increase,and the difference was statistically significant(P<0.05).Compared with the model group,HIF-1a in the Chinese medicine group expression level decreased significantly(P<0.05),and there was no significant difference in VEGF among the three groups(P > 0.05).Part II:? Two dimensional ultrasound comparison: In normal health,the shape of the femoral head is normal,the articular surface is smooth,the shape of the femoral head in stage II necrosis is still normal,the articular surface is slightly rough,there is no collapse of the femoral head,the femoral head in stage III flattens or collapses to varying degrees,the continuity of some articular surfaces is interrupted,and the joint space is narrowed.? Comparison of CEUS quantitative parameters between groups: Compared with normal group and necrosis group,DS in necrosis group was lower than that in normal group,while PI,EI,E3,AS and AUC were higher than those in normal group(P<0.05);TTP,MTT,AS,DS and AUC in the Chinese medicine group decreased after treatment,which was statistically significant compared with that before treatment;There was no significant difference in the parameters of placebo group after treatment;After treatment,TTP and DS in the Chinese medicine group were higher than those in the placebo group,and MTT,AS and AUC were lower than those in the placebo group(P<0.05).Conclusion1.After histopathological verification,the rabbit SIONFH model was successfully established by injection of endotoxin + methylprednisolone.2.CEUS can quantitatively analyze the changes of microcirculation perfusion of SIONFH.With the increase of necrosis degree,the microcirculation perfusion of subchondral bone of femoral head is enhanced,showing a “fast forward and fast out” mode.CEUS quantitative parameters TTP,MTT,AS,DS and AUC can reflect the change trend of blood perfusion to a certain extent,which can indicate the severity of femoral head necrosis.3.The analytical expression of ONFH by CEUS quantitative parameters is consistent with the expression of histopathology,MVD count,hematological indexes HIF-1a and VEGF,which proves that CEUS can quantitatively analyze the degree of femoral head necrosis to a certain extent and provide effective diagnostic information for clinic.4.After 5 weeks of modeling,the blood HIF-1a of rabbits in Chinese medicine group levels were lower than those in the model group,suggesting that compound Shengmaichenggu capsule may regulate and improve HIF-1a in the hypoxic environment of the femoral head function of expression.After adult SIONFH was treated with compound Shengmaichenggu capsule,TTP and DS in the traditional Chinese medicine group were higher than those in the placebo group,and MTT,AS and AUC were lower than those in the placebo group,indicating that compound Shengmaichenggu capsule can effectively improve the micro blood flow perfusion in necrotic tissue.The animal experiment is consistent with the conclusions obtained in clinical research.5.CEUS can safely,noninvasively and intuitively evaluate the characteristics of micro blood flow perfusion in the femoral head,and can effectively evaluate the early hip preservation treatment in practical clinical application.Some quantitative parameters of CEUS are of certain value in the evaluation of the disease,guiding the treatment and curative effect evaluation. |
PDF Full Text Request