Osteoclast-mediated pain and collapse mechanisms in the femoral head necrosis with bone marrow edema / 西安交通大学学报(医学版)

【Objective】 To explore pain and collapse mechanisms in fosteonecrosis of the femoral head (ONFH) with bone marrow edema (BME). 【Methods】 ONFH patients at ARCO Ⅲ stage who underwent total hip arthroplasty in the First Affiliated Hospital of Guangzhou University of Chinese Medicine were enrolled; the femoral head samples, clinical and imaging data were collected. These patients were divided into BME group and non-BME group according to the MR data in one week preoperative. Hematoxylin-eosin and Sirius red staining were performed to observe the morphological changes in bone tissue of femoral head specimens. Western blotting and qPCR were used to semi-quantitatively analyze the expression levels of CTSK, RANKL, and Netrin-1 proteins and mRNA in different regions of the bone tissue. 【Results】 Clinical and imaging data showed that ONFH patients with BME had significantly higher scores of VAS than ONFH patients without BME. Hematoxylin-eosin staining showed that bone structure disorder and a large number of empty bone lacunae were found in the necrotic areas in both groups, but there exited significant granulation tissue in the BME group, and spindle-shaped fibroblastic cells and inflammatory cells aggregated in the repaired region. Sirius red staining revealed the necrotic and sclerotic areas were accumulated with many collagenous fiber in the BME group. The results of Western blotting and qPCR showed that Netrin-1 expressions in the necrotic, sclerotic and health areas in the BME group were higher than those in the non-BME group (P<0.05), while osteoclast related proteins and mRNA expressions of the necrotic and sclerotic areas in the BME group was higher than those in the non-BME group (P<0.05). 【Conclusion】 All these findings indicated that hip pain was positively correlated with femoral head necrosis with BME, hyperactive osteoclasts participated in the femoral head collapse with BME, and the upregulated expression of Netrin-1 mediated the pain mechanism.

A deep transfer learning method using plain radiographs for the differential diagnosis of osteonecrosis of the femoral head with other hip diseases / 中华骨科杂志

Objective:To develop a deep transfer learning method for the differential diagnosis of osteonecrosis of the femoral head (ONFH) with other common hip diseases using anteroposterior hip radiographs.Methods:Patients suffering from ONFH, DDH, and other hip diseases including primary hip osteoarthritis, non-infectious inflammatory hip disease, and femoral neck fracture treated in the First Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2018 to December 2020 were enrolled in the study. A clinical data set containing anteroposterior hip radiographs of the eligible patients was created. Data augmentation by rotating and flipping images was performed to enlarge the data set, then the data set was divided equally into a training data set and a testing data set. The ResNet-152, a deep neural network model, was used in the study, but the original Batch Normalization was replaced with Transferable Normalization to construct a novel deep transfer learning model. The model was trained to distinguish ONFH and DDH from other common hip diseases using anteroposterior hip radiographs on the training data set and its classification performance was evaluated on the testing data set.Results:The clinical data set was comprised of anteroposterior hip radiographs of 1024 hips, including 542 with ONFH, 296 with DDH, and 186 with other common hip diseases (56 hips with primary osteoarthritis, 85 hips with non-infectious inflammatory osteoarthritis, 45 hips with femoral neck fracture). After data augmentation, the size of the data set multiplied to 6144. The model was trained 100 050 times in each task. Accuracy was used as the representative parameter to evaluate the performance of the model. In the binary classification task to identify ONFH, the best accuracy was 95.80%. As for the multi-classification task for classification of ONFH and DDH from other hip diseases, the best accuracy was 91.40%. The plateau of the model was observed in each task after 50 000 times of training. The mean accuracy in plateaus was 95.35% (95% CI: 95.33%, 95.37%), and 90.85% (95% CI: 90.82%, 90.87%), respectively. Conclusion:The present study proves the encouraging performance of a deep transfer learning method for the first-visit classification of ONFH, DDH, and other hip diseases using the convenient and economical anteroposterior hip radiographs.

Role of bone status in anterolateral portion of femoral head in the progression of osteonecrosis of the femoral head / 中国组织工程研究

BACKGROUND:Mechanical stress plays an important role in the progressive collapse in osteonecrosis of the femoral head.Therefore,the bone structure of weight-bearing area may be the risk factor for collapse.OBJECTIVE:To investigate the relationship between the bone preservation of weight-bearing area and collapse progression in necrotic femoral head.METHODS:Eighty-seven patients (102 hips) with ARCO stage Ⅱ osteonecrosis of the femoral head,diagnosed by MRI,were enrolled.All patients underwent a natural progression.Collapse and bone involvement of the weight-bearing area were viewed by anteroposterior and frog-leg lateral radiographs.According to the location of the necrotic lesion on the anterolateral portion of the femoral head,the necrosis was divided into three types:type 1,the posteromedial and central portions;type 2,part of the anterolateral portion;type 3,the entire anterolateral portion.The collapse rate and the time to collapse in different types were assessed.RESULTS AND CONCLUSION:(1) All patients were followed up for 3-58 months.(2) Of the 60 hips with collapse,46 (76.7％) hip collapse was identified on anteroposterior radiograph.On frog-leg lateral radiograph showing collapsed femoral head could be identified in 57 (95.0％) hips,which was significantly different (P ＜ 0.01).(3) In all 102 hips,the collapse rate in type 3 osteonecrosis of the femoral head was significantly higher than that of type 2 osteonecrosis of the femoral head (P ＜ 0.001),and the time to collapse was markedly shortened.None collapse occurred in all six hips with type Ⅰ osteonecrosis of the femoral head during follow-up.(4) In summary,preservation of anterolateral portion is associated with potential collapse progression in necrotic femoral head.

Impact factors and clinical outcomes of the treatment for osteonecrosis of femoral head by using impacting bone graft: a long and mid-term study / 中华骨科杂志

Objective To investigate the long and mid-term clinical outcomes of the impacting bone graft and impact factors in treating osteonecrosis of femoral head (ONFH).Methods Impacting bone graft was utilized to treat 54 patients (64 hips)with ONFH.There were 44 cases male and 10 female with an average age of 35.44±8.86 (range,20-55) years.There were 5 hips caused by trauma,23 hips by glucocorticoid administration,28 hips by intake of alcohol,4 hips by alcohol and glucocorticoid induced,and 4 hips by idiopathic factors.There were 14 hips at ARCO (Association Research Circulation Osseous) stage Ⅱ,50 hips at stage Ⅲ and 6 hips at JIC (Japanese Osteonecrosis Investigation Committee) type B,23 hips at type C1,35 hips at C2 type.Harris hip score (HHS) was used to evaluate the clinical effects.Potential factors,including age,etiologies,pain duration,ARCO stage and JIC type,were evaluated to investigate their impacts on clinical outcomes.Surgery failure was defined as diverting to other surgeries,or poor HHS score evaluation (＜70 points).The survival time of femoral head was defined as the interval between time of impacting bone graft and the failure of surgery.Results The mean follow-up duration was 8.61 ± 1.45 (range,5.13-10.84) years.The proportion of patients with excellent or good Harris score was 81.3％ (52/64).The excellent or good rate was 76.0％ (38/50) at 8 years follow-up.Furthermore,the total survival rate of femoral head was 92％ at 8 years follow-up.The number of surgical failure was 10 (12 hips) and the failure rate was 18.8％ (12/64).There were no significant differences in ages,etiologies,pain duration,ARCO stage and JIC type among these patients before surgery (P ＜ 0.05).There were significant differences in the post-operational Harris score and excellent or good rate among the 20-30 years group,31-40 years group and over 40 years group (P ＜ 0.05).No significant difference for both two indexes was observed among different etiologies (P ＞ 0.05).When considering the duration after the initial pain,there were significant differences among 6 months group,7-12 months group and over 12 months group (P ＜0.05).There were significant differences between the Ⅲa stage and Ⅲc stage (P＜ 0.05),and between the C1 type and C2 type (P ＜ 0.05).Multifactor Logistic regression showed that there was closely relationship between the Harris score (excellent or good rate) after surgery and pain duration,ARCO stage,JIC type (P ＜ 0.05).Conclusion Impacting bone graft for ONFH is satisfy in the long and mid-term follow up duration.Impacting bone graft would be more suitable for patients with the collapse less than 2 mm,the lateral wall preservation and interval after the initial pain less than 12 months.

Flash Extraction Process Optimization of Chlorogenic Acid and Luteoloside from Lonicera Japonica by Box-Behnken Design and Response Surface Methodology / 中国药师

Objective: To optimize the flash extraction process of the active ingredients from Lonicera japonica by Box-Behnken design and response surface methodology.Methods: Flash extraction was used.With the concentration of ethanol,solid-liquid ratio and extraction time as the main influencing factors and the overall normalization value of the transfer rates of chorogenic acid and luteoloside as the evaluation index, Box-behnken design was performed to screen out the optimal extraction conditions.The mathematics relationship between the overall normalization value and the independent variables was established by multiple linear regression and binomial fitting, and response surface methodology was used to predict the optimal process conditions.Results: The optimal extraction conditions were determined as follows: the ethathol concentration was 66.92% ,the solid-liquid ratio was 18.82 ,the extraction duration was 1.20 min and extracted only once.Under the above conditions, the extraction rate of chloragenic acid and total luteoloside was 92.87% and 87.55% , respectively.Conclusion: The optimized technology is simple,economic and practical,which can provide new ideas for the rapid extraction and quality control of active ingredients from Lonicera japonica .

Expression of aromatase and estrogen-related receptors in human bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Estrogen signaling pathway for interaction between aromatase and estrogen-related receptor may exist in bone marrow mesenchymal stem cel s, which is used for regulating biological activity of bone marrow mesenchymal stem cel s. OBJECTIVE:To observe the expression of aromatase and estrogen-related receptors in adult bone marrow mesenchymal stem cel s during osteogenic differentiation. METHODS:Bone marrow mesenchymal stem cel s were respectively cultured in low-glucose DMEM medium (control group) and osteogenic induction medium (induction group). Cel proliferation and calcium deposition were determined by MTT assay and alizarin red staining, respectively. The expression of aromatase, estrogen receptorα, estrogen receptorβ, and estrogen-related receptorαduring osteogenic differentiation were determined by real-time PCR and western blot analysis. Estradiol levels in supernatants and lysates were detected by ELISA method. RESULTS AND CONCLUSION:In the induction group, the proliferation ability of bone marrow mesenchymal stem cel s was the strongest at 72 hours of culture;while there were a great amount of calcium nodules formed at 21 days of culture. Results from PCR and western blot assay showed that the expression of aromatase and estrogen receptorαwas improved in the induction group, but the expression of estrogen-related receptorαwas inhibited. There was no difference in the expression of estrogen receptorβbetween the two groups. ELISA results indicated that the level of estradiol in the supernatant of induction group was the highest. These findings indicate that aromatase, estrogen receptorα, estrogen receptorβand estrogen-related receptorαare al involved in osteogenesis of bone marrow mesenchymal stem cel s. Moreover, estradiol can be synthesized and secreted in bone marrow mesenchymal stem cel s, and most likely, promote the osteogenic differentiation of bone marrow mesenchymal stem cel s by related receptor pathway.

In vitro cultivation, identification and osteoinduction of adult bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cells have the potential of self-proliferation and multi-directional differentiation, while mesenchymal stem cells are few in adult bone marrow. In vitro purification, amplification and osteoinduction are very important for the research of bone tissue engineering. OBJECTIVE:To establish a simple and reliable in vitro cultivation and identification system of adult bone marrow mesenchymal stem cells, and to induce the mesenchymal stem cells to differentiate into osteoblasts. METHODS:Bone marrow were extracted from adult anterior superior iliac, the density gradient centrifugation and adhesion method were used to isolate, purify, culture and amplify the bone marrow mesenchymal stem cells. Osteogenic medium was prepared by mixing appropriate amount of dexamethasone,β-glycerophosphate and ascorbic acid C. The cells were divided into osteoinduction group and blank control group for observation. RESULTS AND CONCLUSION:Adult bone marrow mesenchymal stem cells were in typical long spindle-shape. The cells grew into rapid proliferation phase at 8-11 days and the growth curve was S-shape. CD44 and CD90 were in positive expression, while CD34 and CD45 were negative. The alkaline phosphatase activity was increased with culturing time prolonging, and reached the summit at the 12th day. The alkaline phosphatase activities of osteoinduction group were higher than those in the blank control group at different time points. These results suggested that in vitro cultivation, identification and osteoinduction system could obtain mesenchymal stem cells with high purity and good osteogenic differentiation capacity.

Study of model establishment of unicompartmental knee athroplasty based on three-dimensional parametric modeling method / 生物医学工程学杂志

The model of knee OA (KOA) was established in our laboratory with three-dimensional (3D) reconstruction technique based on the CT data. We then designed the unicompartmental prosthesis matching KOA model with parametric software Pro/e 5. 0. Finally, the models were used to mimic the operation of knee unicompantment replacement and the model of unicompartmental knee athroplasty (UKA) was analyzed using 3D finite element method. The results indicates that the approach for designing the model by 3D parametric is effective and meets the need of automation and efficacy to design unicompartmental prosthesis. The results proves that the model is vivid and precise, and the 3D parameteric modeling method can provide biomechanical research of unicompantment replacement with model foundation and new experimental method. It can be widely utilized in the clinics and research.