Objective To investigate the mechanism of Naringenin(NGN)in the treatment of steroid(glucosteroid)-induced osteonecrosis of the femoral head(SONFH).Methods A SONFH rat model was established using Methylprednisolone(MPS)treatment,followed by intervention with NGN and zinc protoporphyrin(ZnPP).Micro-CT was used to analyze the morphological changes in femoral head tissues,and the levels of osteocalcin(OCN)in rat serum as well as heme oxygenase-1(HO-1)and hypoxia-inducible factor-1α(HIF-1α)in femoral bone tissue were measured.A cellular model was constructed by treating MC3T3-E1 cells with Dexamethasone(DEX),followed by NGN intervention.Bioinformatics analysis combined with molecular docking technology was used to predict the target of NGN,and the Pulldown experiment was performed for validation.The expression of HO-1 was knocked down through cell transfection,to analyze the viability,proliferation,apoptosis,and migration of MC3T3-E1 cells,and angiogenesis assays were conducted to evaluate the angiogenic potential of human umbilical vein endothelial cells(HUVECs).Results Micro-CT analysis revealed that,compared with the control group,the trabecular thickness and trabecular number were significantly reduced in the MPS group,while the bone surface area/bone volume ratio and trabecular separation were significantly increased(P<0.001).In vitro experimental results indicated that DEX inhibited the proliferation of MC3T3-E1 cells,promoted cell apoptosis,and increased reactive oxygen species generation(P<0.01),and that DEX suppressed the formation of mineralized nodules,a key indicator of osteogenic differentiation,and downregulated the expression of osteogenesis-related genes(Runt-related transcription factor 2,osteopontin,osteocalcin)(P<0.01).However,NGN treatment partially reversed these effects.DEX significantly inhibited the migration of HUVECs,angiogenesis,and the expression of angiogenesis-related markers(platelet endothelial cell adhesion molecule-1,vascular endothelial growth factor,and von Willebrand factor)(P<0.01).In contrast,NGN treatment did not significantly affect the aforementioned effects,but the treatment with NGN conditioned medium[CM(NGN)]partially reversed these effects(P<0.01).Bioinformatics analysis combined with Pulldown assay results indicated that HO-1 was the target of NGN.DEX treatment significantly downregulated the expression of HO-1,while NGN intervention partially counteracted the inhibitory effect induced by DEX(P<0.01);knockdown of HO-1 negated the therapeutic effects of NGN(P<0.01).Compared with MPS administration alone,the combined administration of NGN and MPS upregulated the expression of HO-1 and HIF-1α in rat femoral head tissues.However,the HO-1 inhibitor ZnPP further upregulated the expression of HO-1 but downregulated the protein level of hypoxia-inducible factor-α(HIF-1α)(P<0.01).Conclusion NGN exerts its therapeutic effects on SONFH by activating the expression and activity of HO-1,which regulates the HIF-1α/VEGF pathway to promote osteoblast differentiation,bone formation,and angiogenesis.

Objective To investigate the mechanism of Naringenin(NGN)in the treatment of steroid(glucosteroid)-induced osteonecrosis of the femoral head(SONFH).Methods A SONFH rat model was established using Methylprednisolone(MPS)treatment,followed by intervention with NGN and zinc protoporphyrin(ZnPP).Micro-CT was used to analyze the morphological changes in femoral head tissues,and the levels of osteocalcin(OCN)in rat serum as well as heme oxygenase-1(HO-1)and hypoxia-inducible factor-1α(HIF-1α)in femoral bone tissue were measured.A cellular model was constructed by treating MC3T3-E1 cells with Dexamethasone(DEX),followed by NGN intervention.Bioinformatics analysis combined with molecular docking technology was used to predict the target of NGN,and the Pulldown experiment was performed for validation.The expression of HO-1 was knocked down through cell transfection,to analyze the viability,proliferation,apoptosis,and migration of MC3T3-E1 cells,and angiogenesis assays were conducted to evaluate the angiogenic potential of human umbilical vein endothelial cells(HUVECs).Results Micro-CT analysis revealed that,compared with the control group,the trabecular thickness and trabecular number were significantly reduced in the MPS group,while the bone surface area/bone volume ratio and trabecular separation were significantly increased(P<0.001).In vitro experimental results indicated that DEX inhibited the proliferation of MC3T3-E1 cells,promoted cell apoptosis,and increased reactive oxygen species generation(P<0.01),and that DEX suppressed the formation of mineralized nodules,a key indicator of osteogenic differentiation,and downregulated the expression of osteogenesis-related genes(Runt-related transcription factor 2,osteopontin,osteocalcin)(P<0.01).However,NGN treatment partially reversed these effects.DEX significantly inhibited the migration of HUVECs,angiogenesis,and the expression of angiogenesis-related markers(platelet endothelial cell adhesion molecule-1,vascular endothelial growth factor,and von Willebrand factor)(P<0.01).In contrast,NGN treatment did not significantly affect the aforementioned effects,but the treatment with NGN conditioned medium[CM(NGN)]partially reversed these effects(P<0.01).Bioinformatics analysis combined with Pulldown assay results indicated that HO-1 was the target of NGN.DEX treatment significantly downregulated the expression of HO-1,while NGN intervention partially counteracted the inhibitory effect induced by DEX(P<0.01);knockdown of HO-1 negated the therapeutic effects of NGN(P<0.01).Compared with MPS administration alone,the combined administration of NGN and MPS upregulated the expression of HO-1 and HIF-1α in rat femoral head tissues.However,the HO-1 inhibitor ZnPP further upregulated the expression of HO-1 but downregulated the protein level of hypoxia-inducible factor-α(HIF-1α)(P<0.01).Conclusion NGN exerts its therapeutic effects on SONFH by activating the expression and activity of HO-1,which regulates the HIF-1α/VEGF pathway to promote osteoblast differentiation,bone formation,and angiogenesis.

Objective To investigate the clinical value of visceral adipose tissue-derived serine protease inhibitor(vaspin)level in the diagnosis of nontraumatic osteonecrosis of femoral head(NONFH).Methods A total of 77 NONFH patients hospitalized in Linyi People's Hospital from November 2022 to May 2023 were included in necrotic group,and 76 healthy persons in our hospital during the same period were included in normal group.The general data of two groups were compared,and serum vaspin levels of patients with different clinical characteristics in necrotic group were compared.Receiver operating characteristic(ROC)curve was drawn to analyze the diagnostic value of serum vaspin in NONFH.Results The serum vaspin level in necrotic group was significantly lower than that in normal group(P<0.05).Serum vaspin levels in patients with bilateral hip disease were significantly lower than those in patients with unilateral hip disease(P<0.05).Serum vaspin levels in patients with femoral head collapse was significantly lower than that in patients without femoral head collapse(P<0.05).The serum vaspin level decreased with the increase of Association Research Circulation Osseous(ARCO)stage(P<0.05).Serum vaspin level was negatively correlated with ARCO stage and visual analogue scale score(P<0.05),but positively correlated with Harris hip score(P<0.05).ROC curve showed that the area under the curve of serum vaspin for NONFH was 0.868(95%CI:0.813-0.923,P<0.001),sensitivity was 82.9%,specificity was 75.3%.Conclusion Vaspin can be used as a potential serum marker for the diagnosis of NONFH.With the progression of NONFH,the serum vaspin level tends to decrease.

The pelvic incidence (PI) is a key anatomical parameter for vertebral-pelvic sagittal plane stabilization, that is, the angle between the perpendicular to the sacral plate at its midpoint and the line connecting this point to the middle axis of the femoral head, which fully reflects the compensatory ability of the pelvis and it does not change with age, gender and position, but is closely related to race. The measurement result of PI varies with the measurement method. The measurement on the standing lumbosacral lateral radiographs is a classic measurement method, which is easy to operate and has strong practicability. Later, the improved three-dimensional CT measurement and measurement methods on bone specimens are based on this, which reduces the error in X-ray measurement. Early research focused on PI and spine-related diseases. Compared with normal people, a small PI value is likely to cause intervertebral disc degeneration or even herniation, a high PI can cause lumbar spondylolisthesis. The measurement of PI can predict the risk of disc degeneration and lumbar spondylolisthesis progression, providing an important reference for spinal sagittal orthopedic reconstruction. The influence of PI in hip joint diseases cannot be ignored. PI also has a predictive effect on the development of certain hip joint diseases, and has guiding significance for the treatment of diseases. The relationship between PI and hip osteoarthritis, hip dysplasia, femur acetabular impingement, and non-traumatic necrosis of the femoral head was reviewed through literature review. The results showed that the relationship between hip osteoarthritis and PI was still controversial. The higher pelvic incidence was observed in Crowe I patients with hip dysplasia, and the lower PI was observed in patients with femur acetabular impingement. The higher PI was associated with a greater likelihood of femoral head collapse in patients with non-traumatic necrosis of the femoral head.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) as common seed cells have been widely used in tissue-engineered cartilage repair.OBJECTIVE: To use human amniotic membrane as a cell scaffold to carry rabbit BMSCs in order to repair articular cartilage defects in the femoral intercondylar fossa of rabbits.METHODS: Rabbit BMSCs were inoculated onto the human acellular amniotic membrane (HAAM) and co-cultured for 2 weeks. Articular defect models were made in the femoral intercondylar fossa of rabbits. The defects of the right knees served as blank control. BMSCs/HAAM composite was transplanted into the defect of the left knee joint as composite group, and HAAM was implanted into the defect of the left knee joint as HAAM group. These rabbits were killed at 8 and 12 weeks after implantation and the newly cartilage samples were evaluated grossly and histologically and then graded.RESULTS AND CONCLUSION: Gross observation showed the defects were filled with cartilaginous tissues in the composite group, and there were no cartilage tissues in the HAAM group, while only fibrous tissues were seen in the blank control group. Histologically, the defect region was full of chondrocytes in the composite group,immunohistochemistry staining indicated that collagen II was rich in the tissue, and furthermore, the cartilage matrix was stained deeply by toluidine blue. In the the HAAM group, there were few chondrocytes, toluidine blue staining was weakly positive, and immunohistochemistry staining was negative, indicating there was no cartilage matrix. In the blank control group, the defects were filled of fibroblasts and toluidine blue staining was weakly positive. To conclude, the BMSCs/HAAM is a good scaffold for BMSCs chondrogenic differentiation to effectively repair articular cartilage defects.

Objective To evaluate the effect of the bone grafting through a femoral neck window at high location in the treatment of osteonecrosis of the femoral head (ONFH).Methods From January 2009 to December 2011,26 patient (35 hips,11 males and 15 females) with ONFH were treated with bone grafting through fenestration at high level of femoral head and neck,and then they were followed up for average of 24 months,with the mean age of 37.4 years (range,18-65 years).According to the Association Research Circulation Osseous (ARCO) classification,there were 6 hips,13 hips,and 16 hips on stage Ⅱ A,Ⅱ B,Ⅱ C respectively.The patients were treated with the bone grafting through fenestration at high level of femoral head and neck.The fenestrated area was located in the junction of the femoral head and neck,involved part of the femoral head cartilage.The effectiveness was evaluated according to Harris Scoring system and excellent and good rate.Results All the cases were followed up in outpatient department for 24 months,and 1 was lost to follow-up.After the treatments with the bone grafting through fenestration at high level of femoral head and neck,no infections,nerve damage and other complications were found in all patients.Harris score of overall patients before and after treatment increased from 73.42±7.30 points to 85.85±11.63 points; Harris score of patients with ARCO Ⅱ A before and after treatment increased from 74.50± 1.76 points to 91.17± 1.60 points; Harris score of patients with ARCO Ⅱ B before and after treatment the average score increased from 73.92±8.03 points to 86.00±10.49 points.Harris score of patients with ARCO Ⅱ C before and after treatment increased from 72.60±8.29 points to 83.60± 14.29 points.The overall excellent and good rate before and after treatment in patients with preoperative 36.4％ up to 84.9％.The rate of patients with ARCO Ⅱ A increased from 33.3％ to 100.0％ after treatment.The rate of patients with ARCO Ⅱ B increased from 41.7％ to 91.7％ after treatment.The rate of patients with ARCO Ⅱ C increased from 33.3％ to 73.3％ after treatment.The difference in Harris scores and excellent and good rate between untreatment and treatment were statistical significant.Conclusion Bone grafting through a femoral neck window at high location in the treatment of osteonecrosis of the femoral head were proved to have good short-terms effects which can promote the restoration of osteonecrosis and improve the clinical symptoms of hip.

BACKGROUND: Recent studies have shown that steroid-induced osteonecrosis of the femoral head (ONFH) is associated with hereditary susceptibility. OBJECTIVE: To investigate the relationship between single-nucleotide polymorphisms and syndrome types in steroid-induced ONFH in populations from Linyi city, China. METHODS: The total dose of steroid, and the genetic polymorphisms of ApoB C7623T, G12619A, CYP1A2 G2964A sites were determined by polymerase chain reaction-ligase detection reaction combined with the sequencing analysis in 63 cases with ONFH by consecutive steroid therapy and 71 controlled cases without ONFH. RESULTS AND CONCLUSION: There were no significant differences in the total dose of steroid between two groups. The frequencies of ApoB C7623T TT genotype and CYP1A2 G2964A AG genotype in patients with ONFH were significantly higher than those without ONFH (P < 0.05). These experimental findings indicate that, ApoB C7623T and CYP1A2 G2964A polymorphisms may be associated to the susceptibility to steroid-induced ONFH, and the synergetic effect of these polymorphisms contributes to the onset of ONFH.