ObjectiveTo explore the effect of Kaempferol on bone microvascular endothelial cells (BMECs) in glucocorticoid induced osteonecrosis of the femoral head (GIONFH) in vitro. MethodsBMECs were isolated from cancellous bone of femoral head or femoral neck donated voluntarily by patients with femoral neck fracture. BMECs were identified by von Willebrand factor and CD31 immunofluorescence staining and tube formation assay. The cell counting kit 8 (CCK-8) assay was used to screen the optimal concentration and the time point of dexamethasone (Dex) to inhibit the cell activity and the optimal concentration of Kaempferol to improve the inhibition of Dex. Then the BMECs were divided into 4 groups, namely, the cell group (group A), the cells treated with optimal concentration of Dex group (group B), the cells treated with optimal concentration of Dex+1 μmol/L Kaempferol group (group C), and the cells treated with optimal concentration of Dex+5 μmol/L Kaempferol group (group D). EdU assay, in vitro tube formation assay, TUNEL staining assay, Annexin Ⅴ/propidium iodide (PI) staining assay, Transwell migration assay, scratch healing assay, and Western blot assay were used to detect the effect of Kaempferol on the proliferation, tube formation, apoptosis, migration, and protein expression of BMECs treated with Dex. ResultsThe cultured cells were identified as BMECs. CCK-8 assay showed that the optimal concentration and the time point of Dex to inhibit cell activity was 300 μmol/L for 24 hours, and the optimal concentration of Kaempferol to improve the inhibitory activity of Dex was 1 μmol/L. EdU and tube formation assays showed that the cell proliferation rate, tube length, and number of branch points were significantly lower in groups B-D than in group A, and in groups B and D than in group C (P<0.05). TUNEL and Annexin V/PI staining assays showed that the rates of TUNEL positive cells and apoptotic cells were significantly higher in groups B-D than in group A, and in groups B and D than in group C (P<0.05). Scratch healing assay and Transwell migration assay showed that the scratch healing rate and the number of migration cells were significantly lower in groups B-D than in group A, and in groups B and D than in group C (P<0.05). Western blot assay demonstrated that the relative expressions of Cleaved Caspase-3 and Bax proteins were significantly higher in groups B-D than in group A, and in groups B and D than in group C (P<0.05); the relative expressions of matrix metalloproteinase 2, Cyclin D1, Cyclin E1, VEGFA, and Bcl2 proteins were significantly lower in groups B-D than in group A, and in groups B and D than in group C (P<0.05). Conclusion Kaempferol can alleviate the damage and dysfunction of BMECs in GIONFH.
Objective To explore the impact of preoperative traction on the osteonecrosis of the femoral head (ONFH) in patients with femoral neck fractures. Methods Between February 2013 and May 2016, 120 patients with femoral neck fractures, who were treated with screw fixation, were collected. Sixty patients with fractures of Garden type Ⅰ and Ⅱ were non-displaced fracture group; 60 cases with fractures of Garden type Ⅲ and Ⅳ were displaced fracture group. The patients in 2 groups were randomly divided into traction and non-traction subgroups (n=30). There was no significant difference in gender, age, injury mechanism, damage side, the time from injury to operation, and fracture classification between 2 subgroups (P>0.05). Intracapsular pressure was recorded before operation. The quality of fracture reduction and the satisfaction ratio of screw implant were evaluated during operation. Visual analogue scale (VAS), Harris score, joint mobility, and the incidence of ONFH would be evaluated at 6 months, 1 year, and 2 years after operation. Results All incisions of 2 groups healed by first intention after operation. There was no infection or deep vein thrombosis of lower extremity. All patients were followed up 2 years. In displaced and non-displaced fracture groups, the intracapsular pressure of traction subgroups were higher than that of non-traction group (P<0.05); the differences of the quality of fracture reduction and the satisfaction ratio of screw implant were not significant (P>0.05) between 2 subgroups. At 6 months, 1 year, and 2 years after operation, VAS scores were higher in traction subgroup than in non-traction subgroup (P<0.05); and the joint mobility and Harris scores were lower in traction subgroup than in non-traction subgroup (P<0.05). X-ray films showed all fractures healed. Except for the non-displaced group at 6 months, the incidences of ONFH were higher in traction subgroup than in non-traction subgroup at other time points (P< 0.05). Conclusion Preoperative traction may increase the risk of ONFH, which can increase the intracapsular pressure and affect the blood supply of femoral head.
ObjectiveTo analyse the effectiveness of unicompartmental knee arthroplasty (UKA) for the patients with spontaneous osteonecrosis of the knee (SONK). MethodsBetween January 2012 and December 2016, 31 patients with SONK was admitted and treated with medial UKA. All patients were examined by both plain radiography and magnetic resonance images. The patients were composed of 5 men and 26 women with an average age of 64.3 years (range, 48-79 years), and with 16 left joints and 15 right joints. The average disease duration was 14.7 months (range, 6-26 months). Preoperative visual analogue scale (VAS) was 6.00±1.15, Hospital for Special Surgery (HSS) score was 55.77±11.03, and knee range of motion (ROM) was (114.68±10.40)°. The imaging examinations showed that all the lesions were located in the medial compartment of the knee joint and there were 19 patients with Aglietti stage Ⅳ and 12 patients with Aglietti stage Ⅴ. Preoperative femorotibial angle (FTA) was (177.39±1.63)° and posterior tibial slope (PTS) was (84.05±1.39)°. ResultsAll the incisions healed by first intention. All patients were followed up 14-46 months (mean, 25 months). At last follow-up, VAS score was 2.06±0.72 and HSS score was 86.45±3.67, which both improved significantly when compared with preoperative scores (t=22.73, P=0.00; t=–14.72, P=0.00). ROM was (118.06±3.80)° with no significant difference when compared with preoperative ROM (t=–1.78, P=0.08). The X-ray films showed there was no severe adverse events, such as periprosthetic infection, aseptic loosening, bearing dislocation, and so on. At last follow- up, PTS was (85.30±1.19)° with significant difference compared with preoperative one (t=–4.07, P=0.00); while FTA was (177.51±1.98)° with no significant difference when compared with preoperative FTA (t=–0.38, P=0.71). ConclusionUKA may be an optional management for SONK with minimally invasive, bone-preserving, and rapid recovery.
【Abstract】 Objective To approach the possibil ity of combination of simvastatin and BMSCs transplantation forsteroid-associated osteonecrosis of femoral head. Methods The BMSCs harvested from 24 rabbits were prepared for cell suspension at a concentration of 1 × 107/mL, and combined with gelatin sponge. Seventy New Zealand white rabbits received one intravenous injection of l ipopolysaccharide (10 μg/ kg). After 24 hours, three injections of 20 mg/kg of methylprednisolone were given intramuscularly at a time interval of 24 hours. Forty-eight rabbits diagnosed as having femoral head necrosis by MRI were divided into 4 groups randomly, group A: no treatment; group B: only decompression; group C: decompression and BMSCs transplantation; and group D: simvastatin drench (10 mg/kg.d) decompression and BMSCs transplantation. The general information of animals were recorded; after 4 and 8 weeks of operation, 6 rabbits of each group were chosen randomly to do MRI scan, and femoral heads were harvested to do histopathology and scanning electron microscope examination. Results After 8 weeks, rabbits became more active than before treatment, and walking way became normal gradually in groups C and D. Fourweeks after operation, the MRI low signal region of all groups had no obvious changes, but 8 weeks later, the necrosis signal region of group A magnified while it reduced obviously in group D. Histopathological observation: 4 weeks after operation, diffuse presence of empty lacunae and pyknotic nuclei of osteocytes were found in the trabeculae, and few newborn micrangium could been seen in group A; lots of empty lacunae and a small quantity of newborn micrangium could been found in group B; and large amounts of osteoblats and newborn micrangium were found around the necrosis regions in groups C and D. The positive ratio of empty lacunae and microvessel density in group D were 19.30 ± 1.52 and 7.08 ± 1.09, showing significant difference compared with other groups (P lt; 0.05). After 8 weeks of treatment, the bone trabecula collapsed in many regions in group A; there was fibra callus formation along the decompression channel in group B; few empty lacunae was in the bone trabecular, but the shape of marrow cavity was not normal in group C; and it showed almost normal appearance in group D. The positive ratio of empty lacunae and microvessel density in group D were 11.31 ± 1.28 and 12.37 ± 1.32, showing significant differences compared with other groups (P lt; 0.05), meanwhile, showing significant difference compared with that of 4 weeks after operation(P lt; 0.05). Scanning electron microscope: 8 weeks after operation, the bone trabecula collapsed in many regions, and few osteoblasts could be found on the surface, a great quantity of fat cells cumulated in the bone marrow in group A; cracked bone trabecula could be found occasionally in group B; the density of bone trabecula was lower than the normal in group C; and the shape of the marrow avity and thedensity of bone trabecula were similar to the normal in group D. Conclusion Simvastatin can promote the differentiation of osteocyte and vascular endothel ial cell from MSCs, the combination of simvastatin and marrow stem cells transplantation for the treatment of steroid-associated osteonecrosis of femoral head have good appl ication prospects.
Objective To evaluate the effects and the molecular mechanism of Liuwei dihuang pills in preventing steroid-induced osteonecrosis of the femoral head (ONFH) so as to provide an expremental basis for preventing ONFH cl inically. Methods Thirty-six adult Kunming mice (weighing 40-50 g, 46 g on average) were randomly divided into three groups (n=12): group A (control group), group B (model group) and group C (prevention group). In groups B and C, ONFHmice models were produced by intraperitoneal injection of horse serum at first (10 mL/kg) and a second injection of horse serum intraperitoneally (5 mL/kg) and prednisolone intramuscularly [45 mL/(kg•day), for 5 days] 2 weeks later. At the same time, the mice in group C were given Liuwei dihuang pills intragastrically [2 g/(kg•day)] and were given normal sal ine [10 mL/(kg•day)] in group B. In group A, mice were given normal sal ine intramuscularly and intragastrically as controls. The animals were sacrificed 2, 4, and 8 weeks after first treatment with prednisone, and femoral heads and l ivers were harvested to do histopathology analysis and apoptosis assay. Results Other mice survived throughout the experiment period except two death at 7 and 11 days after second injection of horse serum intraperitoneally in group B and one death at 24 hours after second injection of horse serum intraperitoneally in group C. The appearance and shape of the femoral head and the surface of cartilages were all normal. The histological observation showed: normal structures of l iver and femoral head were seen in group A at each time point; swell ing l iver cells with small fat vacuole, unclear structure of hepatic cords and narrower sinus hepaticus were seen, the bone trabeculae of femoral head was thin, sparse and collapsed in some regions and the changes became more obvious with time in group B; group C had similar results to group A. The percentage of empty osteocyte lacunae was significantly higher in group B than in groups A and C (P lt; 0.01). The osteoprotegrin expression significantly decreased and the osteoprotegrin l igand expressionsignificantly increased in group B when compared with groups A and C (P lt; 0.01). Apoptosis analysis showed that the apoptosis index in group B was significantly higher than that in groups A and C (P lt; 0.01). Conclusion Liuwei dihuang pills can prevent steroid-induced ONFH by improving l ipid metabol ism, releiving bone lesion, and protecting against cell death.
ObjectiveTo evaluate the short-term effectiveness of arthroscopic surgery combined with direct anterior approach for hip diseases.MethodsA retrospective study was performed on 23 cases with hip diseases (23 hips), who were treated with the arthroscopic surgery combined with direct anterior approach, between January 2015 and December 2016. There were 9 males and 14 females, aged from 27 to 49 years (mean, 38.6 years). There were 11 cases of posterior dislocation of the hip associated with femoral head fracture (Pipkin typeⅠ) and 7 cases of femoral neck fracture (Garden type Ⅳ). And the interval between injury and operation was 2-8 days (mean, 4.3 days). Five cases were osteonecrosis of femoral head at precollapse stage which were rated as stageⅡA according to Association Research Circulation Osseous (ARCO) classification system. The disease duration was 3-8 months (mean, 5.9 months). The preoperative Harris hip score, Oxford Hip Score (OHS), Postel score, and visual analogue scale (VAS) were 57.3±8.2, 11.2±3.6, 3.2±1.5, and 7.2±1.3, respectively.ResultsAll the wounds healed primarily. Lateral femoral nerve injury occurred in 3 cases. All patients were followed up 8-19 months (mean, 15.6 months). Bone union achieved in all patients after 14-19 weeks (mean, 15.8 weeks) and no secondary osteoarthritis or heterotopic ossification occurred. At last follow-up, the Harris hip score (92.5±5.3), OHS (36.5±5.9), and Postel score (14.2±2.6) were significantly higher than preoperative scores (t=45.274, P=0.000; t=36.586, P=0.000; t=32.486, P=0.000), and VAS score (1.8±0.9) was significantly lower than preoperative score (t=21.314, P=0.000).ConclusionArthroscopic surgery combined with direct anterior approach for hip diseases can effectively relieve pain, improve hip function, and obtain the satisfactory short-term effectiveness.
Objective To summarize retrospectively the clinical technology of repairing osteonecrosis of femoral head (ONFH) by free vascularized fibular grafting (FVFG), and the value of modified instruments in operation. Methods Between March 2011 and January 2013, 35 patients with ONFH (47 hips) who underwent FVFG with modified instruments. There were 24 males (32 hips) and 11 females (15 hips), aged 34 years on average (range, 22-43 years). The unilateral hip was involved in 23 cases and the bilateral hips in 12 cases. The disease duration ranged from 5 to 9 months (mean, 7 months). Based on etiology, 25 hips were classified as alcohol ONFH, 12 hips as corticosteroids ONFH, 3 hips as trauma ONFH, and 7 hips as idiopathic ONFH. According to the Association Research Circulation Osseous(ARCO) stage, 3 hips were rated as stage I, 39 hips as stage II, and 5 hips as stage III on the X-ray films. The preoperative Harris score was 58.2±6.1. Results The time to get fibula was 15-35 minutes (mean, 25 minutes). The operation time was 90-200 minutes (mean, 130 minutes), and the blood loss during operation was 150-500 mL (mean, 270 mL). All the patients achieved primary healing of incision, without complication of infection or deep vein thrombosis. All 35 patients were followed up 12-42 months, with an average of 28 months. The Harris score at final follow-up was 87.3±5.7, showing significant difference when compared with preoperative score (t=102.038,P=0.000). Radiographic results at final follow-up showed good position of fibula; and necrosis was improved in 9 hips, had no changes in 36 hips, and aggravated in 2 hips. Conclusion FVFG for ONFH can improve hip function effectively, and modified instruments can improve operation efficiency.
ObjectiveTo investigate whether exosomes derived from miR-27a-overexpressing human umbilical vein endothelial cells (HUVECs)—exo (miR-27a) can promote bone regeneration and improve glucocorticoids (GC) induced osteonecrosis of femoral head (ONFH) (GC-ONFH).MethodsThe exo (miR-27a) were intended to be constructed and identified by transmission electron microscopy, nanoparticle tracking analysis, Western blot, and real-time fluorescent quantitative PCR (qRT-PCR). qRT-PCR was used to evaluate the effect of exo (miR-27a) in delivering miR-27a to osteoblasts (MC3T3-E1 cells). Alkaline phosphatase staining, alizarin red staining, and qRT-PCR were used to evaluate its effect on MC3T3-E1 cells osteogenesis. Dual-luciferase reporter (DLRTM) assay was used to verify whether miR-27a targeting Dickkopf WNT signaling pathway inhibitor 2 (DKK2) was a potential mechanism, and the mechanism was further verified by qRT-PCR, Western blot, and alizarin red staining in MC3T3-E1 cells. Finally, the protective effect of exo (miR-27a) on ONFH was verified by the GC-ONFH model in Sprague Dawley (SD) rats.ResultsTransmission electron microscopy, nanoparticle tracking analysis, Western blot, and qRT-PCR detection showed that exo (miR-27a) was successfully constructed. exo (miR-27a) could effectively deliver miR-27a to MC3T3-E1 cells and enhance their osteogenic capacity. The detection of DLRTM showed that miR-27a promoted bone formation by directly targeting DDK2. Micro-CT and HE staining results of animal experiments showed that tail vein injection of exo (miR-27a) improved the osteonecrosis of SD rat GC-ONFH model.Conclusionexo (miR-27a) can promote bone regeneration and protect against GC-ONFH to some extent.
Extracorporeal shock wave (ESW), as a noninvasive, safe, and effective treatment, was applied to the treatment in osteonecrosis of femoral head (ONFH) since the end of last century. Although this therapy is more and more widely used, there are many traps and challenges. We recommend using the high-energy focus ESW to treat ONFH, rather than using the low energy radial pressure wave. Furthermore, for different types ONFH, scientific personalized treatment planning should be made first. There are demands of multicenter united researches for this unknown field of ONFH treated with ESW, and so to provide high-level evidence-based medicine evidence.
【Abstract】 Objective To explore the correlation between pain grading, stage of necrosis and bone marrow edema(BME) in nontraumatic osteonecrosis of femoral head (NONFH) so as to strengthen understandings about cl inical significance of BME in NONFH. Methods From October 2004 to October 2006, 97 patients (149 hips) with NONFH were treated. There were 68 males and 29 femals with an average age of 38.8 years (19-62 years). The disease course was from 20 days to 4 years. BME was identified grade 0 to grade 2 according to MRI. Based on grading scale of pain, pain grading were divided into no pain (grade 0), mild pain (grade 1) and moderate or severe pain (grade 2). According to Association Research Circulation Osseous staging system, NONFH were divided into I-IV stages. The incidence rate of BME in each pain grading and stages of necrosis was analyzed respectively. Contingency table analyses and rank sum tests were used to compare the difference of pain grading and stages of necrosis among these groups. Results The total incidence rate of BME was 73.15% (109/149), the incidence rateswere 84.38% in pain groups (108 /128) and 94.12% in the grade 2 (32/34). Pain grading correlated with BME rating (P lt; 0.001).The results of rank sum tests for several independent samples showed significant difference in BME among pain groups(P lt; 0.001). With the advance of pain scale, the mean rank of BME increased gradually(28.19 for grade 0, 78.94 for grade 1 and 96.12 for grade 2). BME was more commonly and clearly seen in stage Ⅱ（77.05%）and stage Ⅲ（82.81%）of NONFH. Stage I-III of NONFH correlated with BME rating (P lt; 0.001). The results of rank sum tests showed significant difference in BME rating among three stages (P lt; 0.001). With the advance of disease, the rank of BME rating increased gradually (39.07 for grade 0, 60.16 for grade 1 and 86.15 for grade 2 ). Conclusion BME is a sign that is accompanied with NONFH. The probabil ity and extent of BME correlated well with the pain and stage of NONFH．The condition of BME can be used as a index for the appraisal of advancement of disease and the judgment of treatment result.