Mechanical effect of changed femoral neck ante-version angles on the stability of an intertrochanteric fracture fixed with PFNA: A finite element analysis.

Objective: Change of femoral neck ante-version angle postoperatively due to inadequate reduction could result in unsatisfying treatment outcome of intertrochanteric fracture. However, the influence of increased or decreased femoral neck ante-version on the biomechanical stability of the bone-implant complex has rarely been studied. Methods: A finite element model of a complete normal human femur with normal femoral neck ante-version as 13° was established accurately by scanning a 64 year old female femur. The models of 31-A1.1 intertrochanteric fractures with different femoral neck ante-version angles of 3°, 5.5°, 8°, 10.5°, 13°, 15.5°, 18°, 20.5°, 23° were created. They were assembled with a proximal femoral nail anti-rotation (PFNA) device. The biomechanical differences with varying femoral neck ante-version angles were compared using finite element analysis method. Results: As the femoral neck ante-version angle gradually increased from 13° to 23°with a gradient of 2.5°, the peak von Mises stress was gradually increased from 137.82 MPa to 276.02 MPa. Similarly, the peak von Mises stress was gradually increased from 137.82 MPa to 360.12 MPa with the femoral neck ante-version angle decreased from 13° to 3°. When decreased ante-version angle of 7.5° and increased ante-version angle of 10° will exceed the yield strength of femoral (240.32 MPa), the risk of femoral fracture will increase significantly. The maximum displacement of the femur was significantly reduced for increased ante-version models than for decreased ante-version models, whether the changes of ante-version angles were 2.5°, 5°, 7.5° or 10°. The maximum stress of PFNA was found in the intersection of main nail and helical blade, and became greater gradually as the ante-version angle increased or decreased with a gradient of 2.5°. The maximum stress of PFNA was presented in the model 5.5° with the maximum stress of 724.42 MPa (near to the yield strength of titanium alloy of 700-1000 MPa), producing the breakage risk of PFNA. The maximum displacement of the PFNA was significantly reduced for increased ante-version models than for decreased ante-version models, whether the changes of ante-version angles were 2.5°, 5°, 7.5° or 10°. Conclusion: Based on the results of present study, it was demonstrated that the anatomical reduction of femoral neck ante-version was vital to secure the optimal stability. Abnormal femoral ante-version could increase the potential risk of failure for intertrochanteric fracture after PFNA. The stability of increased femoral ante-version (less than 10°) was superior to the stability of decreased ante-version (less than 5°) for the cases of difficulty to acquire anatomical reduction. The clinical implication of the finding was that increased femoral neck ante-version had an advantage of mechanical stability towards the decreased femoral neck ante-version for the cases of comminuted intertrochanteric fracture and failure of anatomical reduction.

Focal osteoporosis defect is associated with vertebral compression fracture prevalence in a bone mineral density-independent manner.

Introduction: Focal osteoporosis defect has shown a high association with the bone fragility and osteoporotic fracture prevalence. However, no routine computed tomography (CT)-based vertebral focal osteoporosis defect measurement and its association with vertebral compression fracture (VCF) were discussed yet. This study aimed to develop a routine CT-based measurement method for focal osteoporosis defect quantification, and to assess its association with the VCF prevalence. Materials and Methods: A total of 205 cases who underwent routine CT scanning, were retrospectively reviewed and enrolled into either the VCF or the control group. The focal bone mineral content loss (focal BMC loss), measured as the cumulated demineralization within bone void space, was proposed for focal osteoporosis defect quantification. Its scan-rescan reproducibility and its correlation with trabecular bone mineral density (BMD) and apparent microarchitecture parameters were evaluated. The association between focal BMC loss and the prevalence of VCF was studied by logistic regression. Results: The measurement of focal BMC loss showed high reproducibility (RMSSD = 0.011 mm, LSC = 0.030 mm, ICC = 0.97), and good correlation with focal bone volume fraction (r = 0.79, P < 0.001), trabecular bone separation (r = 0.76, P < 0.001), but poor correlation with trabecular BMD (r = 0.37, P < 0.001). The focal BMC loss was significantly higher in the fracture group than the control (1.03 ± 0.13 vs. 0.93 ± 0.11 mm; P < 0.001), and was associated with prevalent VCF (1.87, 95% CI = 1.31-2.65, P < 0.001) independent of trabecular BMD level. Discussion: As a surrogate measure of focal osteoporosis defect, focal BMC Loss independently associated with the VCF prevalence. It suggests that focal osteoporosis defect is a common manifestation that positively contributed to compression fracture risk and can be quantified with routine CT using focal BMC Loss.

Bioinformatics study of the potential therapeutic effects of ginsenoside Rf in reversing nonalcoholic fatty liver disease.

OBJECTIVE: Ginsenoside Rf, a tetracyclic triterpenoid only present in Panax ginseng, has been proven to relieve lipid metabolism and inflammatory reactions, which can be a potential treatment for nonalcoholic fatty liver disease (NAFLD). Therefore, this study aimed to reveal the underlying mechanisms of ginsenoside Rf in the treatment of early-stage NAFLD (NAFL) by using a bioinformatics method and biological experiments. METHODS: Target genes associated with NAFL were screened from the Gene Expression Omnibus (GEO) database, a database repository of high-throughput gene expression data and hybridization arrays, chips, and microarrays. Subsequently, gene set enrichment analysis was performed by using Gene Ontology enrichment analysis tool. Then, the binding capacity between ginsenoside Rf and NAFL-related targets was evaluated by molecular docking. Finally, the FFA-induced HepG2 cell model treated with ginsenoside Rf was adopted to verify the effect of ginsenoside Rf and the related mechanisms. RESULTS: There were 41 common differentially expressed genes in the GEO dataset. Gene Ontology and Reactome pathway enrichment analysis of the differentially expressed genes showed that many pathways could be related to the pathogenesis of NAFL, including those participating in the cytokine-mediated signaling pathway, G protein-coupled receptor signaling pathway, and response to lipopolysaccharide. Finally, the qRT-PCR analysis results indicated that ginsenoside Rf therapy could ameliorate the transcription of ANXA2, BAZ1A, DNMT3L and MMP9. CONCLUSION: Our research discovered the relevant mechanisms and basic pharmacological effects of ginsenoside Rf in the treatment of NAFL. These results might facilitate the development of ginsenoside Rf as an alternative medication for NAFL.

Finite element analysis of the kinematic coupling effect of the joints around talus when Ponseti manipulation.

BACKGROUND: Little information was obtained from the published papers about the kinematic coupling effect between tarsal bones during Ponseti manipulation. The aim was to explore the kinematic coupling effect of the joints around talus, to investigate the kinematic rhythm and coupling relationship of tarsal joints; to clarify the pulling effect on medial ligament of the ankle during the process of Ponseti manipulation. METHODS: The model of foot and ankle was reconstructed from the Chinese digital human girl No.1 (CDH-G1) image database. Finite element analysis was applied to explore the kinematic coupling effect of the joints around talus. The distal tibia and fibula bone and the head of talus were fixed in all six degrees of freedom; outward pressure was added to the first metatarsal head to simulate the Ponseti manipulation. Kinematic coupling of each tarsal joint was investigated using the method of whole model splitting, and medial ligament pulling of the ankle was studied by designing the model of medial ligament deletion during the Ponseti manipulation. RESULTS: All the tarsal joints produced significant displacement in kinematic coupling effect, and the talus itself produced great displacement in the joint of ankle. Quantitative analysis revealed that the maximum displacement was found in the joints of talonavicular (12.01mm), cuneonavicular (10.50mm), calcaneocuboid (7.97mm), and subtalar(6.99mm).The kinematic coupling rhythm between talus and navicular, talus and calcaneus, calcaneus and cuboid, navicular and cuneiform 1 were 1:12, 1:7, 1:2 and 1:1.6. The results of ligaments pulling showed that the maximum displacement was presented in the ligaments of tibionavicular (mean 27.99mm), talonavicular (21.03mm), and calcaneonavicular (19.18 mm). CONCLUSIONS: All the tarsal joints around talus were involved in the process of Ponseti manipulation, and the strongest kinematic coupling effect was found in the joints of talonavicular, subtalar, calcaneocuboid, and cuneonavicular. The ligaments of tibionavicular, talonavicular, and calcaneonavicular were stretched greatly. It was suggested that the method of Ponseti management was a complex deformity correction processes involved all the tarsal joints. The present study contributed to better understanding the principle of Ponseti manipulation and the pathoanatomy of clubfoot. Also, the importance of cuneonavicular joint should be stressed in clinical practice.

ß-Glucan from Saccharomyces cerevisiae alleviates oxidative stress in LPS-stimulated RAW264.7 cells via Dectin-1/Nrf2/HO-1 signaling pathway.

ß-Glucan from Saccharomyces cerevisiae has been described to be effective antioxidants, but the specific antioxidation mechanism of ß-glucan is unclear. The objectives of this research were to determine whether the ß-glucan from Saccharomyces cerevisiae could regulate oxidative stress through the Dectin-1/Nrf2/HO-1 signaling pathway in lipopolysaccharides (LPS)-stimulated RAW264.7 cells. In this study, we examined the effects of ß-glucan on the enzyme activity or production of oxidative stress indicators in LPS-stimulated RAW264.7 cells by biochemical analysis and the protein expression of key factors of Dectin-1/Nrf2/HO-1 signaling pathway by immunofluorescence and western blot. The biochemical analysis results showed that ß-glucan increased the LPS-induced downregulation of enzyme activity of intracellular heme oxygenase (HO), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) while decreasing the production of reactive oxygen species (ROS) and malondialdehyde (MDA). Furthermore, immunofluorescence results showed that ß-glucan can activate the nuclear factor erythroid 2-related factor 2 (Nrf2). The antioxidant mechanism study indicated that ß-glucan activated dendritic-cell-associated C-type lectin 1 (Dectin-1) receptors mediated Nrf2/HO-1 signaling pathway, thereby downregulating the production of ROS and thus produced the antioxidant effects in LPS-stimulated RAW 264.7 cells. In conclusion, these results indicate that ß-glucan potently alleviated oxidative stress via Dectin-1/Nrf2/HO-1 in LPS-stimulated RAW 264.7 cells.

Effect of Achilles tendon on kinematic coupling relationship between tarsal bones: a pilot finite element study.

BACKGROUND: The procedure of percutaneous Achilles tenotomy (PAT) is an important component of the Ponseti method. However, few studies reported the influence of Achilles tendon on kinematic coupling relationship between tarsal bones. The purpose of present study was to demonstrate the effect of Achilles tendon on the kinematic coupling relationship between tarsal bones, and to illustrate how kinematic coupling relationship between tarsal bones works in term of finite element analysis. METHODS: A three-dimensional finite element model of foot and ankle was constructed based on the Chinese digital human girl No.1 (CDH-G1) image database using the software of mimics, Geomagic studio, HyperMesh, and Abaqus. The last manipulation of the Ponseti method before the procedure of PAT was simulated. The talus head and the proximal tibia and fibula bone were fixed in all six degrees of freedom, and the outward pressure was added on the first metatarsal head to investigate the kinematic coupling relationship between tarsal bones. RESULTS: The least relationship of kinematic coupling between tarsal bones was found in calcaneus. Stress concentration was mainly observed at the navicular, talus and the medial malleolus. The difference in displacement of the navicular was only found with the Achilles tendon stiffness of 0 N/mm and others. No difference in the navicular displacement was found in the stiffness of Achilles tendon between 40, 80, 200, 400, and 1000 N/mm. The maximum displacement of navicular was observed at the ankle position of PF-20° (plantar flexion-20°). The difference in displacement of the navicular was greater at the ankle position of PF-20° with the Achilles tendon stiffness of 0 N/mm than that at the ankle position of PF-40° with the Achilles tendon stiffness of 40 N/mm. CONCLUSIONS: Based on the findings from this study, it was demonstrated that the Achilles tendon existence or not and ankle position had great influence, while increased stiffness of Achilles tendon had no influence on kinematic coupling relationship between tarsal bones. For the cases with severe equinus, earlier implementation of PAT procedure (with the purpose of release the Achilles tendon and reduce the degree of ankle plantar flexion) may be beneficial to the deformity correction.

[Directional breeding of high oil content peanut variety Yuhua 9 by in vitro mutagenesis and screening].

Leaf water potential of peanut subjected to drought stress is positively related to the oil content of peanut kernels. The aim of this study was to directly screen the high oil mutants of peanut and create the new peanut varieties using hydroxyproline as water potential regulator. In vitro mutagenesis was carried out with the embryonic leaflets of peanut variety Huayu 20 as explants and pingyangmycin as a mutagen added into the somatic embryo formation medium. The formed somatic embryos were successively transferred to somatic embryo germination and selection medium containing 6 mmol/L hydroxyproline (at -2.079 MPa water potential ) to induce regeneration and directionally screen high oil content mutants. After that, these plantlets were grafted and transplanted to the experimental field and 132 high oil mutants with oil content over 55% were obtained from the offspring of regenerated plants. Finally, among them, the oil contents of 27 lines were higher than 58% and of 2 lines were higher than 60%. A new peanut variety Yuhua 9 with high yield and oil content was bred from the regenerated plant progenies combining the pedigree breeding method. The yield was 14.0% higher than that of the control cultivar in the testing new peanut varieties of Liaoning province, and also it has passed the national registration of non-major crop varieties. Yuhua 9 with an oil content of 61.05%, which was 11.55 percentage points higher than that of the parent Huayu 20, was the peanut cultivar with the highest oil content in the world. The result showed that it was an effective way for directional breeding of high oil peanut varieties by means of the three-step technique including in vitro mutagenesis, directional screening by reducing water potential in medium and pedigree selection of regenerated plant progenies.

Controlled in vivo Bone Formation and Vascularization Using Ultrasound-Triggered Release of Recombinant Vascular Endothelial Growth Factor From Poly(D,L-lactic-co-glycolicacid) Microbubbles.

Bone defects are challenging to treat in musculoskeletal system due to the lack of vascularization. Biomaterials with internal vascularization ability and osteoinduction bioactivity are promising strategies for orthopedic applications. Vascular endothelial growth factor (VEGF) has been widely used for angiogenesis and osteogenesis. Here, we developed VEGF-loaded PLGA microbubbles (MBs) for improvement of angiogenesis and osteogenesis in bone defect repair in combination with ultrasound-targeted microbubble destruction (UTMD). Release profile showed UTMD promoted the burst release of VEGF from PLGA MBs. We subsequently investigated the combination of ultrasound application with VEGF MBs for in vitro osteogenesis. The results demonstrated that the expression of osteogenesis-related genes and calcium deposits were increased by VEGF MBs in combination of UTMD. Micro-computed tomography (micro-CT) and histological analysis were conducted 4 and 8 weeks post-surgery. In vivo results show that VEGF MBs in combination of UTMD could significantly enhance new bone formation and vascular ingrowth at the defect site in a rat calvarial defect model. In summary, VEGF MBs in combination of UTMD could augment bone regeneration and vascularization at calvarial bone defects and hold huge potential for clinical translation.

Inhibition of YAP with siRNA prevents cartilage degradation and ameliorates osteoarthritis development.

The Hippo/YAP signaling pathway is important for mediating organ size and tissue homeostasis, but its role in osteoarthritis (OA) remains unclear. We aimed to investigate the role of Hippo/YAP signaling pathway in OA development. YAP expression in OA cartilage was assessed by immunohistochemistry, RT-qPCR, and Western blotting. The effects of YAP overexpression or knockdown on gene expression related to chondrocyte hypertrophy induced by IL-1ß were examined. The in vivo effects of YAP inhibition were studied. Subchondral bone was analyzed by micro-CT. YAP was increased in mice and human OA articular cartilage and chondrocytes. YAP mRNA expression level was also increased in IL-1ß-induced chondrocytes. YAP overexpression resulted in increased expression of catabolic genes in response to IL-1ß. Suppression of YAP by siRNA inhibited IL-1ß stimulated catabolic genes expression and chondrocytes apoptosis. Intra-articular injection of YAP siRNA ameliorated OA development in mice. Micro-CT results showed the aberrant subchondral bone formation was also reduced. We provided evidence that YAP was upregulated in OA cartilage. Inhibition of YAP using YAP siRNA is a promising way to prevent cartilage degradation in OA. KEY MESSAGES: YAP was upregulated in human and mice osteoarthritis cartilage and chondrocytes. YAP siRNA decreased IL-1ß-induced catabolic gene expression. Intra-articular injection of YAP siRNA ameliorated OA development. Intra-articular injection of YAP siRNA reduced aberrant subchondral bone formation.

Timing for Ponseti clubfoot management: does the age matter? 90 children (131 feet) with a mean follow-up of 5 years.

Background and purpose - There are still controversies as to the age for beginning treatment with the Ponseti method. We evaluated the clinical outcome with different age at onset of Ponseti management for clubfoot. Patients and methods - 90 included children were divided into 3 groups in terms of age at start of treatment. The difference in treatment-related and prognosis-related variables including presentation age, initial Pirani and Dimeglio score, casts required, relapse rates, final Dimeglio score, and international clubfoot study group score (ICFSG) was analyzed. Results - Age between 28 days and 3 months at start of treatment method was associated with fewer casts required, lower relapse rate, and lower final ICFSG score (p < 0.05). Early treatment before 28 days of age required more casts and had a higher relapse rate (p < 0.05). The highest ICFSG scores were found in the ages between 3 and 6 months (p < 0.05). After propensity score matching, age between 28 days and 3 months was demonstrated to have a lower finial ICFSG score. Linear regression models showed that presentation age was positively correlated with final ICFSG score, and was identified as the only independent prognostic risk factor. Interpretation - There was lower rate of relapse and better clinical outcome when treatment was initiated at age between 28 days and 3 months. With the Ponseti method, clubfeet may not need urgent treatment.

Three-Dimensionally Printed Silk-Sericin-Based Hydrogel Scaffold: A Promising Visualized Dressing Material for Real-Time Monitoring of Wounds.

A wound dressing which can be convenient for real-time monitoring of wounds is particularly attractive and user-friendly. In this study, a nature-originated silk-sericin-based (SS-based) transparent hydrogel scaffold was prepared and evaluated for the visualization of wound care. The scaffold was fabricated from a hybrid interpenetrating-network (IPN) hydrogel composed of SS and methacrylic-anhydride-modified gelatin (GelMA) by 3D printing. The scaffold transformed into a highly transparent hydrogel upon swelling in PBS, and thus, anything underneath could be easily read. The scaffold had a high degree of swelling and presented a regularly macroporous structure with pores around 400 µm × 400 µm, which can help maintain the moist and apinoid environment for wound healing. Meanwhile, the scaffolds were conducive to adhesion and proliferation of L929 cells. A coculture of HaCaT and HSF cells on the scaffold showed centralized proliferation of the two cells in distributed layers, respectively, denoting a promising comfortable environment for re-epithelialization. Moreover, in vivo studies demonstrated that the scaffold showed no excessive inflammatory reaction. In short, this work presented an SS-based transparent hydrogel scaffold with steerable physical properties and excellent biocompatibility through 3D printing, pioneering promising applications in the visualization of wound care and drug delivery.

Using 3D finite element models verified the importance of callus material and microstructure in biomechanics restoration during bone defect repair.

BACKGROUND: There is lack of further observations on the microstructure and material property of callus during bone defect healing and the relationships between callus properties and the mechanical strength. METHODS: Femur bone defect model was created in rabbits and harvested CT data to reconstruct finite element models at 1 and 2 months. Three types of assumed finite element models were compared to study the callus properties, which assumed the material elastic property as heterogeneous (R-model), homogenous (H-model) or did not change from 1 to 2 months (U-model). RESULTS:  The apparent elastic moduli increased at 2 months (from 355.58 ± 132.67 to 1139.30 ± 967.43 MPa) in R-models. But there was no significant difference in apparent elastic moduli between R-models (355.58 ± 132.67 and 1139.30 ± 967.43 MPa) and H-models (344.79 ± 138.73 and 1001.52 ± 692.12 MPa) in 1 and 2 months. A significant difference of apparent elastic moduli was found between the R-model (1139.30 ± 967.43 MPa) and U-model group (207.15 ± 64.60 MPa) in 2 months. CONCLUSIONS: This study showed that the callus structure stability remodeled overtime to achieve a more effective structure, while the material quality of callus tissue is a very important factor for callus strength. At the meantime, this study showed an evidence that the material heterogeneity maybe not as important as it is in bone fracture model.

Long-term delivery of rhIGF-1 from biodegradable poly(lactic acid)/hydroxyapatite@Eudragit double-layer microspheres for prevention of bone loss and articular degeneration in C57BL/6 mice.

Insulin-like growth factor (IGF-1) has encouraged researchers to investigate its various potential therapeutic uses such as in the treatment of osteoporosis and repair of articular cartilage. The purpose of this study was to develop core-shell double-layer structural microspheres for the long-term delivery of rhIGF-1 for the prevention of bone loss and articular degeneration. rhIGF-1-loaded poly(lactic acid)/hydroxyapatite@Eudragit (PLA/HA@Eu) double-layer microspheres with a core-shell structure were formulated using poly(lactic acid) and hydroxyapatite by an oil-in-water-in-oil (w/o/w) technique. The in vitro release profiles of rhIGF-1 and the surface morphology and cytocompatibility of microspheres were investigated. In vitro release of rhIGF-1 from PLA/HA@Eu microspheres was maintained over 180 days. In vivo experiments were performed on male C57BL/6 mice with two different ages: adult mice (three months old) and old mice (ten months old). PLA/HA@Eu microspheres containing 1 mg rhIGF-1 were subcutaneously implanted into the back of mice. The rhIGF-1 amounts in the sera, livers and bones of mice were monitored for 6 months. The in vivo rhIGF-1 release patterns were similar to those observed from in vitro release. By analyzing the effects of rhIGF-1-loaded microspheres on bone mass in adult mice and cartilage in old mice via micro-CT and histological measurements, we observed beneficial trends. These results suggest that PLA/HA@Eu double-layer microspheres providing long-term delivery of rhIGF-1 were effective in systemic preventive therapy of bone loss in adult mice and delaying the progressive degeneration of articular cartilage in old mice. The long-term delivery strategy demonstrated herein may be a promising therapeutic approach for delaying osteoporosis and articular degeneration.

Notch signaling represses hypoxia-inducible factor-1α-induced activation of Wnt/ß-catenin signaling in osteoblasts under cobalt-mimicked hypoxia.

The modification of Wnt and Notch signaling pathways by hypoxia, and its association with osteoblast proliferation and apoptosis remain to be fully elucidated. To investigate Wnt-Notch crosstalk, and its role in hypoxia-induced osteoblast proliferation and apoptosis regulation, the present study investigated the effects of cobalt­mimicked hypoxia on the mouse pre-osteoblast-like cell line, MC3T3­E1, when the Notch signals were repressed using a γ­secretase inhibitor DAPT. The data showed that the cobalt­mimicked hypoxia suppressed cell proliferation under normal conditions, but increased cell proliferation under conditions of Notch repression, in a concentration­dependent manner. The results of western blot and reverse transcription­quantitative polymerase chain reaction analyses showed that the cobalt treatment increased the levels of activated ß­catenin protein and the expression levels of the target genes, axis inhibition protein 2 and myelocytomatosis oncogene, under DAPT­induced Notch repression. However, no significant changes were found in the expression levels of the Notch intracellular domain protein or the Notch target gene, hes1. In a ß­catenin gene­knockdown experiment, the proliferation of the MC3T3­E1 cells under hypoxia were decreased by DAPT treatment, and knockdown of the expression of hypoxia­inducible factor­1α (HIF­1α) suppressed the cobalt­induced increase in Wnt target gene levels. No significant difference in cell proliferation rate was found following DAPT treatment when the expression of HIF­1α was knocked down. The results of the present study showed the opposing effects of Wnt and Notch signaling under cobalt­mimicked hypoxia, which were partially regulated by HIF­1α, The results also showed that osteoblast proliferation was dependent on Wnt-Notch signal crosstalk.