LRP1 in GABAergic neurons is a key link between obesity and memory function.

OBJECTIVE: Low-density lipoprotein receptor-related protein-1 (LRP1) regulates energy homeostasis, blood-brain barrier integrity, and metabolic signaling in the brain. Deficiency of LRP1 in inhibitory gamma-aminobutyric acid (GABA)ergic neurons causes severe obesity in mice. However, the impact of LRP1 in inhibitory neurons on memory function and cognition in the context of obesity is poorly understood. METHODS: Mice lacking LRP1 in GABAergic neurons (Vgat-Cre; LRP1loxP/loxP) underwent behavioral tests for locomotor activity and motor coordination, short/long-term and spatial memory, and fear learning/memory. This study evaluated the relationships between behavior and metabolic risk factors and followed the mice at 16 and 32 weeks of age. RESULTS: Deletion of LRP1 in GABAergic neurons caused a significant impairment in memory function in 32-week-old mice. In the spatial Y-maze test, Vgat-Cre; LRP1loxP/loxP mice exhibited decreased travel distance and duration in the novel arm compared with controls (LRP1loxP/loxP mice). In addition, GABAergic neuron-specific LRP1-deficient mice showed a diminished capacity for performing learning and memory tasks during the water T-maze test. Moreover, reduced freezing time was observed in these mice during the contextual and cued fear conditioning tests. These effects were accompanied by increased neuronal necrosis and satellitosis in the hippocampus. Importantly, the distance and duration in the novel arm, as well as the performance of the reversal water T-maze test, negatively correlated with metabolic risk parameters, including body weight, serum leptin, insulin, and apolipoprotein J. However, in 16-week-old Vgat-Cre; LRP1loxP/loxP mice, there were no differences in the behavioral tests or correlations between metabolic parameters and cognition. CONCLUSIONS: Our findings demonstrate that LRP1 from GABAergic neurons is important in regulating normal learning and memory. Metabolically, obesity caused by GABAergic LRP1 deletion negatively regulates memory and cognitive function in an age-dependent manner. Thus, LRP1 in GABAergic neurons may play a crucial role in maintaining normal excitatory/inhibitory balance, impacting memory function, and reinforcing the potential importance of LRP1 in neural system integrity.

Incidence and outcomes of intraoperative periprosthetic acetabular fractures during cementless total hip arthroplasty: a prospective three-dimensional computer tomography-based study.

PURPOSE: Unlike periprosthetic femoral fractures, periprosthetic acetabular fractures during total hip arthroplasty (THA) have not been evaluated in detail. We prospectively evaluated the incidence, patterns, risk factors, and clinical outcomes of intraoperative periprosthetic acetabular fractures using pre- and postoperative computer tomography (CT). METHODS: In this prospective single-centre study, we evaluated 234 consecutive patients (250 hips) who underwent THA and three-dimensional CT before and after the surgery. We assessed the incidence, pattern of fractures, outcomes for each fracture pattern, reoperation and revision rates, Harris hip score, and visual analog scale (VAS) for pain. Multivariate regression models were used to identify risk factors for periprosthetic acetabular fractures. RESULTS: In total, 43 periprosthetic acetabular fractures (17.2%) were identified via CT. Fractures occurred most frequently at the superolateral wall. Early cup migration occurred in three hips. None of the patients underwent revision surgery for acetabular loosening. Regression modeling showed that rheumatoid arthritis was a significant predictor of periprosthetic acetabular fractures. CONCLUSIONS: Periprosthetic acetabular fractures are not infrequent during cementless THA and are more common in patients with rheumatoid arthritis.

Shape phenotype of thigh fat and muscle and risk of major adverse cardiovascular events after fragility hip fracture.

BACKGROUND: Although sarcopenia has been recognized as a predictor of mortality in elderly patients with hip fracture, the association of thigh fat and muscle with cardiovascular (CV) outcome remains unclear. We examined the impact of computed tomography (CT)-derived shape features of thigh fat and muscle on major adverse CV events (MACE) in elderly patients with hip fracture. METHODS: We conducted a retrospective analysis of consecutive patients aged ≥65 years who presented with hip fracture confirmed on pelvic bone CT scan and underwent hip fracture surgery at our institution from April 2019 to December 2021. The cross-sectional area (CSA) and compactness (CM) of both the muscle and fat at the upper-thigh level were calculated from two-dimensional CT images using AVIEW Research (v1.1.38, Coreline Soft, Co. Ltd, Seoul, South Korea). The shape features of thigh fat and muscle were categorized into four groups based on the combination of CSA and CM: fat CSA (fat area [FA])/fat CM (FCM), muscle CSA (muscle area [MA])/muscle CM (MCM), FA/MCM and MA/FCM. In each of them, subjects were categorized into four subgroups: high CSA/high CM, high CSA/low CM, low CSA/high CM and low CSA/low CM. The primary outcome was MACE after 30 days of surgery, defined as a composite of all-cause death, acute myocardial infarction, stroke or hospitalization for heart failure. RESULTS: Of 356 patients enrolled (median age, 82 years; 76.7% females), 72 (20.2%) had MACE over a median follow-up of 13.1 months (ranges 5.9-21.0 months). Patients with MACE had a significantly lower median FA (193.7 vs. 226.2 cm2 , P < 0.0001) and FCM (0.443 vs. 0.513, P = 0.001) compared with those without MACE, but no significant differences were found in MA, MCM and FA-MA ratio between the two groups. In a multivariate Cox regression analysis, low FA (<240.1 cm2 ) (adjusted hazard ratio [HR] 2.99, 95% confidence interval [CI] 1.39-6.44, P = 0.005) and low FCM (<0.477) (adjusted HR 2.00, 95% CI 1.10-3.63, P = 0.023) were associated with an increased risk of MACE. Among the shape phenotypes of thigh fat and muscle, the thigh fat phenotype of low FA/low FCM (adjusted HR 3.13, 95% CI 1.81-5.42, P < 0.0001 [reference, high FA/high FCM]) was found to be an independent predictor of MACE. CONCLUSIONS: In elderly patients with fragility hip fracture, thigh CT-derived measures of FA and FCM may provide useful prognostic information for predicting adverse CV outcomes.

Mimicking hypomethylation of FUS requires liquid-liquid phase separation to induce synaptic dysfunctions.

The hypomethylation of fused in sarcoma (FUS) in frontotemporal lobar degeneration promotes the formation of irreversible condensates of FUS. However, the mechanisms by which these hypomethylated FUS condensates cause neuronal dysfunction are unknown. Here we report that expression of FUS constructs mimicking hypomethylated FUS causes aberrant dendritic FUS condensates in CA1 neurons. These hypomethylated FUS condensates exhibit spontaneous, and activity induced movement within the dendrite. They impair excitatory synaptic transmission, postsynaptic density-95 expression, and dendritic spine plasticity. These neurophysiological defects are dependent upon both the dendritic localisation of the condensates, and their ability to undergo liquid-liquid phase separation. These results indicate that the irreversible liquid-liquid phase separation is a key component of hypomethylated FUS pathophysiology in sporadic FTLD, and this can cause synapse dysfunction in sporadic FTLD.

Correlation between Harris hip score and gait analysis through artificial intelligence pose estimation in patients after total hip arthroplasty.

BACKGROUND: Recently, open pose estimation using artificial intelligence (AI) has enabled the analysis of time series of human movements through digital video inputs. Analyzing a person's actual movement as a digitized image would give objectivity in evaluating a person's physical function. In the present study, we investigated the relationship of AI camera-based open pose estimation with Harris Hip Score (HHS) developed for patient-reported outcome (PRO) of hip joint function. METHOD: HHS evaluation and pose estimation using AI camera were performed for a total of 56 patients after total hip arthroplasty in Gyeongsang National University Hospital. Joint angles and gait parameters were analyzed by extracting joint points from time-series data of the patient's movements. A total of 65 parameters were from raw data of the lower extremity. Principal component analysis (PCA) was used to find main parameters. K-means cluster, X-squared test, Random forest, and mean decrease Gini (MDG) graph were also applied. RESULTS: The train model showed 75% prediction accuracy and the test model showed 81.8% reality prediction accuracy in Random forest. "Anklerang_max", "kneeankle_diff", and "anklerang_rl" showed the top 3 Gini importance score in the Mean Decrease Gini (MDG) graph. CONCLUSION: The present study shows that pose estimation data using AI camera is related to HHS by presenting associated gait parameters. In addition, our results suggest that ankle angle associated parameters could be key factors of gait analysis in patients who undergo total hip arthroplasty.

Accelerometer-derived physical activity analysis of elderly osteoarthritis patients.

BACKGROUND: Because disability in Osteoarthritis (OA) may change physical activity (PA), which might affect the disease progression, it is important to measure a patient's daily PA to study the relationship between a patient's PA and disease progression. OBJECTIVE: The objective of the present study was to investigate the relationship between PA and patients with OA and people without OA using data from the Korea National Health and Nutrition Examination Survey (KNHANES). METHODS: Demographic study was conducted to obtain data of comorbidities of participants. PA was compared between the group with OA (OA group) and the group without OA (non-OA group). In addition, PAs of OA patients with comorbidities and those without comorbidities were compared. The cut-off of moderate to vigorous physical activity (MVPA) was obtained through a receiver operating characteristic (ROC) curve. RESULTS: In the demographic study, there were significantly more educated participants in the OA group (p < .001). Actigraph data showed a significant decrease in MVPA (p < .001) but a significant increase in light activity (p = .002) in the OA group. In addition, the OA group showed significantly lower light PA but significantly higher MVPA in ≥10 min bout length. OA patients with comorbidities showed higher MVPA than OA patients without comorbidities (p = .044). The cut-off point of MVPA was 7.071 min/day when ROC curve was conducted. CONCLUSIONS: The present study suggests that patients with OA and low activity need a certain level of physical activity and a cut-off point for MVPA is presented which accounts for comorbidities in OA patients.

The incidence of focal osteonecrosis of the femoral head and its impact on conversion hip arthroplasty after screw fixation of femoral neck fractures: a multicenter retrospective study based on postoperative magnetic resonance imaging.

PURPOSE: Osteonecrosis of the femoral head (ONFH) and subsequent head collapse is a major concern after internal fixation of femoral neck fracture (FNF). Previous studies focused on ONFH incidence using plain radiography; postoperative magnetic resonance imaging (MRI) was rarely performed. We performed a multicenter retrospective study to investigate the incidence of ONFH and the need for conversion hip arthroplasty after FNF screw fixation. METHODS: We reviewed 195 patients who underwent screw fixation during closed FNF reduction between 2012 and 2017 at three institutions. Except for patients who did not consent to MRI, all patients underwent postoperative MRI either 1-3 years after screw fixation. The occurrence of ONFH was investigated through plain radiography and MRI. RESULTS: Thirty patients were diagnosed with ONFH through plain radiography, and an additional 33 patients were diagnosed with MRI, resulting in a total of 63 patients (32.3%) diagnosed with ONFH. The mean time to ONFH diagnosis was 18.9 months and the conversion rate to hip arthroplasty was 10.2%. Of the 33 patients who were normal on hip radiography but exhibited ONFH on MRI, all had small focal lesions not associated with head collapse at the last follow-up. The ONFH group diagnosed through plain radiography had more unstable FNFs than the group diagnosed through MRI. CONCLUSION: Although postoperative MRI revealed a higher incidence of ONFH after FNF screw fixation than reported previously, the small focal MRI lesions were not associated with increased risks of femoral head collapse or conversion to arthroplasty.

Caspase-dependent apoptosis induces reactivation and gliogenesis of astrocytes in adult mice.

Astrocytes play an important role in increasing synaptic plasticity, regulating endogenous homeostasis, and contributing to neuroprotection but become overactivated or apoptotic in persistent neuroinflammatory responses or pathological conditions. Although gliogenesis under these conditions may be essential for neuronal protection, much remains unknown. Here, we generated new conditional transgenic mice (cTg) that can induce apoptosis via Cre-dependent active caspase-3 (taCasp3-2A-TEVp) without pathological conditions. We induced apoptosis of hippocampal CA1 astrocytes in cTg mice using GFAP promoter-driven adeno-associated virus (AAV) containing Cre recombinase. Activated caspase-3 was detected in astrocytes of the hippocampal CA1, and the number of astrocytes decreased sharply at 1 week but recovered at 2 weeks and was maintained until 4 weeks. Nuclear factor 1A (NF1A) mRNA, an important transcription factor for hippocampal reactive astrocytes, was significantly increased only at week 1. Interestingly, all reactive markers (pan, A1, A2) increased despite the decreased number of astrocytes at week 1, and there was no change in monoamine oxidase B (MAOB) observed in astrocytes of animal models of degenerative brain disease. Extensive CA1 astrocyte depletion at week 1 induced cognitive deficits; however, both recovered at weeks 2 and 4. Overall, transient hippocampal astrocyte depletion caused by apoptosis restored cell number and function within 2 weeks and did not induce significant neurotoxicity. Therefore, cTg mice are valuable as an in vivo animal model for studying gliogenesis in multiple regions of the adult brain.

CJ14939, a Novel JAK Inhibitor, Increases Oxaliplatin-induced Cell Death Through JAK/STAT Pathway in Colorectal Cancer.

BACKGROUND/AIM: Colorectal cancer is reported to have the highest mortality rate among human malignancies. Although many research results for the treatment of colorectal cancer have been reported, there is no suitable treatment when resistance has developed. Therefore, it is necessary to develop new therapeutic agents. Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling plays an essential role in cell differentiation, proliferation, and survival. Abnormal activation of the JAK/STAT signaling pathway, by gene mutation or amplification, may induce cancer development, and sustained JAK/STAT activation is involved in chemoresistance. While many therapeutic agents have been developed to treat colon cancer, there remains no drug to overcome resistance to chemotherapies. The purpose of this study was to determine the potential of CJ14939 as a novel JAK inhibitor for the treatment of colorectal cancer. MATERIALS AND METHODS: In this study, cell culture, cell death assay, 3- (4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, colony formation assay, immunoblot analysis and tumor xenograft were applied. RESULTS: CJ14939 induced cell death, and inhibited phosphorylation of JAK1 and STAT3 in colorectal cancer cells. Furthermore, CJ14939 also promoted oxaliplatin-induced cell death, up-regulated expression of cleaved caspase-3, and down-regulated expression of phospho-JAK1 and phospho-STAT3. In vivo, co-treatment with CJ14939 and oxaliplatin notably reduced tumor growth when compared with CJ14939 or oxaliplatin treatment alone. CONCLUSION: This study identifies the important potential of CJ14939 in colorectal cancer treatment and suggests that combining CJ14939 with oxaliplatin might be a novel therapeutic strategy for patients with colorectal cancer.

Improved AAV vector system for cell-type-specific RNA interference.

BACKGROUND: RNA interference (RNAi) is a powerful technique to effectively silence or knock down gene function in mammalian cells. For better cell-type RNAi experiments in vivo, AAV vector-based RNA interference systems need to be improved. New method: In this study, we developed an AAV vector (CREon shRNA) that expressed CRE-dependent short hairpin RNA (shRNA) and fluorescent proteins simultaneously. RESULTS: We verified the Cre-dependent knockdown efficiency of the newly developed CREon shRNA vector in both HEK293T cells overexpressing TREK-1 and PC3 cells with endogenous TREK-1. Next, we packaged this TREK-1 CREon vector with AAV and injected it into the hippocampus of the brain together with a synapsin or GFAP promoter-driven CRE virus, confirming that it works well cell-selectively even in vivo. Finally, this viral vector was applied to an animal model of LPS-induced depression to determine whether behavioral changes occurred. Comparison with existing methods: With the existing pSico or pAAV-Sico-Red vectors, expression of fluorescent protein disappears when shRNA is conditionally activated by CRE recombinase, but our Creon shRNA vector showed simultaneous expression of both shRNA and fluorescent protein. Thus, it offers the advantage of allowing easy visual distinction of knocked-down cells. CONCLUSION: The newly improved CREon shRNA vector can be used as a novel research tool for conditional shRNA, and may be useful for various in vivo studies such as cancer and neurobiology.

Lateral pillar is the key in supporting pre-collapse osteonecrosis of the femoral head: a finite element model analysis of propensity-score matched cohorts.

BACKGROUND: This study was designed as a cohort study using propensity-score matching to age, gender, and body mass index (BMI) for finite element model (FEM) analysis from pre-collapse CT images of collapsed and non-collapsed hips. Through FEM analysis, a global graphical output around the hip joint can provide simple impression of stress distribution: concentration or dispersion. METHODS: A total of 32 hips with ARCO stage 2 or 3 ONFH who were on follow up for over a one-year period were retrospectively reviewed. 16 hips with no interval progression of collapse were set as the study group, then 16 hips with progression of collapse which required arthroplasty were set as the control group using propensity-score matching. FEM was generated through Mechanical Finder for each patient, then 4500 N of load was applied to 1000 mm2 area at the top of iliac crest to analyze the models in terms of equivalents for yield stress. RESULTS: Age, sex, and BMI had no significant differences between the two groups, while location (p = 0.015) was lateral, and size (p = 0.015) was significantly greater in the collapsed group. Non-collapsed hips mostly exhibited stress dispersion allocated to medial and lateral pillars, while collapsed hips exhibited stress concentration focused on the lateral pillar and the primary compression trabecula. (p = 0.001). CONCLUSION: Through FEM analysis, stress concentration to the lateral pillar and the primary compression trabeculae can be used to predict future collapse in ONFH with high probability. Results provide a simple and intuitive, yet valuable information to aid surgeons. Therefore, especially for young patients, holding out the lateral pillar through joint preserving procedures might be the key in preventing further collapse.

Improved Biological Responses of Titanium Coating Using Laser-Aided Direct Metal Fabrication on SUS316L Stainless Steel.

Direct metal fabrication (DMF) coatings have the advantage of a more uniform porous structure and superior mechanical properties compared to coatings provided by other methods. We applied pure titanium metal powders to SUS316L stainless steel using laser-aided DMF coating technology with 3D printing. The purpose of this study was to determine the efficacy of this surface modification of stainless steel. The capacity of cells to adhere to DMF-coated SUS316L stainless steel was compared with machined SUS316L stainless steel in vitro and in vivo. Morphological in vitro response to human osteoblast cell lines was evaluated using scanning electron microscopy. Separate specimens were inserted into the medulla of distal femurs of rabbits for in vivo study. The distal femurs were harvested after 3 months, and were then subjected to push-out test and histomorphometrical analyses. The DMF group exhibited a distinct surface chemical composition, showing higher peaks of titanium compared to the machined stainless steel. The surface of the DMF group had a more distinct porous structure, which showed more extensive coverage with lamellipodia from osteoblasts than the machined surface. In the in vivo test, the DMF group showed better results than the machined group in the push-out test (3.39 vs. 1.35 MPa, respectively, p = 0.001). In the histomorphometric analyses, the mean bone-to-implant contact percentage of the DMF group was about 1.5 times greater than that of the machined group (65.4 ± 7.1% vs. 41.9 ± 5.6%, respectively; p < 0.001). The porous titanium coating on SUS316L stainless steel produced using DMF with 3D printing showed better surface characteristics and biomechanical properties than the machined SUS316L.

Dysfunction of X-linked inhibitor of apoptosis protein (XIAP) triggers neuropathological processes via altered p53 activity in Huntington's disease.

Mitochondrial dysfunction is associated with neuronal damage in Huntington's disease (HD), but the precise mechanism of mitochondria-dependent pathogenesis is not understood yet. Herein, we found that colocalization of XIAP and p53 was prominent in the cytosolic compartments of normal subjects but reduced in HD patients and HD transgenic animal models. Overexpression of mutant Huntingtin (mHTT) reduced XIAP levels and elevated mitochondrial localization of p53 in striatal cells in vitro and in vivo. Interestingly, XIAP interacted directly with the C-terminal domain of p53 and decreased its stability via autophagy. Overexpression of XIAP prevented mitochondrially targeted-p53 (Mito-p53)-induced mitochondrial oxidative stress and striatal cell death, whereas, knockdown of XIAP exacerbated Mito-p53-induced neuronal damage in vitro. In vivo transduction of AAV-shRNA XIAP in the dorsal striatum induced rapid onset of disease and reduced the lifespan of HD transgenic (N171-82Q) mice compared to WT littermate mice. XIAP dysfunction led to ultrastructural changes of the mitochondrial cristae and nucleus morphology in striatal cells. Knockdown of XIAP exacerbated neuropathology and motor dysfunctions in N171-82Q mice. In contrast, XIAP overexpression improved neuropathology and motor behaviors in both AAV-mHTT-transduced mice and N171-82Q mice. Our data provides a molecular and pathological mechanism that deregulation of XIAP triggers mitochondria dysfunction and other neuropathological processes via the neurotoxic effect of p53 in HD. Together, the XIAP-p53 pathway is a novel pathological marker and can be a therapeutic target for improving the symptoms in HD.

AEG-1 Regulates TWIK-1 Expression as an RNA-Binding Protein in Astrocytes.

AEG-1, also called MTDH, has oncogenic potential in numerous cancers and is considered a multifunctional modulator because of its involvement in developmental processes and inflammatory and degenerative brain diseases. However, the role of AEG-1 in astrocytes remains unknown. This study aimed to investigate proteins directly regulated by AEG-1 by analyzing their RNA expression patterns in astrocytes transfected with scramble shRNA and AEG-1 shRNA. AEG-1 knockdown down-regulated TWIK-1 mRNA. Real-time quantitative PCR (qPCR) and immunocytochemistry assays confirmed that AEG-1 modulates TWIK-1 mRNA and protein expression. Electrophysiological experiments further revealed that AEG-1 further regulates TWIK-1-mediated potassium currents in normal astrocytes. An RNA immunoprecipitation assay to determine how AEG-1 regulates the expression of TWIK-1 revealed that AEG-1 binds directly to TWIK-1 mRNA. Furthermore, TWIK-1 mRNA stability was significantly increased upon overexpression of AEG-1 in cultured astrocytes (p < 0.01). Our findings show that AEG-1 serves as an RNA-binding protein to regulate TWIK-1 expression in normal astrocytes.

Long-Term Results of Total Hip Arthroplasty in Young Patients With Osteonecrosis After Allogeneic Bone Marrow Transplantation for Hematological Disease: A Multicenter, Propensity-Matched Cohort Study With a Mean 11-Year Follow-Up.

BACKGROUND: The number of young patients with hematological disease requiring total hip arthroplasty (THA) is expected to increase. We aimed to investigate the long-term THA outcomes in patients with osteonecrosis of the femoral head (ONFH) following allogeneic bone marrow transplantation (BMT) for hematological disease. METHODS: All patients who underwent THA for osteonecrosis after BMT from 1997 to 2012 were identified at 2 institutions. Using propensity scores, 75 THAs in 45 patients were matched for age, gender, body mass index, American Society of Anesthesiologists score, and year of surgery with 75 THAs in 58 patients with idiopathic ONFH without a history of hematological disease (1:1 ratio). The mean age at surgery was 36.7 years and 52% were men. Clinical and radiographic evaluations were performed and clinical scores were obtained at last follow-up. Kaplan-Meier analyses were used to compare survivorship. RESULTS: At a mean follow-up of 10.6 ± 3.5 years, clinical, radiographic, and survivorship outcomes, and the Harris hip scores were similar between both groups. The 13-year survivorship for all-cause revision was 93.4% for the BMT group and 95% for the control group (P = .928). No significant differences were observed between groups in the rates of reoperation (4% vs 5.3%, P = 1.000), 90-day readmission (all 5.3%), or overall mortality (4.4% vs 1.7%, P = .681). No hips had periprosthetic joint infection or septic loosening in either group. Osteolysis occurred in none of the BMT patients and in 2 hips (2.7%) of the control patients (P = .497). CONCLUSION: This large cohort multicenter survey at 11-year follow-up shows that contemporary cementless THA in young hematological disease patients after allogeneic BMT is not associated with a higher risk for surgical complications, revision, reoperation, readmission, and mortality compared to a matched cohort of idiopathic ONFH.

Comparative analysis of titanium coating on cobalt-chrome alloy in vitro and in vivo direct metal fabrication vs. plasma spraying.

BACKGROUND: Titanium surface coating on cobalt-chromium (CoCr) alloy has characteristics desirable for an orthopedic implant as follows: strength, osteointegrative capability, and biocompatibility. Creating such a coated surface takes a challenging process and two dissimilar metals are not easily welded. In our study, we utilized additive manufacturing with a 3D printing called direct metal fabrication (DMF) and compared it to the plasma spraying method (TPS), to coat titanium onto CoCr alloy. We hypothesized that this would yield a coated surface quality as acceptable or better than the already established method of plasma spraying. For this, we compared characteristics of titanium-coated surfaces created by direct metal fabrication method (DMF) and titanium plasma spraying (TPS), both in vitro and in vivo, for (1) cell morphology, (2) confocal microscopy images of immunofluorescent assay of RUNX2 and fibronectin, (3) quantification of cell proliferation rate, (4) push-out biomechanical test, and (5) bone histomorphometry. METHOD: For in vitro study, human osteoblast cells were seeded onto the coated surfaces. Cellular morphology was observed with a scanning electron microscope. Cellular proliferation was validated with ELISA, immunofluorescent assay. For in vivo study, coated rods were inserted into the distal femur of the rabbit and then harvested. The rods were biomechanically tested with a push-out test and observed for histomorphometry to evaluate the microscopic bone to implant ratio. RESULT: For cell morphology observation, lamellipodia and filopodia, a cytoplasmic projection extending into porous structure, formed on both surfaces created by DMF and TPS. The proliferation of the osteoblasts, the DMF group showed a better result at different optic density levels (p = 0.035, 0.005, 0.001). Expression and distribution of fibronectin and Runx-2 genes showed similar degrees of expressions. The biomechanical push-out test yielded a similar result (p = 0.714). Histomorphometry analysis also showed a similar result (p = 0.657). CONCLUSION: In conclusion, DMF is a method which can reliably create a proper titanium surface on CoCr alloy. The resulting product of the surface shows a similar quality to that of the plasma spraying method, both in vivo and in vitro, in terms of biological and mechanical property.

Synthesis and biological evaluation of quinoxaline derivatives as specific c-Met kinase inhibitors.

A series of novel quinoxaline derivatives were synthesized and evaluated for their inhibitory activity against c-Met kinase enzyme. Most of the tested compounds exhibited potent inhibitory activity. All the synthesized quinoxaline compounds were further examined against c-Met overexpressed human gastric cancer cell line (MKN-45), which showed good inhibitory activity. Among the synthesized compounds, compound 4 exhibited better tumor growth inhibition in the animal model study; we also confirmed its acceptable drug property and highly selective target activity.

The Knockdown of TREK-1 in Hippocampal Neurons Attenuate Lipopolysaccharide-Induced Depressive-Like Behavior in Mice.

TWIK-related potassium channel-1 (TREK-1) is broadly expressed in the brain and involved in diverse brain diseases, such as seizures, ischemia, and depression. However, the cell type-specific roles of TREK-1 in the brain are largely unknown. Here, we generated a Cre-dependent TREK-1 knockdown (Cd-TREK-1 KD) transgenic mouse containing a gene cassette for Cre-dependent TREK-1 short hairpin ribonucleic acid to regulate the cell type-specific TREK-1 expression. We confirmed the knockdown of TREK-1 by injecting adeno-associated virus (AAV) expressing Cre into the hippocampus of the mice. To study the role of hippocampal neuronal TREK-1 in a lipopolysaccharide (LPS)-induced depression model, we injected AAV-hSyn-BFP (nCTL group) or AAV-hSyn-BFP-Cre (nCre group) virus into the hippocampus of Cd-TREK-1 KD mice. Interestingly, the immobility in the tail suspension test after LPS treatment did not change in the nCre group. Additionally, some neurotrophic factors (BDNF, VEGF, and IGF-1) significantly increased more in the nCre group compared to the nCTL group after LPS treatment, but there was no difference in the expression of their receptors. Therefore, our data suggest that TREK-1 in the hippocampal neurons has antidepressant effects, and that Cd-TREK-1 KD mice are a valuable tool to reveal the cell type-specific roles of TREK-1 in the brain.

Titanium Powder Coating Using Metal 3D Printing: A Novel Coating Technology for Cobalt-Chromium Alloy Implants.

Background: Three-dimensional (3D) printing with a direct metal fabrication (DMF) technology has been innovatively introduced in the field of surface treatment of prostheses. The purpose of this study was to determine whether such modifications on the surface of cobalt-chromium (CoCr) alloy by titanium powder coating using DMF improves the osseointegration ability of CoCr alloy. Methods: We compared the in vitro and in vivo ability of cells to adhere to DMF-coated CoCr alloy with machining. Biological and morphological responses to human osteoblast cell lines were examined by measuring cell proliferation rate and observing expression of actin filament. For in vivo study, we inserted different specimens in each medulla of the distal femurs of rabbit. After 3 months, the distal femurs were harvested, and a push-out test and histomorphometric analyses were performed. Results: The cell proliferation rate and cell adhesion in the DMF group were higher compared with those in the machined group. Human osteoblast cells on the DMF-coated surface were more strongly adhered and well-proliferated compared with those on the other surface. In the in vivo test, there was a significant difference in the ultimate shear strength between the DMF and machined groups (2.49 MPa vs. 0.87 MPa, respectively, p = 0.001). In the histomorphometric analysis, there was a significant difference in the mean bone-to-implant contact percentages between the DMF and machined groups (72.3 ± 6.2% vs. 47.6 ± 6.9%, respectively, p < 0.001). Conclusion: Titanium coating of CoCr alloy with 3D metal printing provides optimal surface characteristics and a good biological surface both in vitro and in vivo.

Surgical accuracy, function, and quality of life of simultaneous versus staged bilateral Total hip Arthroplasty in patients with Osteonecrosis of the femoral head.

BACKGROUND: The optimal surgical option for patients requiring bilateral hip replacement remains controversial. The purpose of this study was to compare surgical accuracy; functional outcome and health-related quality of life; and prosthetic-related complications and revision surgery of a simultaneous bilateral total hip arthroplasty (THA) with those of a staged bilateral THA with an interval between procedures <12 months. METHODS: A total of 123 unselected consecutive patients (mean age, 43.3 years) who underwent bilateral THAs for osteonecrosis of the femoral head (ONFH) with a minimum follow-up of two years (mean, 60.2 months) were studied retrospectively; 63 simultaneous procedures served as a test group and 60 staged procedures served as a control group. RESULTS: The mean postoperative leg-length discrepancy (LLD) and the percentage of patients who had an LLD >3 mm were significantly lower in the simultaneous group (P < 0.001 and P = 0.001, respectively). A higher number of cups within the safe zones, a higher correction rate, and a lower failure rate for the cup placement in the second-operated hip were also identified in the simultaneous group. The mean Harris hip score, EuroQol-5D index, and EuroQol-visual analogue scale score were all better in the simultaneous group at the latest follow-up (P < 0.001, in all comparisons). We found that the simultaneous procedure was associated with a lower incidence of postoperative prosthetic-related complications and revision surgery. CONCLUSIONS: We suggest that bilateral ONFH could be treated with a simultaneous THA rather than a staged THA to achieve a better surgical outcome.