TGF-ß Type I Receptor Kinase Inhibitor EW-7197 Suppresses Cholestatic Liver Fibrosis by Inhibiting HIF1α-Induced Epithelial Mesenchymal Transition.

BACKGROUND/AIMS: Hypoxia is an environmental factor that aggravates liver fibrosis. HIF1α activates hepatic stellate cells (HSCs) and increases transforming growth factor-ß (TGF-ß) signaling and the epithelial mesenchymal transition (EMT), accelerating the progression of fibrosis. We evaluated the anti-fibrotic therapeutic potential of a small-molecule inhibitor of TGF-ß type I receptor kinase, EW-7197, on HIF1α-derived TGF-ß signaling in cholestatic liver fibrosis. METHODS: We used a bile duct ligation (BDL)-operated rat model to characterize the role of HIF1α-derived TGF-ß signaling in liver fibrosis. Cellular assays were performed in LX-2 cells (human immortalized HSCs). The anti-fibrotic effects of EW-7197 in liver tissues and HSCs were investigated via biochemical assays, immunohistochemistry (IHC), immunofluorescence (IF), chromatin immunoprecipitation (ChIP) assays, real-time PCR, and western blotting. RESULTS: In our BDL rat model, orally administered EW-7197 inhibited fibrosis and attenuated HIF1α-induced activation of HSCs and EMT in vivo. In addition, EW-7197 inhibited HIF1α-derived HSC activation and expression of EMT markers in LX-2 cells in vitro. CONCLUSION: This study suggests that EW-7197 exhibits potential as a treatment for liver fibrosis because it inhibits HIF1α-induced TGF-ß signaling.

EW-7197 inhibits hepatic, renal, and pulmonary fibrosis by blocking TGF-ß/Smad and ROS signaling.

Fibrosis is an inherent response to chronic damage upon immense apoptosis or necrosis. Transforming growth factor-beta1 (TGF-ß1) signaling plays a key role in the fibrotic response to chronic liver injury. To develop anti-fibrotic therapeutics, we synthesized a novel small-molecule inhibitor of the TGF-ß type I receptor kinase (ALK5), EW-7197, and evaluated its therapeutic potential in carbon tetrachloride (CCl4) mouse, bile duct ligation (BDL) rat, bleomycin (BLM) mouse, and unilateral ureteral obstruction (UUO) mouse models. Western blot, immunofluorescence, siRNA, and ChIP analysis were carried out to characterize EW-7197 as a TGF-ß/Smad signaling inhibitor in LX-2, Hepa1c1c7, NRK52E, and MRC5 cells. In vivo anti-fibrotic activities of EW-7197 were examined by microarray, immunohistochemistry, western blotting, and a survival study in the animal models. EW-7197 decreased the expression of collagen, α-smooth muscle actin (α-SMA), fibronectin, 4-hydroxy-2, 3-nonenal, and integrins in the livers of CCl4 mice and BDL rats, in the lungs of BLM mice, and in the kidneys of UUO mice. Furthermore, EW-7197 extended the lifespan of CCl4 mice, BDL rats, and BLM mice. EW-7197 blocked the TGF-ß1-stimulated production of reactive oxygen species (ROS), collagen, and α-SMA in LX-2 cells and hepatic stellate cells (HSCs) isolated from mice. Moreover, EW-7197 attenuated TGF-ß- and ROS-induced HSCs activation to myofibroblasts as well as extracellular matrix accumulation. The mechanism of EW-7197 appeared to be blockade of both TGF-ß1/Smad2/3 and ROS signaling to exert an anti-fibrotic activity. This study shows that EW-7197 has a strong potential as an anti-fibrosis therapeutic agent via inhibition of TGF-ß-/Smad2/3 and ROS signaling.

Radiological and clinical outcomes after Attune primary total knee arthroplasty using Stemmable Tibia: A two-year follow-up prospective bi-center study.

This prospective bi-center study aimed to analyze the outcomes of primary total knee arthroplasty using the Stemmable Tibia Attune system. A total of 100 patients who underwent primary total knee arthroplasty with Stemmable Tibia from January 2019 to December 2021 were enrolled in the study. Radiological outcomes (hip-knee-ankle axis and medial proximal tibial angle) were assessed preoperatively and postoperatively. Clinical outcomes (visual analog scale score, Hospital for Special Surgery score, Knee Society function score, Knee Society knee score, flexion contracture, further flexion, and range of motion) were analyzed preoperatively and at 6 weeks, 3 months, 6 months, 1 year, and 2 years postoperatively. Complications (periprosthetic joint infection and aseptic loosening) were examined. The hip-knee-ankle axis decreased (preoperative: 9.5° ± 6.3°, postoperative: 1.1° ± 2.7°), whereas the medial proximal tibial angle increased (preoperative: 84.6° ± 4.1°, postoperative: 89.8° ± 1.9°). The visual analog scale score, Hospital for Special Surgery score, Knee Society knee score, and Knee Society function score increased postoperatively. The Knee Society knee score indicated above good outcomes (100.0% and 99.0% at 1 and 2 years postoperatively, respectively). The Knee Society function score also showed above good results (98.0% and 93.0% at 1 and 2 years postoperatively, respectively). The range of motion significantly improved (p < 0.001): flexion contracture decreased from 9.10° ± 7.23° to 2.15° ± 2.87°, whereas further flexion increased from 136.05° ± 14.78° to 139.80° ± 10.02°. One patient developed periprosthetic joint infection; no early loosening was observed. In conclusion, Attune primary total knee arthroplasty with Stemmable Tibia not only is safe and effective but also leads to radiological and clinical improvements.

Recovery After Hip Arthroscopy in Patients With Combined Femoroacetabular Impingement and Labral Tears Compared With Isolated Pathology.

Background: Outcomes for patients with femoroacetabular impingement (FAI) treated with hip arthroscopy can differ depending on whether there is underlying intra-articular pathology. Purpose: To evaluate the outcomes of patients after undergoing hip arthroscopy depending on their underlying pathology (isolated FAI, isolated labral tear, or combined FAI and a labral tear) using the 12-Item International Hip Outcome Tool (iHOT-12). Study Design: Cohort study; Level of evidence, 3. Methods: A total of 75 patients diagnosed with FAI with or without labral tears and isolated labral tears who underwent hip arthroscopy performed by the same surgeon at a single institution from January 2014 to December 2019 were included in this study. All patients had at least 2 years of follow-up data. Patients were divided into 3 groups as follows: patients with FAI and an intact labrum; patients with an isolated labral tear; and patients with combined FAI and a labral tear. The iHOT-12 scores at 1.5, 3, 6, 12, 18, and >24 months postoperatively were compared and analyzed. Outcome scores were also evaluated in terms of the substantial clinical benefit (SCB) and the patient-acceptable symptomatic state (PASS). Results: Of 75 patients who underwent hip arthroscopy, 14 had FAI, 23 had labral tears, and 38 had both. All groups showed significant improvements on the iHOT-12 from preoperative to the final follow-up (FAI, from 37.64 ± 3.77 to 93.64 ± 1.50; labral tear, from 33.70 ± 3.55 to 93 ± 1.24; combined, from 28.55 ± 3.15 to 93.03 ± 0.88) (P < .001 for all). However, compared with other groups, the patients with FAI and a labral tear had lower scores at 1.5, 3, 6, and 12 months postoperatively (P < .001), highlighting a slower rate of recovery. For all groups, recovery to normal function according to the SCB was 100% at 12 months, and satisfaction according to the PASS was 100% at 18 months postoperatively. Conclusion: The final iHOT-12 scores were similar at 18 months regardless of the pathology treated; however, patients with FAI and a labral tear took longer to reach their plateau.

Safety of using a large femoral head on thin polyethylene for total hip arthroplasty based on different types of polyethylene.

The use of a large femoral head in total hip arthroplasty (THA) to stabilize and reduce the incidence of dislocation is on the increase, but concerns arise when combining them with small acetabular components due to potential mechanical failures in thin polyethylene (PE) liners. A single-institution, retrospective cohort study was conducted on 116 patients with minimum 2-year follow-up who received 36-mm femoral heads and acetabular components ≤ 52 mm, using either remelted highly cross-linked polyethylene (remelted HXLPE) or vitamin E-infused HXLPE (VEPE). Osteolysis and implant loosening were not observed in either group. Although a fracture of the PE liner was observed in each group (1.7%), the clinical outcomes were excellent, as the mean modified Harris Hip Score (mHHS) at the last follow-up was 93.5. Moreover, the mean linear wear rates measured by digital imaging software in both groups were low, with 0.035 mm/y in remelted HXLPE and 0.030 mm/y in VEPE. In conclusion, The use of a large femoral head on a thin PE liner can be a viable treatment option in patients who need to prioritize stability; however, careful attention should be paid to mechanical fractures of the PE liner.

The use of dual mobility acetabular cups in total hip replacement reduces dislocation rates in hip dysplasia patients.

Total hip replacement arthroplasty (THA) in hip dysplasia patients has a higher dislocation rate than in patients with simple hip osteoarthritis due to anatomical deformation. Therefore, to reduce postoperative THA dislocation is the challenge for arthroplasty surgeons. From 2015 to 2020, 1525 patients underwent THA performed by two surgeons at a single institution. A total of 152 patients involving 172 THAs were included. The patients were classified into dual-mobility (DM) and fixed-bearing (FB) acetabular cup groups. The occurrence of postoperative dislocation and functional evaluation of the hip joint, was analyzed before and after surgery using the modified Harris hip score(mHHS). There was no difference in the preoperative demographics and radiographic parameters between the groups. The incidence of postoperative hip dislocation was significantly lower in the DM group (DM 0% vs. FB 9.0%) (P value = 0.003). The mHHS showed no difference before surgery and after surgery (DM 91.80 vs FB 92.03). Treating hip dysplasia patients with THA using a dual-mobility acetabular cup can reduce postoperative dislocations, and could be used for the better management of these patients.

The usefulness of dual mobility cups in primary total hip arthroplasty patients at a risk of dislocation.

This study aimed to evaluate the early results of primary total hip arthroplasty (THA) using dual mobility (DM) cups in patients at a risk of dislocation and compare them with that of fixed bearing (FB) THA. This retrospective study included patients who had undergone primary THA between January 2016 and December 2018 and were at a risk of dislocation. A propensity score-matched analysis was conducted for 63 THA procedures with vitamin-E infused highly cross-linked polyethylene (VEPE) DM bearing and 63 THA procedures performed with FB from the same manufacturer for a mean follow-up period of 3.1 and 3.5 years, respectively. The radiologic outcomes at the last follow-up and incidence of postoperative complications were evaluated and compared statistically between the two groups. The modified Harris hip score (mHHS) was used to assess patient-reported outcomes. Postoperative dislocation occurred in 4 cases (6.3%) in the FB group, but did not occur in the DM group (p = 0.042). There was no difference in the radiologic outcomes and postoperative complications between the two groups. The mHHS at the last follow-up showed satisfactory outcomes in both the groups (DM group, 90.5; FB group, 88.1), without a statistical difference between the groups. The early results of THA using VEPE DM bearing showed better outcomes than that of THA with FB for patients at a risk of dislocation. A longer follow-up period is recommended to assess the stability and overall outcomes.

Cementless total hip arthroplasty following failed internal fixation for femoral neck and intertrochanteric fractures: A comparative study with 3-13 years' follow-up of 96 consecutive patients.

BACKGROUND: The aim of this study was to investigate the postoperative outcomes of cementless Total hip arthroplasty (THA) following failed internal fixation for femoral neck and intertrochanteric fractures. METHOD: Ninety-six cementless THAs for failed internal fixation after femoral neck fracture (59, group I) and intertrochanteric fracture (37, group II) with a minimum follow-up of 3 years were analyzed. Clinical and radiologic evaluations were performed on all patients. RESULTS: The intraoperative blood loss and operating time were significantly increased in group II (p = 0.001, p = 0.001, respectively). Harris hip score at last follow-up was significantly improved in group I (p = 0.007) but, there were no differences in hospital stay, Koval score at last follow-up, and perioperative complications between both groups. Long femoral stems for diaphyseal fitting were frequently used in group II (32/37, 86%) (p = 0.001). Radiographically, none of the acetabular cups showed evidence of migration, loosening. All cases showed stable fixation of the femoral stem at last follow-up. CONCLUSIONS: Outcomes of cementless THA following failed internal fixation for femoral neck and intertrochanteric fractures were satisfactory; increased intraoperative blood loss, operating time, and requirement of long femoral stem should be considered in the latter type of fracture.

TIMP-1 mediates TGF-ß-dependent crosstalk between hepatic stellate and cancer cells via FAK signaling.

Transforming growth factor-ß (TGF-ß) signaling plays a key role in progression and metastasis of HCC. This study was undertaken to gain the proof of concept of a small-molecule inhibitor of TGF-ß type I receptor kinase, EW-7197 as a potent anti-cancer therapy for HCC. We identified tissue inhibitors of metalloproteinases-1 (TIMP-1) as one of the secreted proteins of hepatic stellate cells (HSCs) and a key mediator of TGF-ß-mediated crosstalk between HSCs and HCC cells. TGF-ß signaling led to increased expression of TIMP-1, which activates focal adhesion kinase (FAK) signaling via its interaction with CD63. Inhibition of TGF-ß signaling using EW-7197 significantly attenuated the progression and intrahepatic metastasis of HCC in an SK-HEP1-Luc orthotopic-xenograft mouse model. In addition, EW-7197 inhibited TGF-ß-stimulated TIMP-1 secretion by HSCs as well as the TIMP-1-induced proliferation, motility, and survival of HCC cells. Further, EW-7197 interrupted TGF-ß-mediated epithelial-to-mesenchymal transition and Akt signaling, leading to significant reductions in the motility and anchorage-independent growth of HCC cells. In conclusion, we found that TIMP-1 mediates TGF-ß-regulated crosstalk between HSCs and HCC cells via FAK signaling. In addition, EW-7197 demonstrates potent in vivo anti-cancer therapeutic activity and may be a potential new anti-cancer drug of choice to treat patients with liver cancer.

Combinatorial TGF-ß attenuation with paclitaxel inhibits the epithelial-to-mesenchymal transition and breast cancer stem-like cells.

Distant relapse after chemotherapy is an important clinical issue for treating breast cancer patients and results from the development of cancer stem-like cells (CSCs) during chemotherapy. Here we report that blocking epithelial-to-mesenchymal transition (EMT) suppresses paclitaxel-induced CSCs properties by using a MDA-MB-231-xenografted mice model (in vivo), and breast cancer cell lines (in vitro). Paclitaxel, one of the cytotoxic taxane-drugs such as docetaxel, increases mesenchymal markers (Vimentin and Fibronectin) and decreases an epithelial marker (Zo-1). Blocking TGF-ß signaling with the TGF-ß type I receptor kinase (ALK5) inhibitor, EW-7197, suppresses paclitaxel-induced EMT and CSC properties such as mammosphere-forming efficiency (MSFE), aldehyde dehydrogenase (ALDH) activity, CD44+/CD24- ratio, and pluripotency regulators (Oct4, Nanog, Klf4, Myc, and Sox2). The combinatorial treatment of EW-7197 improves the therapeutic effect of paclitaxel by decreasing the lung metastasis and increasing the survival time in vivo. We confirmed that Snail is increased by paclitaxel-induced intracellular reactive oxygen species (ROS) and EW-7197 suppresses the paclitaxel-induced Snail and EMT by attenuating paclitaxel-induced intracellular ROS. Knock-down of SNAI1 suppresses paclitaxel-induced EMT and CSC properties. These data together suggest that blocking the Snail-induced EMT with the ALK5 inhibitor attenuates metastasis after paclitaxel-therapy and that this combinatorial approach could prove useful in treating breast cancer.

Daily grape juice consumption reduces oxidative DNA damage and plasma free radical levels in healthy Koreans.

Grape contains flavonoids with antioxidant properties which are believed to be protective against various types of cancer. This antioxidative protection is possibly provided by the effective scavenging of reactive oxygen species (ROS), thus defending cellular DNA from oxidative damage and potential mutations. This study of healthy adults tested whether a daily regimen of grape juice supplementation could reduce cellular DNA damage in peripheral lymphocytes and reduce the amount of free radicals released. Sixty-seven healthy volunteers (16 women and 51 men) aged 19-57 years were given 480 ml of grape juice daily for 8 weeks in addition to their normal diet, and blood samples were drawn before and after the intervention. The DNA damage was determined by using the single cell gel (comet) assay with alkaline electrophoresis and was quantified by measuring tail length (TL). Levels of free radicals were determined by reading the lucigenin-perborate ROS generating source, using the Ultra-Weak Chemiluminescence Analyzer System. Grape juice consumption resulted in a significant decrease in lymphocyte DNA damage expressed by TL (before supplementation: 88.75 +/- 1.55 microm versus after supplementation: 70.25 +/- 1.31 microm; P=0.000 by paired t-test). Additionally, grape juice consumption for 8 weeks reduced the ROS/photon count by 15%, compared to the beginning of the study. The preventive effect of grape juice against DNA damage was simultaneously shown in both sexes. These results indicate that the consumption of grape juice may increase plasma antioxidant capacity, resulting in reduced DNA damage in peripheral lymphocytes achieved at least partially by a reduced release of ROS. Our findings support the hypothesis that polyphenolic compounds contained in grape juice exert cancer-protective effects on lymphocytes, limiting oxidative DNA damage possibly via a decrease in free radical levels.

Concord grape juice supplementation reduces blood pressure in Korean hypertensive men: double-blind, placebo controlled intervention trial.

Many of the flavonoids found in grapes and grape products such as juice or wine have been known to exert antioxidant, anti-inflammatory, platelet inhibitory and arterial relaxing effects either in vitro, in animal studies and in human trials. This study was designed to test the effect of Concord grape juice consumption on altering blood pressure in hypertensive patients. Forty subjects were given 5.5 ml/kg body weight/day of either Concord grape juice (CGJ) or a calorie-matched placebo drink every day for 8 weeks. Blood pressure (BP) was measured on weeks 0, 4 and 8. Compared to baseline, in the CGJ group systolic BP was reduced on average by 7.2 mm Hg (p = 0.005) and diastolic BP was reduced on average by 6.2 mm Hg (p = 0.001) at the end of 8 weeks. Comparable changes in the group getting the placebo product were -3.5 mm Hg (NS) and -3.2 mm Hg (p = 0.05) Consuming Concord grape juice, which is high in polyphenolic compounds, may favorably affect BP in hypertensive individuals.

Changes in antioxidant status, blood pressure, and lymphocyte DNA damage from grape juice supplementation.

In a placebo-controlled double-blinded study, the effect of 8 weeks of grape juice was compared to the effect of isocaloric placebo juice. Volunteers with a systolic blood pressure > 130 mmHg and diastolic blood pressure > 90 mmHg were recruited. A total of 40 healthy subjects were randomized to receive isocaloric juices for 8 weeks. Twenty-one subjects were instructed to consume 5.5 mL/kg daily of grape juice (GJ), and 19 subjects consumed placebo juice (PJ). Plasma antioxidant vitamin C, total radical trapping antioxidant capacity, blood pressure, and lymphocyte DNA damage were assessed pre- and postsupplementation. Plasma total radical-trapping antioxidant potential showed an increase at the level of 1.31 +/- 0.01 (postsupplementation) versus 1.33 +/- 0.01 (presupplementation) (P < 0.1). Grape juice consumption resulted in a 26% decrease in lymphocyte DNA (both hydrogen peroxide treated or spontaneous) in the grape juice group, while no difference was found in the PJ group. Consuming moderate amounts of daily grape juice may favorably affect antioxidant defense systems and lymphocyte DNA damage in hypertensive individuals.