Salubrinal protects against inflammatory response in macrophage and attenuates psoriasiform skin inflammation by antagonizing NF-κB signaling pathway.

Psoriasiform skin inflammation is the common chronic skin inflammatory disease with no effective clinical therapy. Salubrinal is a multifunctional molecule playing a protective role in several conditions. Recently, studies have reported that Salubrinal is a potential therapeutic agent for inflammatory diseases. However, the protective role of Salubrinal in psoriasis-like skin inflammation remains unknown. In this article, imiquimod (IMQ)-induced psoriasis models were established in wild-type mice to explore the role of Salubrinal in the development of psoriasis. As a result, the IMQ-induced mouse models exhibited typical skin inflammation, which was alleviated by the administration of Salubrinal. Furthermore, RAW264.7 macrophage was stimulated with Lipopolysaccharide(LPS) in the presence or absence of Salubrinal. LPS stimulation elevated the expression of various inflammatory biomarkers, while the administration of Salubrinal abolished the function of LPS in RAW264.7 macrophages. In addition, the activation of the nuclear factor-kappa B (NF-κB) signaling pathway in both the LPS-stimulated RAW264.7 macrophage and psoriasis mouse models was antagonized by the administration of Salubrinal. Collectively, Salubrinal might be considered as a promising therapeutic agent for psoriasis-like skin inflammation.

Formulation and characterization of lornoxicam-loaded cellulosic-microsponge gel for possible applications in arthritis.

Rheumatoid arthritis (RA) is an autoimmune disease associated with severe joint pain. Herein, we report lornoxicam loaded cellulosic microsponge gel formulation with sustained anti-inflammatory effects that are required to manage arthritic pain. The microsponges were formulated using quasi emulsion-solvent diffusion method employing four different surfactant systems, namely polyvinyl alcohol (PVA), Tween80, Gelucire 48/16 and Gelucire 50/13. All the lornoxicam loaded microsponge formulations were extensively characterized with a variety of analytical tools. The optimized microsponge formulation was then converted into gel formulation. The lornoxicam loaded microsponge gel formulation had adequate viscosity and sufficient pharmaceutical properties as confirmed by the texture analysis and the drug release followed Super case II transport. It is noteworthy that we described the preparation of a new cellulosic polymers based microsponge system for delivery of lornoxicam to provide quick as well as lasting (sustained) anti-inflammatory effects in rats using carrageenan induced rat paw edema model. We were able to demonstrate a 72% reduction in inflammation within 4 h using the optimize transdermal gel formulation utilizing Transcutol P as permeation enhancer and with the aid of skin micro-piercing by microneedles, hence, demonstrating the potential of this microsponge gel formulation in arthritis management.

Engineered High-Loaded Mixed-Monoclonal Antibodies (Adalimumab, Rituximab and Trastuzumab) Polymeric Nanoparticle for Rheumatoid Arthritis Treatment: A Proof of Concept.

Rheumatoid arthritis, a chronic disease, affects from 0.5% to 1% of the world population. The main consequences include loss of joint functionality and severe pain, with lost in life quality and increased risk of morbidity and mortality. The main strategy for RA treatment relies in early diagnosis as targeted therapy. In this regard, the development and application of designed/engineered nanoparticles may represent an innovative approach and the key to success, since is a personalized nanodrug. Thus, we have synthetized, characterized, and in vivo evaluated a tri-loaded monoclonal antibody nanoparticle. For the production we used a mix of monoclonal antibodies: adalimumab, rituximab and trastuzumab to surround all RA metabolic pathways. The characterization included atomic force microscopy, dynamic light scattering analysis and entrapment efficacy using BCA analysis. The in vivo evaluation was done in mice. At this stage we used animals to assess the pharmacokinetics, the tissue distribution as the proof of concept (therapeutic efficacy) of the nanoparticles developed in inducted animals with rheumatoid arthritis. The interpretation of our results revealed that a spherical shaped nanoparticle has been produced with a mean size of 229.7 nm, and a polydispersity index of 0.191. This data has been corroborated by DLS and AFM analysis. The pre-clinical (in vivo) evaluation demonstrated a low elimination rate of 2,34 L/hour, with a purge of 0,42 h. The therapeutic efficacy showed that the nanoparticles have an increased therapeutic effect than the conventional drug with a reduction in all main parameters including the interleukins.

Modified arthroscopic remplissage for Hill-Sachs lesions with high-strength sutures.

OBJECTIVE: The aim is to present a modified arthroscopic remplissage for shoulder Hill-Sachs lesions with high-strength sutures instead of suture anchors, to achieve better tendon-bone healing and avoid failure of remplissage due to anchor detachment. MATERIAL AND METHODS: A total of seven patients with recurrent anterior shoulder dislocation combined with a Hill-Sachs lesion were included in this study. Firstly, anteroinferior glenoid labrum complex damage was treated then 2-3 bone tunnels were punched with a sighting device from the bony defect of the humeral head to the inside of lesser tubercles of the humerus. The bony defect was filled by stitching the infraspinatus tendon through the bony tunnels with high-strength sutures. After the operation, the filling and healing of the infraspinatus tendon in the Hill-Sachs lesion were assessed using magnetic resonance imaging (MRI). RESULTS: Patients were followed up for 12 months. The results of MRI showed that all of the filled tendons healed well. Postoperative external rotation of the shoulder joint increased on average from 67° to 87°. Compared with the preoperative level, the Oxford Shoulder Instability Score (OSIS) was 18.50 ± 1.04 points higher and the Rowe score was increased by 66.755 ± 0.914 points. CONCLUSION: Arthroscopic remplissage of a shoulder Hill-Sachs lesion with high-strength sutures carries the benefits of secure fixing and good tendon-bone healing without the risk of anchor detachment.

Combined anteversion technique in total hip arthroplasty for Crowe IV developmental dysplasia of the hip.

BACKGROUND: A high rate of postoperative dislocation in total hip arthroplasty (THA) for Crowe IV developmental dysplasia of the hip (DDH) has been reported, 1 of the main reasons being higher true acetabular anteversion. If the cup is fixed with normal anteversion, the anterior rim will be excessively exposed, which reduces the contact areas of the cup and bone, affects prosthesis stability, and leads to iliopsoas tendinitis and persistent hip pain after THA. The aim of this study was to demonstrate that when cup anteversion is larger, adjusting femoral anteversion to bring the combined anteversion (CA) into the "safe zone" might prevent dislocation. METHODS: After having fixed the cup in the acetabulum according to the patients' native acetabular anteversion, we shortened and rotated the proximal femur to reduce femoral anteversion, adjusting the CA into the "safe zone". The Harris Hip Score (HHS) was used to evaluate hip joint function. Computerised tomography scanning was used to measure the anteversion angles. RESULTS: All patients were followed up without any dislocation. Preoperative and 12 months after surgery, the mean HHS were 43.3 ± 2.6 (38-47) and 88.1 ± 3.3 (78-92) respectively. Pre- and post-operation, the mean CA angles were 88.6° ± 9.4° (80.3°-119.4°) and 49.2° ± 2.6° (43.4°-54.4°) respectively. The bone healing time of femoral osteotomy ranged from 4 to 14 months, with a mean time of 7.5 months. CONCLUSIONS: This CA technique in THA for Crowe IV DDH can effectively prevent postoperative dislocation and provide good hip function.

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma.

Immunotherapy with tumor lysate-pulsed dendritic cells (DCs) is one of the breakthrough strategies used in the treatment of cancer. However, DC-based immunotherapies for osteosarcoma are limited. In the present study, preclinical studies of a C3H osteosarcoma mouse model (produced by subcutaneous injection of LM8 murine osteosarcoma cells) validated the concept that LM8 cell lysate-pulsed bone marrow-derived DCs may evoke a more potent immune response compared with DCs that have been matured using polyinosinic:polycytidylic acid (poly I:C). A cytotoxic T lymphocyte (CTL) response was established using two groups of C3H mice (n=9) with osteosarcoma; the treatment group consisted of LM8 cell lysate-pulsed DCs and the control group consisted of DCs matured using poly I:C. Each group was immunized with doses of 1×106 cells twice per week for 3 weeks. No difference in the expression of cluster of differentiation markers was identified in the two groups. DCs pulsed with LM8 cell lysate were associated with the increased induction of CTL activity. Serum interferon-γ levels were increased in mice that received DCs pulsed with LM8 cell lysate compared with that in the poly I:C-matured DC group (P<0.041). Serum interleukin-4 was decreased in the treatment group vs. the control group (P<0.033). A mixed lymphocyte reaction assay confirmed that LM8-DC immunotherapy may evoke a significant antigen-specific immune response in a mouse model. The present study reveals promising data on efficacy of a DC-based immunotherapy in the treatment of osteosarcoma; however, further clinical studies are warranted.

Exposing the transverse acetabular ligament in the presence of large floor osteophytes in primary total hip arthroplasty.

BACKGROUND/OBJECTIVE: The transverse acetabular ligament (TAL) can be used to position the acetabular cup and may help to improve the accuracy of primary total hip arthroplasty (THA). However, because the TAL may be covered by osteophytes, the ability to find the TAL varies greatly in the reported literature. In the present study, we introduce 2 methods and make a comparison between them to identify the easier procedure for finding an osteophyte-covered TAL. METHODS: During primary THA operations conducted from January 2012 to June 2015, a total of 100 patients (100 hips) were confirmed to have an osteophyte-covered TAL following the exposure of the acetabulum and removal of all soft tissues covering the TAL. These 100 patients were enrolled in this study. 2 methods were used to identify the TAL: the use of a bone chisel or a small reamer, and patients were allocated randomly to 1 of these 2 methods. The proportion of patients in whom TAL was identified was compared between the 2 methods using the chi-square test. RESULTS: The percentage of patients in whom the TAL was found using a bone chisel was 54.1% (26/48), whereas the percentage was 94.2% (49/52) in patients for whom a small reamer was used. The difference between the 2 methods was statistically significant (chi-square test, p<0.05). CONCLUSIONS: The TAL can be found more easily with a small reamer than with a bone chisel.

Effects of intraarticular (32)P colloid in the treatment of hemophilic synovitis of the knee: A short term clinical study.

BACKGROUND: Chronic synovitis is a consequence of recurrent intraarticular hemorrhage in patients with hemophilia. Eventually, synovitis leads to degeneration of the articular cartilage, with serious consequences that impact the quality-of-life in hemophiliacs. The aim of our study was to investigate the short term clinical effects of intraarticular injection of the radionuclide preparation(32)P colloid ((32)P-labelled colloidal chromic phosphate suspension) on recurrent intraarticular hemorrhages in patients with hemophilic synovitis of the knee. MATERIALS AND METHODS: Patients who met the inclusion criteria (n = 22) were enrolled in an open-label study between October 2011 and September 2012.(32)P colloid was injected into the knee joint and patients were followed up over 6 months after treatment. Hemorrhage frequency, visual analog scale pain score, hospital for special surgery knee score, knee circumference, upper knee circumference, knee diameter, and knee range of motion (ROM) were compared before and after treatment with intraarticular(32)P colloid injection. RESULTS: In 24 knees evaluated in 22 participating patients, there was a significant reduction in the number of hemorrhages after(32)P colloid treatment, along with significant pain relief. However, there were no statistically significant changes in the degree of joint swelling, degree of muscle atrophy and knee ROM between the pre and post treatment evaluations. CONCLUSION: The frequency of joint hemorrhage in patients with hemophilic knee synovitis can be significantly reduced and local symptoms can be improved in the short term by intraarticular injection of(32)P colloid.

Expression of TIM-3 on CD4+ and CD8+ T cells in the peripheral blood and synovial fluid of rheumatoid arthritis.

Rheumatoid arthritis (RA) is characterized by a chronic inflammatory process that targets the synovial lining of diarthrodial joints. TIM-3 plays a key role in the negative regulation of the immune response. In this study, we investigated the expression of TIM-3 on CD4+ and CD8+ T cells from systemic (peripheral blood) and local (synovial fluid) perspectives of RA. Level of TIM-3+ cells from peripheral blood and synovial fluid of patients as well as peripheral blood of healthy controls was measured by flow cytometry. Results showed that TIM-3 expression was significantly increased in both CD4+ and CD8+ T cells in the peripheral blood of RA (p < 0.001 and p < 0.001, respectively). Furthermore, patients revealed even higher expression of TIM-3 in CD4+ and CD8+ T cells in synovial fluid than in peripheral blood. When comparing TIM-3 level with the severity of RA, we identified that the percentage of TIM-3 on both peripheral CD4+ and peripheral CD8+ T cells was negatively correlated with disease activity score 28 (DAS28) of the patients. Similarly, TIM-3 on synovial fluid CD4+ and CD8+ T cells also revealed inverse correlation with DAS28 of the cases. Our data demonstrate a negative correlation between TIM-3 and the disease progression of RA.

Ginkgo biloba attenuates oxidative DNA damage of human umbilical vein endothelial cells induced by intermittent high glucose.

Intermittent high glucose (IHG), one of the general and important symptoms of patients with diabetes, has greater effect than sustained high glucose on the development of diabetic cardiovascular complications, in which endothelial dysfunction caused by oxidative stress is regarded as the initiation. However, no study investigated either the degree of endothelial DNA oxidation caused by IHG or the potential protective effects of antioxidants. In this study, DNA oxidation, including 8-hydroxy-2'-deoxyguanosine (8-OHdG) concentration and comet assay, was studied in human umbilical vein endothelial cells (HUVECs) under IHG with or without treatment of Ginkgo biloba extract (EGb 761). We found that high glucose, especially IHG, increased reactive oxygen species generation, 8-OHdG content and oxidative DNA damage in HUVECs. These high glucose-induced oxidative stress could be suppressed by EGb 761 (25-100 microg/ml) in a dose-dependent manner through the improvement of total antioxidant capacity. Our results indicated that the presence of significant DNA oxidation in HUVECs exposed to high glucose, and especially higher in the cells in IHG conditions. EGb 761, an antioxidant herbal medicine, can remarkably alleviate endothelial DNA oxidation caused by IHG, which may provide a novel approach for endothelial protection in the presence of IHG.

NOX2 deficiency ameliorates cerebral injury through reduction of complexin II-mediated glutamate excitotoxicity in experimental stroke.

Although NADPH oxidase (NOX)-mediated oxidative stress is considered one of the major mechanisms triggering the pathogenic actions of ischemic stroke and very recent studies have indicated that NADPH oxidase is a major source of reactive oxygen species (ROS) production controlling glutamate release, how neuronal NADPH oxidase activation is coupled to glutamate release is not well understood. Therefore, in this study, we used an in vivo transient middle cerebral artery occlusion model and in vitro primary cell cultures to test whether complexins, the regulators of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes necessary for vesicle fusion, are associated with NOX2-derived ROS and contribute to glutamate-mediated excitotoxicity in ischemic stroke. In this study, we first identified the upregulation of complexin II in the ischemic brain and evaluated its potential role in ischemic stroke showing that gene silencing of complexin II ameliorated cerebral injury as evidenced by reduced infarction volume, neurological deficit, and neuron necrosis accompanied by decreased glutamate levels, consistent with the results from NOX2(-/-) mice with ischemic stroke. We further demonstrated that complexin II expression was mediated by NOX2 in primary cultured neurons subjected to oxygen-glucose deprivation (OGD) and contributed to OGD-induced glutamate release and neuron necrosis via SNARE signaling. Taken together, these findings for the first time provide evidence that complexin II is a central target molecule that links NADPH oxidase-derived ROS to glutamate-mediated neuronal excitotoxicity in ischemic stroke.

Arthroscopic ankle arthrodesis in hemophilic arthropathy.

BACKGROUND: Ankle arthrodesis is an accepted treatment for patients with advanced disabling tibiotalar arthritis, mostly in osteoarthritis, rheumatoid, and posttraumatic arthritis. No detailed reports have been published regarding the use of arthroscopy for the treatment of the end-stage hemophilic ankle. The purpose of this article is to report the results of arthroscopic ankle arthrodesis in hemophilic arthropathy of the ankle. METHODS: Ten patients (10 ankle joints) who underwent arthroscopically assisted ankle arthrodesis for the treatment of end-stage hemophilic A arthritis were enrolled in this study. The rate of ankle fusion, incidence of complications, and clinical rating by the Morgan system were analyzed. RESULTS: In this series, the fusion rate was 100%, and patients achieved bone fusion as shown by radiographs. The average time to fusion was 10.5 weeks. Superficial wound infection occurred in 1 patient. According to the Morgan system, there were 8 (80%) good to excellent results and 2 (20%) fair results. All patients were satisfied with the outcome of the operation. CONCLUSIONS: Arthroscopic ankle arthrodesis was an effective alternative to open technique with established advantages in hemophilic arthropathy. LEVEL OF CLINICAL EVIDENCE: Level IV, retrospective case series.

Aggrecanases in the human synovial fluid at different stages of osteoarthritis.

In vivo and in vitro aggrecanases degrade proteoglycan aggrecan in articular cartilage. However, the expression of aggrecanases in patients in different stages of osteoarthritis (OA) has not been investigated. This study detected the expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) and ADAMTS-5 and their proteolytic products, ARGxx, in the synovial fluid (SF) of patients in different stages of OA. This study aimed to evaluate the expression of aggrecanases and to explore the respective roles of these enzymes in human cartilage degradation. A total of 144 patients with knee OA were divided into early-, middle-, and late-stage OA groups according to the degree of cartilage degradation using Recht's MRI grading standard and the modified Outerbridge classification system. Expression levels of ADAMTS-4, ADAMTS-5, and ARGxx in the SF from these patients were measured using enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Our findings showed that ADAMTS-4 and ARGxx expression levels in the early-stage group were significantly higher than in the other two groups. ADAMTS-5 in the early-stage group and ADAMTS-4, ADAMTS-5, and ARGxx in the late-stage group were significantly higher than those in the middle-stage OA group. Both ADAMTS-4 and ADAMTS-5 levels were correlated with ARGxx levels (P < 0.05). The correlation coefficients of ADAMTS-4 and ADAMTS-5 were 0.236 and 0.068, 0.729 and 0.479, and 0.675 and 0.257 in the early-, middle-, and late-stage groups, respectively, and 0.530 and 0.258 in the total SF samples. Western blot analysis revealed that the ADAMTS-4 and ADAMTS-5 in SF were 50 kDa proteins and that ARGxx in SF had at least two molecular masses, 55 kDa and 70 kDa. The expression levels of all three proteins were consistent with the ELISA results. These results suggested that aggrecanases were involved in all stages of human OA aggrecan degradation, especially in the early and late stages. ADAMTS-4 levels were higher in early- compared with middle- or late-stage OA and were also more correlated with ARGxx than ADAMTS-5; thus, ADAMTS-4 might be the principal aggrecanase of aggrecan degradation in human OA.