A Smart Nanoreactor Based on an O2-Economized Dual Energy Inhibition Strategy Armed with Dual Multi-stimuli-Responsive "Doorkeepers" for Enhanced CDT/PTT of Rheumatoid Arthritis.

Activated fibroblast-like synovial (FLS) cells are regarded as an important target for rheumatoid arthritis (RA) treatment via starvation therapy mediated by glucose oxidase (GOx). However, the hypoxic RA-FLS environment greatly reduces the oxidation process of glucose and leads to a poor therapeutic effect of the GOx-based starvation therapy. In this work, we designed a hollow mesoporous copper sulfide nanoparticles (CuS NPs)-based smart GOx/atovaquone (ATO) codelivery system (named as V-HAGC) targeting RA-FLS cells to realize a O2-economized dual energy inhibition strategy to solve the limitation of GOx-based starvation therapy. V-HAGC armed with dual multi-stimuli-responsive "doorkeepers" can guard drugs intelligently. Once under the stimulation of photothermal and acidic conditions at the targeted area, the dual intelligent responsive "doors" would orderly open to realize the controllable release of drugs. Besides, the efficacy of V-HAGC would be much improved by the additional chemodynamic therapy (CDT) and photothermal therapy (PTT) stimulated by CuS NPs. Meanwhile, the upregulated H2O2 and acid levels by starvation therapy would promote the Fenton-like reaction of CuS NPs under O2-economized dual energy inhibition, which could enhance the PTT and CDT efficacy as well. In vitro and in vivo evaluations revealed V-HAGC with much improved efficacy of this combination therapy for RA. In general, the smart V-HAGC based on the O2-economized dual energy inhibition strategy combined with enhanced CDT and PTT has the potential to be an alternative methodology in the treatment of RA.

IL-10 inhibits osteoclast differentiation and osteolysis through MEG3/IRF8 pathway.

OBJECTIVE: Osteolysis caused by wear particles is the main reason for joint replacement failure. Inhibition of osteoclast differentiation relieves wear particle-induced osteolysis. Our study aimed to explore the effect of lncRNA maternally expressed gene 3 (MEG3) on osteoclast differentiation and wear particle-induced osteolysis, and to improve the potential mechanism of interleukin-10 (IL-10) inhibition on osteoclast differentiation. METHODS: Polymethylmethacrylate (PMMA) -induced osteolysis mice model and receptor activator of nuclear factor-B ligand (RANKL) -induced osteoclast differentiation model were constructed. Tartrate-resistant acidic phosphatase (TRAP) staining, hematoxylin-eosin (HE) staining, immunohistochemical staining, bone resorption assay, dual-luciferase assay, RNA pull-down assay, RNA immunoprecipitation, and chromatin immunoprecipitation were executed. RESULTS: MEG3 levels were increased and interferon regulatory factor 8 (IRF8) levels were decreased in PMMA-induced osteolysis mice. IL-10 inhibited RANKL-induced osteoclast differentiation, promoted MEG3 methylation, and inhibited MEG3 expression. Moreover, knockdown of MEG3 inhibited osteoclast differentiation and increased IRF8 levels. Meanwhile, MEG3 combined with signal transducer and activator of transcription 1 (STAT1), STAT1 combined with IRF8, and overexpression of MEG3 inhibited STAT1 binding to IRF8. Further studies have shown that knockdown of MEG3 inhibited osteoclast differentiation and alleviated osteolysis, but knockdown of IRF8 weakened these results. CONCLUSION: MEG3 regulated the expression of IRF8 by binding to STAT1, thereby affecting osteoclast differentiation and wear particle-induced osteolysis. IL-10 might inhibit osteoclast differentiation by MEG3/IRF8.

miR-34a carried by adipocyte exosomes inhibits the polarization of M1 macrophages in mouse osteolysis model.

OBJECTIVE: After bone prosthesis replacement, M1-type macrophage polarization can be induced by titanium (Ti) particles and produce inflammatory, leading to osteolysis. Adipocyte-derived exosomes (ADEs) exert immune-modulatory impact on the macrophage, while whether it can inhibit the macrophage polarization induced by Ti is unclear. This study focuses on the M1-type macrophage and aims to determine the effect of ADEs on Ti-induced M1-type macrophage polarization in osteolytic mice and the involved mechanism. METHODS: Ti particle-induced osteolysis mouse model was established and macrophages were isolated from the osteolysis site. The levels of NLRP3 and specific markers for M1-type macrophage were determined. ADEs isolated from adipocyte cell line 3T3-L1, or conditioned ADEs with low-expressed miR-34a isolated from 3T3-L1 transfected with miR-34a inhibitor were co-cultured with RAW 264.7 to determine their impact on the polarization of macrophage. RESULTS: ADEs reduced the M1-type macrophage polarization and caused the upregulation of miR-34a in macrophage of the osteolysis site of the osteolysis mouse model. Also, the level of miR-34a in ADEs was higher than that in the adipocyte. The conditioned ADEs expressed a low level of miR-34a and boosted the Ti-induced M1-type polarization. MiR-34a could target NLRP3 and negatively regulated its expression. Moreover, NLRP3 knockdown in macrophage restricted the conditioned ADEs to promote macrophage towards to Ti-induced M1-type polarization. The inhibitory function of ADEs on M1-type macrophage polarization was abolished by miR-34a silencing in the mouse osteolysis model. CONCLUSION: The miR-34a carried by ADEs reduced the polarization of M1-type macrophages by targeting macrophage NLRP3 during Ti particle-induced osteolysis.

Over-expression of miR-411-5p and miR-434-3p promotes the osteoblast differentiation by targeting GATA4.

OBJECTIVE: To investigate the role of miR-411-5p and miR-434-3p in osteoblast differentiation in particulate-induced osteolysis. METHODS: A mouse model of osteolysis and an in vitro osteolysis model were constructed. The expressions of molecules were detected using qRT-PCR and western blot. Alkaline phosphatase (ALP) activity was measured using the ALP Assay Kit, and the bone mineralization was measured using alizarin red staining. RESULTS: The expression of miR-411-5p and miR-434-3p was decreased in osteolysis mice and UHMWPE-induced mMSCs, while GATA4 protein expression was increased. Over-expression of miR-411-5p and miR-434-3p up-regulated the expressions of osteoblast gene markers, enhanced the ALP activity, promoted the bone mineralization of mesenchymal stem cells. In addition, miR-411-5p and miR-434-3p could target GATA4, and miR-411-5p/434-3p affected the expressions of osteoblast gene markers through GATA4 in vitro and in vivo. CONCLUSION: Overexpression of miR-411-5p and miR-434-3p promoted the osteoblast differentiation by inhibiting GATA4 expression.

NF-κB/let-7f-5p/IL-10 pathway involves in wear particle-induced osteolysis by inducing M1 macrophage polarization.

NF-κB signaling pathway shows significant influence on wear particle-induced osteolysis, and this study aims to explore the underlying mechanism and the role of let-7f-5p in this process. A mouse calvarial osteolysis model was constructed with PMMA particles, and the bone marrow-derived macrophages (BMMs) were isolated from the osteolysis area. The expression of miRNA and protein was determined by qRT-PCR and western blot, respectively. The level of cytokines was evaluated with ELISA. Recombinant plasmids were transfected into cells for the endogenous expression of related genes. Dual-luciferase reporter assay was performed to determine the interaction between let-7f-5p and IL-10 in macrophage RAW264.7 cells. M1 macrophage polarization and expression of let-7f-5p were promoted in BMMs of osteolysis mouse model, compared with that in sham group. The expression of let-7f-5p was increased in the process of M1 macrophage polarization that induced by PMMA. Let-7f-5p was involved in M1 polarization in macrophages that treated with PMMA. IL-10 was negatively regulated by let-7f-5p. NF-κB regulated the expression of IL-10 through let-7f-5p. NF-κB participated in the PMMA-induced M1 macrophage polarization through let-7f-5p. Let-7f-5p contributed to PMMA-induced osteolysis by promoting M1 polarization of macrophages. The NF-κB/let-7f-5p/IL-10 pathway induces M1 macrophage polarization, and thus contributing to wear particle-induced osteolysis.

LncRNA KCNQ1OT1 promotes osteogenic differentiation to relieve osteolysis via Wnt/ß-catenin activation.

BACKGROUND: Resveratrol (RSV) has been reported to stimulate osteoblast differentiation in which Wnt/ß-catenin signaling pathway played a crucial role. However, whether and how RSV activated Wnt/ß-catenin pathway in osteogenic differentiation still remained elusive. METHODS: In vivo polymethylmethacrylate (PMMA) particle-induced osteolysis (PIO) mouse model and in vitro PMMA particle-stimulated mouse mesenchymal stem cells (mMSCs) experiments were established. Relative expression levels of lncRNA KCNQ1OT1, ß-catenin, Runx2, Osterix and osteocalcin were determined using quantitative Real-Time PCR. Western blotting was used to measure ß-catenin protein expression. In addition, the alkaline phosphatase activity and mineral deposition level using alizarin red S staining were performed to examine osteogenic differentiation status. The interaction between KCNQ1OT1 and ß-catenin was confirmed by RNA pull down assay. RESULTS: RSV significantly attenuated PIO in vivo and PMMA-particle inhibition of osteogenic differentiation of mMSCs. Moreover, KCNQ1OT1 exerted the similar function in mMSCs by regulating ß-catenin. Further study demonstrated that RSV exerted its effect on osteoblastic differentiation by regulating KCNQ1OT1. Consequently, RSV alleviated PMMA-particle inhibition of osteoblastic differentiation via Wnt/ß-catenin pathway activation in vivo and in vitro. CONCLUSION: RSV accelerated osteoblast differentiation by regulating lncRNA KCNQ1OT1 via Wnt/ß-catenin pathway activation, indicating the functional role of RSV in modulating osteogenesis.

LncRNA KCNQ1OT1 ameliorates particle-induced osteolysis through inducing macrophage polarization by inhibiting miR-21a-5p.

This study aimed to investigate the mechanism of lncRNA-KCNQ1OT1 on macrophage polarization to ameliorate particle-induced osteolysis. We used polymethylmethacrylate (PMMA) to induce primary bone marrow-derived macrophages (BMMs) obtained from mice and the RAW264.7 cell line, and found that the tumor necrosis factor-alpha (TNF-α) concentration and inducible nitric oxide synthase (iNOS) expression was increased, while interleukin (IL)-10 concentration and Arg1 expression were decreased in PMMA-induced cells. KCNQ1OT1 and IL-10 expression were both suppressed and miR-21a-5p expression was promoted in PMMA-induced cells. Overexpression of KCNQ1OT1 reversed the effect of PMMA on RAW264.7 cells, such as the reduced TNF-α concentration and iNOS expression, and increased IL-10 concentration and Arg1 expression in PMMA-induced cell transfected with pcDNA-KCNQ1OT1. The luciferase assay confirmed that IL-10 is a target of miR-21a-5p. RNA immunoprecipitation (RIP) and RNA pull-down experiments demonstrated that KCNQ1OT1 functions as a miR-21a-5p decoy. Thus, lncRNA KCNQ1OT1 induces M2 macrophage polarization to ameliorate particle-induced osteolysis by inhibiting miR-21a-5p.

The effect of vitamin D supplementation on knee osteoarthritis: A meta-analysis of randomized controlled trials.

OBJECTIVE: We conducted a meta-analysis of RCTs to evaluate the effects of vitamin D supplementation in the prevention of symptom and structural progression of knee OA. METHODS: PubMed, Embase, and Web of Science databases were searched to identify relevant studies. Outcomes included Western Ontario and McMaster Universities Arthritis Index (WOMAC) pain, function, stiffness, tibial cartilage volume, and serum vitamin D3 levels, and adverse events. Results were expressed as weight mean difference (WMD) with 95% confidence interval (CI), and risk ratio (RR) with 95%CI. RESULTS: Four RCTs involving 1136 patients were included in this study. Pooled estimates suggested that vitamin D supplementation was associated with a significant reduction in WOMAC pain, and WOMAC function, but not in WOMAC stiffness. Vitamin D supplementation increased the serum vitamin D3 level, but had no effect on tibial cartilage volume. Subgroup analysis showed that, a daily supplement of more than 2000 IU vitamin D significantly decreased the WOMAC pain and WOMAC function. There was no significant difference in incidence of adverse events between the vitamin D and placebo groups. CONCLUSION: Vitamin D supplementation was effective in improving the WOMAC pain and function in patients with knee OA. However, it had no beneficial effect on the prevention of tibial cartilage loss. Therefore, there is currently a lack of evidence to support the use of vitamin D supplementation in preventing the progression of knee OA.

Down-regulation of RBP-J mediated by microRNA-133a suppresses dendritic cells and functions as a potential tumor suppressor in osteosarcoma.

BACKGROUND AND OBJECTIVE: In recent years, immunotherapy for the treatment of tumors have been established. Dendritic cells (DCs) are extremely efficient and professional antigen presenting cells (APCs), which are an important target for immune therapeutic interventions in cancer. In present study, we investigated whether RBP-J signaling regulated by miR-133a was involved in the DCs mediated tumor suppressor in osteosarcoma. METHODS: DCs were isolated from 30 osteosarcoma patients and 30 healthy subjects. Mouse macrophage-like cell line RAW264.7 were cultured and osteosarcoma mouse model with injection of murine osteosarcoma cell line S180 were established. RESULTS: In osteosarcoma patients, miR-133a expression level of DCs was increased, and RBP-J expression in mRNA and protein levels were decreased. MiR-133a inhibitor promoted maturation and activation of DCs in osteosarcoma patients. In osteosarcoma mouse model, miR-133a mimic suppressed the maturation and activation of spleen DCs, while miR-133a inhibitor promoted them. Overexpression of miR-133a decreased therapeutic effect of DCs on osteosarcoma mice. In RAW264.7 cells, miR-133a was observed to target RBP-J and regulate its expression. MiR-133a mimic inhibited the maturation of DCs in cells exposed to LPS, the effect of which was reversed by overexpression of RBP-J. CONCLUSION: RBP-J mediated by miR-133a probably contributed to the regulation of DCs maturation and activation in osteosarcoma, which functioned as a therapeutic target for the immunotherapy in cancers.

Periarticular multimodal drug injection in total knee arthroplasty.

PURPOSE: A systematic review and meta-analysis based on randomized controlled trials (RCTs) were conducted to evaluate the efficiency and safety of periarticular multimodal drug injection in total knee arthroplasty (TKA). METHODS: Periarticular injection with the use of multimodal drugs is an efficient alternative for postoperative analgesia in TKA. A systematical electronic search was performed to identify the eligible RCTs in the databases of PubMed, Embase, Cochrane Central Register of Controlled Trials, Web of Science and the Chinese Biomedical Literature Database. Two independent reviewers completed data collection and assessment of methodological quality. The quality of evidence of outcomes was judged using GRADE criteria. Meta-analysis was performed for the outcomes of pain, straight leg raise, operating time, hospital stay and complications. RESULTS: Ten RCTs including eight studies with 1,216 TKAs in 835 patients met the inclusion criteria. Periarticular injection with multimodal drugs in TKA was associated with short-term benefits in terms of pain relief, straight leg raise, narcotic consumption, and the rates of nausea, vomiting, rash and pruritus. There were no statistically significant differences in operating time, hospital stay, wound complications and deep vein thrombosis between both groups. CONCLUSIONS: The current evidence suggests that periarticular multimodal drug injection in TKA provides short-term advantages in pain relief, straight leg raise and postoperative complications.

Subvastus versus medial parapatellar approach in total knee arthroplasty: meta-analysis.

The subvastus and medial parapatellar approaches are 2 commonly performed techniques in total knee arthroplasty, but the optimal approach for total knee arthroplasty remains controversial. The purpose of this study was to compare the effectiveness and safety of the subvastus vs medial parapatellar approach.The PubMed, Embase, Cochrane Library, Inter-Services Intelligence Web of Knowledge, and Chinese Biomedical Literature databases were searched for eligible quasi-randomized, controlled and randomized, controlled trials. Two authors independently extracted data and assessed the methodological quality of the included studies according to the Cochrane handbook version 5.1.0. Statistical analysis was performed using Review Manager version 5.1 software. Eight randomized, controlled trials and 1 quasi-randomized, controlled trial involving 940 primary total knee arthroplasties were included for meta-analysis. Meta-analysis revealed significant differences favoring the subvastus group in Knee Society Score in terms of function at 4 to 6 weeks (weighted mean difference [WMD]=5.09; 95% confidence interval [CI], 3.08 to 7.09; P<.01) and knee score at 12 months (WMD=2.17; 95% CI, 0.01 to 4.34; P=.05) and lateral retinacular release (odds ratio=0.34; 95% CI, 0.14 to 0.79; P=.01) when compared with the medial parapatellar approach. However, both groups showed similar results in range of motion (P>.05), operative time (WMD=2.15; 95% CI, -3.61 to 7.35; P=.42), blood loss (WMD= -31.07; 95% CI, -91.89 to 29.75; P=.32), hospital stay (WMD= -0.18; 95% CI, -0.67 to 0.31; P=.47), and postoperative complications (P>.05).