The Nrf2/HO-1 pathway participates in the antiapoptotic and anti-inflammatory effects of platelet-rich plasma in the treatment of osteoarthritis.

INTRODUCTION: We aimed to explore the molecular mechanisms through which platelet-rich plasma (PRP) attenuates osteoarthritis (OA)-induced pain, apoptosis, and inflammation. METHODS: An in vivo model of OA was established by injuring rats using the anterior cruciate ligament transection method, whereas an in vitro model was generated by exposing chondrocytes to interleukin (IL)-1ß. Both models were then treated with PRP. RESULTS: In both the in vivo and in vitro models, OA led to the suppression of the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway, whereas treatment with PRP reactivated this molecular axis. Inhibition of the Nrf2/HO-1 pathway using the Nrf2 inhibitor brusatol or through Nrf2 gene silencing counteracted the effects of PRP in reducing the tenderness and thermal pain thresholds of OA rats. Additionally, PRP reduced the mRNA expression of IL-1ß, IL-6, tumor necrosis factor-alpha (TNF-α), and matrix metallopeptidase 13 (MMP-13) and the protein expression of B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X-protein (Bax), and caspase-3. Furthermore, inflammation and apoptosis were induced by brusatol treatment or Nrf2 silencing. Additionally, in the in vitro model, PRP treatment increased the proliferation of chondrocytes and attenuated their inflammatory response and apoptosis, effects that were abrogated by Nrf2 depletion. CONCLUSIONS: The Nrf2/HO-1 pathway participates in the PRP-mediated attenuation of OA development by suppressing inflammation and apoptosis.

[Retracted] Hesperidin exhibits in vitro and in vivo antitumor effects in human osteosarcoma MG­63 cells and xenograft mice models via inhibition of cell migration and invasion, cell cycle arrest and induction of mitochondrial­mediated apoptosis.

[This retracts the article DOI: 10.3892/ol.2018.9439.].

Platelet-rich plasma treatment alleviates osteoarthritis-related pain, inflammation, and apoptosis by upregulating the expression levels of microRNA-375 and microRNA-337.

OBJECTIVE: The present study was designed to determine the molecular mechanism by which platelet-rich plasma (PRP) acts on Osteoarthritis (OA) -related pain, inflammation, and apoptosis in vivo and in vitro. MATERIALS AND METHODS: An in vivo OA model was established in rats using anterior cruciate ligament transection, and an in vitro OA model was created by treating chondrocytes with IL-1ß. Then, the induced rats and chondrocytes were treated with PRP. Real-time PCR were used to examine the expression of micorRNAs (miRs) and mRNAs of inflammatory cytokines. WB were performed to detect the expression of apoptotic factors and Wnt/ß-catenin signals. Structural damage of the cartilage and pain in OA rats were analyzed and represented by Mankin Score, OARSIS score, Tender threshold, and Thermal pain threshold. CCK-8 assay and flow cytometry were used to determine cell viability and apoptosis. RESULTS: The expression levels of miR-337 and miR-375 were downregulated in the in vivo and vitro OA models; however, PRP treatment elevated their levels. miR-337 and miR-375 inhibition reversed the effects of PRP of reducing tenderness and thermal pain thresholds in OA rats. Moreover, PRP decreased the mRNA expression levels of MMP-13, Bax, and inflammatory factors, such as IL-1ß, IL-18, and TNF-α, as well as increased the expression levels of collagen II and antiapoptotic Bcl-2. The decrease in inflammation and apoptosis was reversed by miR-337 and miR-375 inhibition, respectively. DISCUSSION AND CONCLUSIONS: In conclusion, miR-337 and miR-375 are involved in PRP-delayed OA progression by affecting inflammation and apoptosis.

SP1-Induced Upregulation of lncRNA LINC00514 Promotes Tumor Proliferation and Metastasis in Osteosarcoma by Regulating miR-708.

BACKGROUND: Growing studies have suggested the dysregulation of long non-coding RNAs (lncRNAs) in several tumors, including osteosarcoma (OS). However, limited studies report metastasis-associated lncRNAs in OS. Our present study aimed to explore the roles of lncRNA LINC00514 (LINC00514) in OS. MATERIALS AND METHODS: The LINC00514 expression was measured using qPCR assays in OS tissues and cell lines. The clinical significance of LINC00514 expression in OS patients was analyzed using chi-square test, Kaplan-Meier assays and multivariate analysis. The possible effects of LINC00514 in tumor cellular progression were determined using a series of functional assays. The mechanisms of LINC00514 action were explored through bioinformatics, luciferase reporter assays and RT-PCR assays. The mechanisms involved the upregulation of LINC00514 expression in OS were determined using luciferase reporter and chromatin immunoprecipitation (ChIP) assays. RESULTS: We showed that LINC00514 expressions were distinctly upregulated in both OS tissues and cell lines, especially in advanced cases. High levels of LINC0051 were positively correlated with advanced tumor stages, distant metastasis, and reduced survival of patients with OS. Functional experiments indicated that silencing of LINC00514 suppressed the ability of cell growth, colony formation and metastasis, whereas promoted cell apoptosis in vitro. Mechanistic investigation revealed that LINC00514 could directly bind to miR-708 and effectively serve as a ceRNA for miR-708. In addition, LINC00514 was upregulated by the transcription factor SP1. CONCLUSION: Our findings revealed SP1-induced upregulation of LINC00514 as an oncogene in OS through competitively binding to miR-708, suggesting that there are potential diagnostic and treatment values of LINC00514 in OS.

Hesperidin exhibits in vitro and in vivo antitumor effects in human osteosarcoma MG-63 cells and xenograft mice models via inhibition of cell migration and invasion, cell cycle arrest and induction of mitochondrial-mediated apoptosis.

The objective of the present study was to investigate the anticancer properties of hesperidin against human osteosarcoma MG-63 cells. Its effects on apoptosis, cell migration, cell invasion and cell cycle arrest, and its effects on tumor volume and weight were also evaluated in the present study. MTS assay was used to study the cytotoxic effects of the compound on cell viability. Effects on apoptosis and cell cycle arrest were evaluated by flow cytometry. In vitro wound healing assay and Matrigel assay were performed to study the effects of hesperidin on cell migration and cell invasion, respectively. Hesperidin exerted dose-dependent and time-dependent growth inhibitory effects on cervical cancer cells with IC50 values of 33.5, 23.8 and 17.6 µM, respectively, at 24, 48 and 72 h time intervals. Hesperidin led to early and late apoptosis induction in these cells. Hesperidin-treated cells also led to G2/M phase cell cycle arrest, which exhibited strong dose-dependence. Hesperidin treatment also led to inhibition of cell migration and invasion.

Bone Morphogenic Protein-2 (rhBMP2)-Loaded Silk Fibroin Scaffolds to Enhance the Osteoinductivity in Bone Tissue Engineering.

There is an increasing demand for formulations of silk fibroin (SF) scaffolds in biomedical applications. SF was crosslinked via glutaraldehyde with osteoinductive recombinant human bone morphogenic protein-2 (rhBMP2) of different ratios viz. (i) 3% SF with no rhBMP2 (SF), (ii) 3% SF with equal amount of rhBMP2 (SF+BMP2), and (iii) 12% SF with 3% of rhBMP2 (4SF+BMP2), and these solutions were used in electrospinning-based fabrication of nanoscaffolds for evaluating increased osteoinductive potential of SF scaffolds with rhBMP2. Stress-strain relationship suggested there is no loss in mechanical strength of fibers with addition of rhBMP2, and mechanical strength of scaffold was improved with increase in concentration of SF. rhBMP2 association increased the water retention capacity of scaffold as evident from swelling studies. Viability of hMSCs was found to be higher in conjugated scaffolds, and scaffolds do not exhibit any cytotoxicity towards guest cells. Cells were found to have higher alkaline phosphatase activity in conjugated scaffolds under in vitro and in vivo conditions which establishes the increased osteoinductivity of the novel construct. The scaffolds were found to be effective for in vivo bone formation as well.

Efficiency and safety of tranexamic acid in reducing blood loss in total shoulder arthroplasty: A systematic review and meta-analysis.

OBJECTIVE: This meta-analysis aimed to evaluate the efficiency and safety of tranexamic acid for reducing blood loss and transfusion requirements in patients undergoing total shoulder arthroplasty. METHODS: A systematic search was performed in Embase (1980-2017.04, embase.com), Medline (1966-2017.04, medline.com), PubMed (1966-2017.04, pubmed.com), ScienceDirect (1985-2017.04, sciencedirect.com), and Web of Science (1950-2017.04, webofknowledge.com). Study which assessed the efficiency and safety of tranexamic acid in total shoulder arthroplasty was selected. Meta-analysis was performed using Stata 11.0 software. RESULTS: In all, 484 patients from 2 randomized controlled trials (RCTs) and 2 non-RCTs were subjected to meta-analysis. The present meta-analysis demonstrated that there was less total blood loss (mean difference [MD] -172.16, 95% confidence interval [CI] -35.46 to -308.87, P = .01, d = 0.33) and transfusion rate (odds ratio 0.34, 95% CI 0.13 to 0.91, P = .03, d = 0.29) in tranexamic acid groups compared with the control groups. There were no significant differences in duration of surgery (MD 0.02, 95% CI -0.12 to 0.22, P = .89, d = 0.19), length of stay (MD -0.06, 95% CI -0.26 to 0.14, P = .56, d = 0.20), or incidence of adverse effects such as deep venous thrombosis (odds ratio 1.15, 95% CI 0.33 to 4.00, P = .83, d = 0.53). CONCLUSION: Clinical application of tranexamic acid seemed to result in significant reductions in total blood loss, hemoglobin decline and transfusion requirements following total shoulder arthroplasty. Moreover, no increased risk of the thrombotic events was identified. Due to the limited quality of the evidence currently available, higher quality RCTs are required.

Low-frequency vibration treatment of bone marrow stromal cells induces bone repair in vivo.

OBJECTIVES: To study the effect of low-frequency vibration on bone marrow stromal cell differentiation and potential bone repair in vivo. MATERIALS AND METHODS: Forty New Zealand rabbits were randomly divided into five groups with eight rabbits in each group. For each group, bone defects were generated in the left humerus of four rabbits, and in the right humerus of the other four rabbits. To test differentiation, bones were isolated and demineralized, supplemented with bone marrow stromal cells, and implanted into humerus bone defects. Varying frequencies of vibration (0, 12.5, 25, 50, and 100 Hz) were applied to each group for 30 min each day for four weeks. When the bone defects integrated, they were then removed for histological examination. mRNA transcript levels of runt-related transcription factor 2, osteoprotegerin, receptor activator of nuclear factor κ-B ligan, and pre-collagen type 1 α were measured. RESULTS: Humeri implanted with bone marrow stromal cells displayed elevated callus levels and wider, more prevalent, and denser trabeculae following treatment at 25 and 50 Hz. The mRNA levels of runt-related transcription factor 2, osteoprotegerin, receptor activator of nuclear factor κ-B ligand, and pre-collagen type 1 α were also markedly higher following 25 and 50 Hz treatment. CONCLUSION: Low frequency (25-50 Hz) vibration in vivo can promote bone marrow stromal cell differentiation and repair bone injury.

[Surgical treatment for chronic achilles tendon rupture and severe scarring].

OBJECTIVE: To evaluate the clinical efficacy of autologous semitendinosus and gracilis tendon grafting with anchor repair for the treatment of chronic achilles tendon rupture and severe scarring. METHODS: From April 2010 to October 2012,26 patients with chronic achilles tendon rupture(with Myerson type III ) and severe scarring were treated with autologous semitendinosus and gracilis tendon grafting with anchor repair. There were 19 males and 7 females,with an average age of 32 years old (ranged, 22 to 47 years). The time from injury to surgery was from 3 to 12 months (7 months on average). The plantar flexion strength of all injuried feet attenuated and single heel rise test were positive in 26 cases before operation. Plaster immobilization and routine rehabilitation therapy were performed after operation. Clinical effects were evaluated by Arner-lindholm criterion and complications were observed after operation. RESULTS: All the patients were followed up from 12 to 24 months with a mean of 16 months. No complications such as achilles tendon re-rupture, wound infection, etc were found during follow-up period. According to the Arner-Lindholm standard, 15 cases got excellent results and 11 good. CONCLUSION: Using autologous semitendinosus and gracilis tendon grafts with anchor repair to treat chronic achilles tendon rupture and severe scarring is a perfect surgical procedure.

[Patterns of osteoprotegerin or nuclear factor kappa B ligand gene expression in the treatment of bone defect with bone marrow stem cell transplantation and low-frequency vibration].

OBJECTIVE: To explore the effect of low frequency vibration (LFV)on the osteogenic differentiation regulating capability of bone marrow stromal cell (BMSC)and the expressions of OPG (osteoprotegerin) mRNA and RANKL (nuclear factor kappa B ligand) mRNA through living animal experiment. METHODS: Both BMSC transplantation and low-frequency vibration were employed to treat bone defects. The groups were randomized into non-vibration and vibration of different frequencies. The vibration group received vibrating interventions at Day 7 for 5 weeks. After vibrations, the BMSC OPG and RANKL mRNA of different frequency groups were detected. RESULTS: The BMSC OPG and RANKL gene expressions significantly increased (P < 0.05), especially at 25 Hz (P < 0.01). And for the vibration group at 100 Hz, the BMSC OPG and RANKL gene expressions decreased (P < 0.05). CONCLUSION: Low-frequency vibration may promote the osteogenic differentiation capability of BMSC probably through regulating the OPG/RANKL mRNA expression, directly promoting bone formation and inhibiting bone resorption.

[A study of 22 displaced intraarticular calcaneal fractures using locking plates with and without bone graft].

OBJECTIVE: To analyze locking plates with bone graft fusion in treating displaced intraarticular calcaneal fractures and determine whether it is beneficial in maintaining restoration of calcaneal height and anatomic reduction of the articular surface. METHODS: From January 2007 to January 2008, 22 patients with displaced intraarticular calcaneal fractures were treated with locking plates with and without bone graft (divided into the bone graft group and non-bone graft group). There were 17 males and 5 females, ranging in age from 18 to 59 years with the mean of 35 years. Sanders III was in 14 cases and Sanders IV in 8 cases. Autologous iliac bone filled defects with locking plates fixation for the bone graft group; just locking plates fixation were performed for non-bone graft group. The Böhler angle and Gissane angle were measured before and after operation. The foot function of two groups were compared according to Maryland standard at the 6th month, 1, 2 years after operation. RESULTS: All patients were followed up with an average of 25 months. There was no significant difference in the recovery of Böhler angle and Gissane angle between two groups (P > 0.05). After the 6 months,1, 2 years, there was no significant difference in the foot function between two groups (P > 0.05), in bone graft group, excellent result was in 6, 7 ,7 cases respectively; and in non-bone graft group, excellent results in 5, 6, 7 cases respectively. CONCLUSION: Bone graft in the surgical treatment of calcaneal fractures is not an advantage.