ß-Ecdysone attenuates cartilage damage in a mouse model of collagenase-induced osteoarthritis via mediating FOXO1/ADAMTS-4/5 signaling axis.

BACKGROUND: ß-Ecdysone has been reported to perform a protective effect to prevent interleukin 1ß (IL-1ß)-induced apoptosis and inflammatory response in chondrocytes. In our study, the chondroprotective effects of ß-Ecdysone were explored in a mouse model of collagenase-induced osteoarthritis (OA). METHODS: Injection of collagenase in the left knee was implemented to establish a mouse model of OA. The histomorphological analysis was detected using safranine O staining. Serum pro-inflammatory cytokines were measured by ELISA assays. Protein expression in the femur and chondrocytes was analyzed using western blot. Chondrocyte apoptosis was evaluated by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. RESULTS: Treatment of OA mice with ß-Ecdysone supplementation significantly inhibited the production of pro-inflammatory cytokines. Histologic examination exhibited that the degradation of proteoglycans and the loss of trabecular bone were observed in collagenase-injected mice. However, OA-like changes were attenuated by ß-Ecdysone administration in collagenase-injected mice. Both in vivo and in vitro models, nuclear forkhead box O1 (FOXO1) protein expression was significantly reduced in the femur of collagenase-treated mice and IL-1ß-stimulated chondrocytes. However, ß-Ecdysone treatment was able to rescue FOXO1 protein expression in the nucleus to inhibit the transcription and translation of a disintegrin-like and metallopeptidase (reprolysin type) with thrombospondin type 1 motif, 4 (ADAMTS-4) and ADAMTS-5. CONCLUSION: The findings suggested that ß-Ecdysone functioned as a FOXO1 activator to protect collagenase-induced cartilage damage. FOXO1 might be a potential molecular target of ß-Ecdysone for the effective prevention and treatment of OA.

[Short term effect of total hip arthroplasty through direct anterior approach for the treatment of ankylosing spondylitis with hip flexion deformity].

OBJECTIVE: To investigate the short term clinical efficacy of direct anterior approach(DAA) total hip arthroplasty for the treatment of ankylosing spondylitis with hip flexion deformity. METHODS: From September 2014 to June 2017, 15 cases of ankylosing spondylitis with flexion deformity of the hip were treated with total hip arthroplasty through DAA approach including 12 males(17 hips) and 3 females(4 hips) with an average age of 34.4 years old ranging from 21 to 57 years old. Harris score system was used before and after operation to evaluate hip function, total hip activity and visual analogue scale (VAS) were used to evaluate the clinical efficacy. RESULTS: All 15 patients were followed up for an average of 26.2 months. In the operation, 1 case of great trochanter avulsion was given wire binding, and 1 case of linear split of the femur were given by wire binding. There were no hematoma, nerve injury and deep vein thrombosis of lower extremity. No prosthesis loosening and sinking were observed in the follow-up of X-ray film after operation. There was no heterotopic ossification after operation. After operation, 18 hips pain were relieved completely, and 3 hips pain were found when walking, which all satisfied with the daily life self-care requirements. Harris hip score, total hip motion and VAS score at 1 week after operation were significantly different from those before operation(P<0.05). There was no significant difference in the scores of HHS, total hip motion and VAS at 1, 6 months after operation(P>0.05). At the final follow-up, the Harris score was 91.2±5.3, the total hip mobility was (217.1±29.7)°, and the postoperative VAS pain score was 1.2±0.5, which was significantly different from the preoperative score(P<0.05). CONCLUSIONS: DAA approach THA has good effect in treating AS hip nonfunctional ankylosis with less trauma, less pain and quick recovery. It has a good short term effect, which can effectively improve the quality of life of patients.

Overexpression of miR­214 promotes the progression of human osteosarcoma by regulating the Wnt/ß­catenin signaling pathway.

The aberrant expression of microRNA (miR)­214 contributes to the regulation of normal and cancer cell biology, and is associated with human malignancies, however, it can operate in a contradictory manner. The role of miR­214 in osteosarcoma remains to be fully elucidated. The aim of the present study was to investigate the effects of miR­214 on osteosarcoma progression and tumor cell proliferation, and examine the molecular mechanism underlying osteosarcoma. The level of miR­214 was determined using reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analysis in osteosarcoma and matched paracancerous tissues, and in human osteosarcoma cancer cell lines. The roles of miR­214 in cell proliferation, survival and cell cycle were analyzed using miR­214 lentivirus (LV­miR­214)­infected osteosarcoma cells. In addition, the downstream target proteins in the Wnt/ß­catenin signaling pathway were evaluated using western blot analysis in the LV­miR­214­infected cells. The LV­miR­214­infected MG63 cells were also treated with exogenous ß­catenin for 24, 48 and 72 h, respectively, following which the expression of ß­catenin was measured using western blot analysis and survival was determined using a 3­(4,5­cimethylthiazol­2­yl)­2,5­diphenyl tetrazolium bromide (MTT) assay. The results of the RT­qPCR analysis showed that the expression level of miR­214 was significantly higher in the osteosarcoma tissues, compared with that in the matched paracancerous tissues, and the same was observed in the osteosarcoma cell lines. The MG63, Saos­2 and U2OS cells were infected with the hsa­mir­214 lentivirus for 48 h, and the levels of miR­214 were significantly upregulated in the human osteosarcoma cancer cells. The overexpression of miR­214 in the MG­63 and Saos­2 cells promoted cell growth, and treatment of the cells with specific antisense­microRNA oligonucleotides (AMOs) for miR­214 for indicated durations reversed the effects of miR­214. Additionally, the AMO­treated MG63 cells showed G0/G1 phase arrest, suggesting that miR­214 contributed to regulation of the cell cycle. In addition, the results of western blot analysis showed that, in the miR­214 lentivirus­infected cells, the levels of cyclin­D1, c­myc and lymphoid enhancer­binding factor­1 were significantly increased, compared with those in the control lentivirus­infected cancer cells. Of note, infection with the miR­214 lentivirus did not affect the levels of Wnt1, Wnt2, Wnt4, Axin or glycogen synthase kinase ß in the U2OS cells, whereas the expression levels of ß­catenin in the MG63 cells and Saos­2 cells were significantly increased. The addition of exogenous ß­catenin effectively reversed the efficiency of miR­214­specific AMOs, which was detected using an MTT assay. These data suggested the critical role of miR­214 in human osteosarcoma via regulation of the Wnt/ß­catenin signaling pathway and demonstrated that miR­214 is as an oncogene for human osteosarcoma.