Osteoclasts (OCs) are multinucleated cells that play a pivotal role in skeletal development and bone remodeling. Abnormal activation of OCs contributes to the development of bone-related diseases, such as osteoporosis, bone metastasis and osteoarthritis. Restoring the normal function of OCs is crucial for bone homeostasis. Recently, RNA therapeutics emerged as a new field of research for osteoarticular diseases. The aim of this study is to use non-coding RNAs (ncRNAs) to molecularly engineer OCs and modulate their function. Specifically, we investigated the role of the microRNAs (namely miR-16) and long ncRNAs (namely DLEU1) in OCs differentiation and fusion. DLEU1/DLEU2 region, located at chromosome 13q14, also encodes miR-15 and miR-16. Our results show that levels of these ncRNA transcripts are differently expressed at distinct stages of the OCs differentiation. Specifically, silencing of DLEU1 by small interfering RNAs (siDLEU1) and overexpression of miR-16 by synthetic miRNA mimics (miR-16-mimics) led to a significant reduction in the number of OCs formed per field (OC/field), both at day 5 and 9 of the differentiation stage. Importantly, time-lapse analysis, used to track OCs behavior, revealed a significant decrease in fusion events after transfection with siDLEU1 or miR-16-mimics and an alteration in the fusion mode and partners. Next, we investigated the migration profile of these OCs, and the results show that only miR-16-mimics-OCs, but not siDLEU-OCs, have a lower percentage of immobile cells and an increase in cells with mobile regime, compared with controls. No differences in cell shape were found. Moreover, mass-spectrometry quantitative proteomic analysis revealed independent effects of siDLEU1 and miR-16-mimics at the protein levels. Importantly, DLEU1 and miR-16 act by distinct processes and pathways. Collectively, our findings support the ncRNAs DLEU1 and miR-16 as therapeutic targets to modulate early stages of OCs differentiation and, consequently, to impair OC fusion, advancing ncRNA-therapeutics for bone-related diseases.
Delayed onset muscle soreness (DOMS) in the quadriceps is frequent in runners finishing a marathon race, and may result in several days of discomfort and pain. There is an increasing clinical evidence that noninvasive, pulsed electromagnetic field therapy (PEMF) can have physiological effect on inflammation and tissue repair. The purpose of this pilot study was to investigate the effect of PEMF on quadriceps muscle soreness in marathon runners and to use the data to calculate an appropriate sample size for a subsequent study. The design was a randomized double-blind prospective study covering a 5 days period after completion of a beach marathon. After the marathon all 74 runners that completed the 42.195 km were asked to participate in the study. Forty-six agreed to enter the study and were block randomized into an intervention group or a control group. The intervention group received an active pulsed electromagnetic field device, and the control group received a sham device. The sham devices were used in exactly the same manner but produced no electromagnetic field. The active PEMF device does not produce heat or cause any sensation in the tissue allowing participants to be blinded to treatment. The pulsed electromagnetic field signals of a 2-msec burst of 27.12-MHz sinusoidal waves were repeated at two bursts per second. Peak magnetic field was 0.05 G, which induced an average electric field of 10 mV/cm in the tissue with an effect of 7.3 mW/cm3. All subjects were instructed to place the device on the most painful area of the quadriceps for 20 minutes four times a day. Pain intensity was measured three times a day with the Visual Analogue Scale (VAS) during a 90o squat with a self-administered questionnaire. Data were non-parametric and compared with a two-sample Wilcoxon rank-sum test.Introduction
Material and methods
