The lncRNA MCF2L-AS1 controls osteogenic differentiation by regulating miR-33a.

Bone homeostasis is maintained by balanced osteoblast-mediated tissue production and osteoclast-mediated tissue destruction, and is disrupted in pathological conditions such as osteoporosis. The mechanisms underlying osteogenic differentiation of bone marrow mesenchymal stem cells, which is critical to bone homeostasis, are not completely clear, despite extensively studies. Long noncoding RNAs (lncRNAs) have recently emerged as novel therapeutic targets in various diseases. However, the expression pattern and biological function of lncRNAs in osteogenic differentiation remain unclear. In this study, we aimed to determine the role of lncRNAs in osteogenic differentiation of human bone marrow mesenchymal stem cells. We found high lncRNA MCF2L-AS1 expression in human bone marrow mesenchymal stem cells, and used bioinformatics analysis to analyze its function. MCF2L-AS1 knockdown induced inhibition of osteoblast differentiation. Silencing of MCF2L-AS1 increased the expression of miR-33a and subsequently inhibited Runx2 expression at the post-transcriptional level. Moreover, MCF2L-AS1 directly interacted with miR-33a, and downregulation of miR-33a efficiently reversed the suppression of Runx2 induced by MCF2L-AS1 short hairpin RNA (shRNA). Thus, MCF2L-AS1 positively regulated the expression of Runx2 by sponging miR-33a, and promoted osteogenic differentiation in BMSCs. Our results indicated that the lncRNA MCF2L-AS1 plays a critical role in the osteogenic differentiation of BMSCs, and targeting lncRNA MCF2L-AS1 could be a promising strategy to promote osteogenic differentiation.

The Effect of Sulforaphane on the Activity and Mineralization of Osteoblasts under Oxidative Stress.

Sulforaphane (SFN) is considered an antioxidant agent, but the biological effects on hypoxia-treated osteoblasts remain unclear. Therefore, the aims of this study were to investigate the effects of SFN on the activity and mineralization of osteoblasts in hypoxia. Osteoblasts were treated with hypoxia with or without SFN, and apoptosis was assayed with caspase 3 Activity Assay Kit and flow cytometer. The levels of reactive oxygen species (ROS) were measured with DCFH-DA. The levels of glutathione (GSH) and glutathione disulphide were determined by the o-phthalaldehyde fluorimetric assay. Mineralization of Osteoblasts was detected by Alizarin red staining and alkaline phosphatase (ALP) staining, and the relative proteins levels were examined by Western blotting. Our results showed that SFN reduced the hypoxia-mediated apoptosis and ROS levels in osteoblasts. The utilization of SFN improved the inhibitory effect of osteoblast mineralization by hypoxia. Additionally, the effect of alleviating hypoxia by SFN will be an increase in osteoblast activity. These findings clarify the effects of SFN on hypoxia-treated osteogenesis and will help identify novel therapeutic strategies for the protection of skeletal health.

Multicentre comparative study of Z-shape elevating-pulling reduction and skull traction reduction for treatment of lower cervical locked facets.

PURPOSE: The aim of this study was to assess the clinical efficacy and safety of Z-shape elevating-pulling reduction as compared to that of conventional skull traction in the treatment of lower cervical locked facet. METHODS: Patients with cervical locked facet (n = 63) were retrospectively enrolled from four medical centers and divided into two groups according to the pre-operative reduction method used: Z-shape elevating-pulling reduction (Z-shape elevating group; n = 20) or traditional skull traction reduction (skull traction group; n = 43). RESULTS: The success rates, efficacy of reduction, and safety were compared between the two groups. The success rates were significantly better in the Z-shape elevating group than in the skull traction group: 87.5% (7/8) vs. 35.3% (6/17) for unilateral locked facet reduction (P = 0.03) and 100% (12/12) vs. 69.2% (18/26) for bilateral locked facet reduction (P = 0.04). There was no obvious change in American Spinal Injury Association (ASIA) grade after the reduction in either group. Combined surgery was necessary in 5% in the Z-shape elevating group vs. 27.9% in the skull traction group. Imaging showed that the segment angle and horizontal displacement were significantly improved after surgery in both groups, with no significant difference between the groups. Follow-up with radiography showed good recovery of the cervical spine sequence; all internal fixation sites were stable, with no loosening, prolapse, or breakage of internal fixators. CONCLUSIONS: Halo vest-assisted Z-shape elevating-pulling reduction appears to be a simple, safe, and effective technique for pre-operative reduction of lower cervical locked facets.

[Experimental study on the improving effect of motor nerve babysitting on delayed nerve anastomosis].

OBJECTIVE: To observe whether the motor nerve babysitter could improve the delayed nerve anastomosis and promote the functional recovery. METHODS: Sixteen SD rats weighing 200-250 g were randomly divided into 2 groups. In group A, the left musculocutaneous nerve was transected to make the model of biceps brachii denervation and anastomosed to its proximal end 6 weeks later; In group B, the musculocutaneous nerve was transected and the distal end was coapted to the purely motor medial pectoral nerve immediately (nerve babysitter) and the musculocutaneous nerve was separated from the medial pectoral nerve, and reanastomosed to its proximal end 6 weeks later. In the animal model, the left limbs served as experimental sides, the right limbs as control sides. After 6 and 12 weeks of the second surgery, behavioral test (grooming test) was performed and the degree of the biceps brachii atrophy was observed, the latent period and the amplitude of the maximum action potentials of the biceps brachii were detected, the wet muscle weight, muscle fiber cross-section area and the activity of Na+-K+-ATPase of the biceps brachii were measured. RESULTS: After 4 weeks of the second surgery, grooming behavior was found in group B, while few grooming behavior was seen in group A till 6 weeks after the secondary surgery. After 6 weeks of the second surgery, the recovery rate of the latent period and the amplitude, the wet muscle weight, muscle fiber cross-section area and the enzymatic activity of Na+-K+-ATPase of the biceps brachii in group A was 187.25% +/- 1.97%, 46.25% +/- 4.63%, 55.14% +/- 1.99%, 49.97% +/- 1.71%, and 65.81% +/- 2.24%, respectively, which was significantly different from that in group B (155.96% +/- 3.02%, 51.21% +/- 2.13%, 74.18% +/- 1.82%, 55.05% +/- 1.64% and 71.08% +/- 1.53%, respectively, P < 0.05). After 12 weeks of the second surgery, the recovery rate of the latent period and amplitude, the wet muscle weight, muscle fiber cross-section area and the enzymatic activity of Na+-K+-ATPase of the biceps brachii in group A was 145.36% +/- 3.27%, 51.84% +/- 5.02%, 77.92% +/- 1.73%, 61.04% +/- 2.68% and 71.94% +/- 1.65%, respectively, which was significantly different from that in group B (129.83% +/- 8.36%, 75.22% +/- 2.78%, 84.51% +/- 1.34%, 78.75% +/- 3.69% and 84.86% +/- 1.81%, respectively, P < 0.05). CONCLUSION: Motor nerve babysitting could reduce muscular damage after denervation, improve the effect of delayed nerve repair and promote the functional recovery of musculocutaneous nerve.