SIRT3 mitigates intervertebral disc degeneration by delaying oxidative stress-induced senescence of nucleus pulposus cells.

Senescence of nucleus pulposus (NP) cells (NPC) is a major cause of intervertebral disc degeneration (IVDD), so delay NPC senescence may be beneficial for mitigating IVDD. We studied the effect and mechanism of silent information regulator 2 homolog 3 (SIRT3) on NPC senescence in vivo and in vitro. First, we observed SIRT3 expression in normal and degenerated NPC with immunohistochemical and immunofluorescence staining. Second, using SIRT3 lentivirus transfection, reactive oxygen species probe, senescence-associated ß-galactosidase staining, polymerase chain reaction, and western blot to observe the oxidative stress, senescence, and degeneration degree among groups. Subsequently, pretreatment with adenosine monophosphate-activated protein kinase (AMPK) agonists and inhibitors, observing oxidative stress, senescence, and degeneration degree among groups. Finally, the IVDD model was constructed and divided into Ctrl, Vehicle, LV-shSIRT3, and LV-SIRT3 groups. X-ray and magnetic resonance imaging scans were performed on rat's tails after 1 week; hematoxylin and eosin and safranin-O staining were used to evaluate the degree of IVDD; immunofluorescence staining was used to observe SIRT3 expression; immunohistochemical staining was used to observe oxidative stress, senescence, and degeneration degree of NP. We found that SIRT3 expression is reduced in degenerated NP tissues but increased in H2 O2 -induced NPC. Moreover, SIRT3 upregulation decreased oxidative stress, delayed senescence, and degeneration of NPC. In addition, activation of the AMPK/PGC-1α pathway can partially mitigate the NPC oxidative stress, senescence, and degeneration caused by SIRT3 knockdown. The study in vivo revealed that local SIRT3 overexpression can significantly reduce oxidative stress and ECM degradation of NPC, delay NPC senescence, thereby mitigating IVDD. In summary, SIRT3 mediated by the AMPK/PGC-1α pathway mitigates IVDD by delaying oxidative stress-induced NPC senescence.

The biomechanical effects of graded upper articular process arthroplasty on lumbar spine: A finite element study.

BACKGROUND: Superior articular process arthroplasty is important for intervertebral foramen microscopy but may lead to spinal instability. Currently, there has been no relevant study in relation to the biomechanical analysis of superior articular process arthroplasty. Hence, this study is intended to verify biomechanical effects after unilateral S1 superior articular process arthroplasty. METHODS: Eight finite element (FE) models of lumbosacral vertebrae (L4-S) were constructed, and the superior articular process formation was simulated with the help of Geomagic studio. Then, the models were imported into Nastran software after optimization. Normal load and appropriate torque were applied to simulate forward flexion, back extension, lateral flexion and lateral rotation. In the end, changes of lumbar range of motion (ROM) and structural stress were compared with those of normal model. RESULTS: Compared with the normal model, formed from ventral to dorsal (Longitudinal), the larger motion of lumbar spine and the greater larger stress of articular process showed statistical significance (P < 0.05) in most of directions when the forming range was greater than 3/5. Formed from the apex to the base (transverse), the larger motion of lumbar spine and the greater stress of articular process showed statistical significance (P < 0.05) in most of directions when the forming range was great than 1/5. CONCLUSION: When conducting unilateral S1 articular process arthroplasty from ventral to dorsal, the forming range is recommended to be less than 3/5 of the superior articular process. Notably, it is not advisable to form from the apex to the base.

Research in the precaution of recombinant human erythropoietin to steroid-induced osteonecrosis of the rat femoral head.

Objective To elucidate the effects of recombinant human erythropoietin (rHuEPO) on steroid-induced osteonecrosis of the femoral head in rats. Methods Twenty-four adult Wistar rats were randomly divided into three groups of eight rats each. The rats in the positive control group were injected with dexamethasone at 1 mg/kg twice a week for 5 weeks. The rats in the negative control group were injected with sodium chloride alone. The rats in the experimental group were injected with dexamethasone at 1 mg/kg twice a week for 5 weeks and rHuEPO (500 u/d/kg) daily for 5 weeks. The femoral head on one side was examined by hematoxylin and eosin staining, and that on the other side was examined by CD31 staining of the capillaries. Results Hematoxylin and eosin staining in the positive control group showed that the bony trabeculae had become obviously narrow and sparse with discontinuity of the integrity. The integrity of the trabeculae was better in the experimental group than positive control group. The CD31 expression was lower in the positive control group than in the other two groups. Conclusion rHuEPO can effectively prevent osteocyte apoptosis, delaying or decreasing osteonecrosis of the femoral head.

Application of a novel design paradigm to generate general nonpeptide combinatorial templates mimicking beta-turns: synthesis of ligands for melanocortin receptors.

We report the further application of a novel approach to template and ligand design by the synthesis of agonists of the melanocortin receptor. This design method uses the conserved structural data from the three-dimensional conformations of beta-turn peptides to design rigid nonpeptide templates that mimic the orientation of the main chain C-alpha atoms in a peptide beta-turn. We report details on a new synthesis of derivatives of template 1 that are useful for the synthesis of exploratory libraries. The utility of this technique is further exemplified by several iterative rounds of high-throughput synthesis and screening, which result in new partially optimized nonpeptide agonists for several melanocortin receptors.

Oxidative induction of beta-turn conformations in cyclic peptidomimetics: conformational analyses as indicators of configuration.

Solid-phase syntheses of the cyclic peptidomimetics 1-4 were performed. Sulfide 1 and sulfoxide 3 did not tend to exist in beta-turn conformations in solution, but the sulfone 2 and the epimeric sulfoxide 4 did. The assignment of configuration of the sulfoxides is based on the conformational analyses.