microRNA-29b-3p/sirtuin-1/peroxisome proliferator-activated receptor γ suppress osteogenic differentiation.

Osteoporosis is described as an age-associated impairment of bone formation. microRNA (miR)-29b-3p was thought to be linked to osteoblast differentiation; however, the underlying molecular pathways are yet unknown. The study's goal was to look into the involvement of miR-29b-3p in osteoporosis and the pathophysiological mechanisms. A murine model of estrogen deficiency-induced bone loss was established to simulate postmenopausal osteoporosis. Reverse transcription-quantitative PCR (RT-qPCR) was performed to assess the level of miR-29b-3p of bone tissue. Additionally, miR-29b-3p/sirtuin-1 (SIRT1)/peroxisome proliferator-activated receptor γ (PPARγ) axis in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was examined. Osteogenesis-related markers, including alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2), were assessed at protein and molecular levels. ALP staining and Alizarin Red staining were used to detect ALP activity and calcium deposition. The ovariectomy group was shown to express miR-29b-3p at higher levels in vitro, and miR-29b-3p mimics suppressed osteogenic differentiation and protein/mRNA expression levels of osteogenesis-related markers in vivo. SIRT1 was identified as a target of miR-29b-3p using luciferase reporter assays. Overexpression of SIRT1 reduced the inhibition of osteogenic differentiation by miR-29b-3p. Rosiglitazone, an activator of PPARγ signaling, was able to reverse the downregulation of the osteogenic differentiation of BMSCs and the protein expression of PPARγ caused by miR-29b-3p inhibitors. The results revealed that osteogenesis was suppressed by miR-29b-3p, which blocks the SIRT1/PPARγ axis. These results suggested that postmenopausal osteoporosis could be treated by targeting miR-29b-3p SIRT1/PPARγ.

Neural substrates in patients with visual-spatial neglect recovering from right-hemispheric stroke.

Visual-spatial attention disorder after stroke seriously affects recovery and quality of life in stroke patients. Previous studies have shown that some patients recovery rapidly from visual-spatial neglect (VSN), but the brain networks underlying this recovery are not well understood. Using functional magnetic resonance imaging, we aimed to identify network differences between patients who rapidly recovered from VSN and those with persistent VSN. The study included 30 patients with VSN who suffered subacute stroke. Patients were examined 2-4 weeks after stroke onset and 4 weeks after the initial assessment. At the last evaluation, patients in the persistent VSN (n = 15) and rapid recovery (n = 15) groups underwent paper-and-pencil tests. We defined the bilateral frontal eye fields, bilateral intraparietal sulcus in the dorsal attention network, and right temporoparietal junction and ventral frontal cortex areas in the ventral attention network as regions of interest (ROI) and measured whole-brain ROI-based functional connectivity (FC) and amplitude of low-frequency fluctuations (ALFF) in subacute right-hemisphere stroke patients. VSN recovery was associated with changes in the activation of multiple bilateral attentional brain regions. Specifically, persistent VSN was associated with lower FC in the right superior frontal gyrus, right inferior temporal gyrus, right medial orbitofrontal cortex, left precuneus, right inferior parietal gyrus, right medial frontal gyrus, right rectus gyrus, left superior frontal gyrus, left middle cingulate gyrus, right superior temporal pole, right postcentral gyrus, and right posterior cingulate gyrus compared to that in those with rapid recovery, whereas ALFF in the left cerebellum were decreased in patients with persistent VSN. Our results demonstrate that the DAN rather than the VAN, plays a more important role in recovery from VSN, and that the cerebellum is involved in recovery. We believe that our results supplement those of previous studies on recovery from VSN.

Therapeutic Effects of Transcranial Magnetic Stimulation on Visuospatial Neglect Revealed With Event-Related Potentials.

This study aimed to investigate changes in attention processing after low-frequency repetitive transcranial magnetic stimulation (rTMS) over the left posterior parietal cortex to better understand its role in visuospatial neglect (VSN) rehabilitation. The current study included 10 subacute stroke patients with VSN consecutively recruited from the inpatient stroke rehabilitation center at Xuanwu Hospital (the teaching hospital affiliated with Capital Medical University) between March and November 2019. All patients performed a battery of tasks (including line bisection, line cancellation, and star cancellation tests) two weeks before treatment and at the beginning and end of treatment; the attentive components of the test results were analyzed. In addition, low-frequency rTMS was used to stimulate the left posterior parietal cortex for 14 days and event-related potential data were collected before and after the stimulation. Participants were evaluated using a target-cue paradigm and pencil-paper tests. No significant differences were detected on the battery of tasks before rTMS. However, we found that rTMS treatment significantly improved the response times and accuracy rates of patients with VSN. After rTMS, the treatment side (left) amplitude of P300 following an event-related potential was higher than that before treatment (left target, p = 0.002; right target, p = 0.047). Thus, our findings suggest that rTMS may be an effective treatment for VSN. The observed increase in event-related potential amplitude supports the hypothesized compensational role of the contralesional hemisphere in terms of residual performance. Our results provide electrophysiological evidence that may help determine the mechanisms mediating the therapeutic effects of rTMS.

Autogenous bone particles combined with platelet-rich plasma can stimulate bone regeneration in rabbits.

Long-term bone defects are a key clinical problem. Autogenous bone graft remains the gold standard for the treatment of these defects; however, improving the osteogenic properties and reducing the amount of autogenous bone is challenging. Autologous platelet-rich plasma (PRP) has been widely considered for treatment, due to its potentially beneficial effect on bone regeneration and vascularization. The aim of the present study was to explore the effects of autogenous bone particles combined with PRP on repairing segmental bone defects in rabbits. Briefly, a critical-size diaphyseal radius defect was established in 45 New Zealand White rabbits. Animals were randomly divided into four groups, according to the different implants: Group A, empty bone defect; group B, PRP; group C, autogenous bone particles + bone mesenchymal stem cells (BMSCs) on the left radius; group D, autogenous bone particles + PRP + BMSCs on the right radius. Bone samples were collected and further analyzed using X-ray, histology and histomorphometry 4, 8 and 12 weeks post-surgery. In addition, the effect of PRP on cell proliferation was detected by Cell Counting Kit-8 and the concentrations of growth factors (GFs), transforming GF (TGF)-ß1 and platelet-derived GF (PDGF), in PRP were verified by ELISA. X-ray, histology and histomorphometry data revealed that the fraction area of the newly formed bone was larger in group D. In addition, PRP could improve cell proliferation, osteogenic differentiation and the release of GFs, TGF-ß1 and PDGF-AB. In conclusion, these findings indicated that an autogenous bone particle + PRP + BMSC scaffold may be used as a potential treatment strategy for segmental defects in humans.

Visual processing features in patients with visual spatial neglect recovering from right-hemispheric stroke.

OBJECTIVE: Visual spatial neglect (VSN) is a disorder of spatial-temporal attention, often as a result of traumatic brain injury, including stroke. Accumulating evidence suggests that the recovery from VSN follows a very predictable pattern. In this study, we aimed to determine the specific electrophysiology readout that might have predictive value for recovery from VSN in the typical early events, including the recovery rate of visual processing, within the first four weeks of recovery. METHODS: This was a prospective study of 18 right ischemic stroke patients with VSN who performed a visual cue-target task within 3 days after stroke. The patients were divided into two groups according to their outcome. We compared behavioral data, the amplitudes and latencies of ERP components（P1, N1, and P300） between patients with persistent-VSN (P-VSN) and those with rapid recovery-VSN (R-VSN). RESULTS: The amplitudes and latencies of the P1 and N1 components were not significantly influenced by the validity of the cue-based expectancy (all p > 0.05). However, a longer mean P300 latency evoked an effective cue (p < 0.001), and there was a significant difference between the P-VSN and R-VSN groups when using the left target (left hemisphere, p = 0.014; right hemisphere, p = 0.027). The recovery rate found in our study (18.75% at four weeks after stroke) was lower than that of previously reported studies. CONCLUSIONS: Our findings support the use of the event-related potential as a tool for investigating rapid recovery from VSN after stroke and suggest that other factors, such as an asymmetrical omission toward the contralateral side or impairment in the temporal processing capacity, might also be potential biomarkers of recovery.

Visual-spatial neglect after right-hemisphere stroke: behavioral and electrophysiological evidence.

BACKGROUND: Visual-spatial neglect (VSN) is a neuropsychological syndrome, and right-hemisphere stroke is the most common cause. The pathogenetic mechanism of VSN remains unclear. This study aimed to investigate the behavioral and event-related potential (ERP) changes in patients with or without VSN after right-hemisphere stroke. METHODS: Eleven patients with VSN with right-hemisphere stroke (VSN group) and 11 patients with non-VSN with right-hemisphere stroke (non-VSN group) were recruited along with one control group of 11 age- and gender-matched healthy participants. The visual-spatial function was evaluated using behavioral tests, and ERP examinations were performed. RESULTS: The response times in the VSN and non-VSN groups were both prolonged compared with those of normal controls (P < 0.001). In response to either valid or invalid cues in the left side, the accuracy in the VSN group was lower than that in the non-VSN group (P < 0.001), and the accuracy in the non-VSN group was lower than that in controls (P < 0.05). The P1 latency in the VSN group was significantly longer than that in the control group (F[2, 30] = 5.494, P = 0.009), and the N1 amplitude in the VSN group was significantly lower than that in the control group (F[2, 30] = 4.343, P = 0.022). When responding to right targets, the left-hemisphere P300 amplitude in the VSN group was significantly lower than that in the control group (F[2, 30] = 4.255, P = 0.025). With either left or right stimuli, the bilateral-hemisphere P300 latencies in the VSN and non-VSN groups were both significantly prolonged (all P < 0.05), while the P300 latency did not differ significantly between the VSN and non-VSN groups (all P > 0.05). CONCLUSIONS: Visual-spatial attention function is impaired after right-hemisphere stroke, and clinicians should be aware of the subclinical VSN. Our findings provide neuroelectrophysiological evidence for the lateralization of VSN.

Assessment of the degradation rates and effectiveness of different coated Mg-Zn-Ca alloy scaffolds for in vivo repair of critical-size bone defects.

Surgical repair of bone defects remains challenging, and the search for alternative procedures is ongoing. Devices made of Mg for bone repair have received much attention owing to their good biocompatibility and mechanical properties. We developed a new type of scaffold made of a Mg-Zn-Ca alloy with a shape that mimics cortical bone and can be filled with morselized bone. We evaluated its durability and efficacy in a rabbit ulna-defect model. Three types of scaffold-surface coating were evaluated: group A, no coating; group B, a 10-µm microarc oxidation coating; group C, a hydrothermal duplex composite coating; and group D, an empty-defect control. X-ray and micro-computed tomography(micro-CT) images were acquired over 12 weeks to assess ulnar repair. A mechanical stress test indicated that bone repair within each group improved significantly over time (P < 0.01). The degradation behavior of the different scaffolds was assessed by micro-CT and quantified according to the amount of hydrogen gas generated; these measurements indicated that the group C scaffold better resisted corrosion than did the other scaffold types (P < 0.05). Calcein fluorescence and histology revealed that greater mineral densities and better bone responses were achieved for groups B and C than for group A, with group C providing the best response. In conclusion, our Mg-Zn-Ca-alloy scaffold effectively aided bone repair. The group C scaffold exhibited the best corrosion resistance and osteogenesis properties, making it a candidate scaffold for repair of bone defects.

A study to compare the efficacy of polyether ether ketone rod device with titanium devices in posterior spinal fusion in a canine model.

BACKGROUND: The benefits of posterior lumbar fusion surgery with orthotopic paraspinal muscle-pediculated bone flaps are well established. However, the problem of non-union due to mechanical support is not completely resolved. The aim of the study was to compare the efficacy of polyether ether ketone (PEEK) rod device with conventional titanium devices in the posterior lumbar fusion surgery with orthotopic paraspinal muscle-pediculated bone flaps. METHODS: This was a randomized controlled study with an experimental animal model. Thirty-two mongrel dogs were randomly divided into two groups-control group (n = 16), which received the titanium device and the treatment group (n = 16), which received PEEK rods. The animals were sacrificed 8 or 16 weeks after surgery. Lumbar spines of dogs in both groups were removed, harvested, and assessed for radiographic, biomechanical, and histological changes. RESULTS: Results in the current study indicated that there was no significant difference in the lumbar spine of the control and treatment groups in terms of radiographic, manual palpation, and gross examination. However, certain parameters of biomechanical testing showed significant differences (p < 0.05) in stiffness and displacement, revealing a better fusion (treatment group showed decreased stiffness with decreased displacement) of the bone graft. Similarly, the histological analysis also revealed a significant fusion mass in both treatment and control groups (p < 0.05). CONCLUSIONS: These findings revealed that fixation using PEEK connecting rod could improve the union of the bone graft in the posterior lumbar spine fusion surgery compared with that of the titanium rod fixation.

Autogenous bone particle/titanium fiber composites for bone regeneration in a rabbit radius critical-size defect model.

PURPOSE: To explore the effects of autogenous bone particle/titanium fiber composites on repairing segmental bone defects in rabbits. MATERIALS AND METHODS: A model of bilateral radial bone defect was established in 36 New Zealand white rabbits which were randomly divided into 3 groups according to filling materials used for bilaterally defect treatment: in group C, 9 animal bone defect areas were prepared into simple bilateral radius bone defect (empty sham) as the control group; 27 rabbits were used in groups ABP and ABP-Ti. In group ABP, left defects were simply implanted with autogenous bone particles; meanwhile, group ABP-Ti animals had right defects implanted with autogenous bone particle/titanium fiber composites. Animals were sacrificed at 4, 8, and 12 weeks, respectively, after operation. RESULTS: Micro-CT showed that group C could not complete bone regeneration. Bone volume to tissue volume values in group ABP-Ti were better than group ABP. From histology and histomorphometry Groups ABP and ABP-Ti achieved bone repair, the bone formation of group ABP-Ti was better. The mechanical strength of group ABP-Ti was superior to that of other groups. CONCLUSIONS: These results confirmed the effectiveness of autologous bone particle/titanium fiber composites for promoting bone regeneration and mechanical strength.

Degraded and osteogenic properties of coated magnesium alloy AZ31; an experimental study.

BACKGROUND: Degraded and osteogenic property of coated magnesium alloy was evaluated for the fracture fixation in rabbits. METHODS: Magnesium alloy AZ31 with a different coating thickness by microarc oxidation was used, and the bilateral radial fracture model was created by the bite bone clamp. Thirty-six New Zealand white rabbits in weight of 2.5~3.0 kg were randomly divided into A, B, and C groups at four time points and other 3 rabbits as the control group without magnesium alloy. Coated magnesium alloy AZ31 was implanted on the fracture and fixed with silk thread. Indexes such as general observation, histology, X-ray, hematology, and mechanical properties were observed and detected at 2nd, 4th, 8th, and 12th week after implantation. RESULTS: Fracture in each rabbit was healed at 12th week after implantation. Among the three groups, the best results of general observation, histology, and X-ray appeared in A group without coating. However, A group showed the worst results from the perspective of mechanical properties about tensile strength and flexural strength, which failed to reach that of the natural bone at the 12th week. Comprehensive results displayed that C group with 20-µm coating was better than others in mechanical properties, while there is no difference between B and C groups in hematology. CONCLUSIONS: Degradation rate is inversely proportional to the coating thickness. And magnesium alloy with a 20-µm coating is more suitable for the fracture fixation.

BMP7 enhances the effect of BMSCs on extracellular matrix remodeling in a rabbit model of intervertebral disc degeneration.

Intervertebral discs (IVDs) provide stability and flexibility to the spinal column; however, IVDs, and in particular the nucleus pulposus (NP), undergo a degenerative process characterized by changes in the disc extracellular matrix (ECM), decreased cell viability, and reduced synthesis of proteoglycan and type II collagen. Here, we investigated the efficacy and feasibility of stem cell therapy using bone marrow mesenchymal stem cells (BMSCs) over-expressing bone morphogenetic protein 7 (BMP7) to promote ECM remodeling of degenerated IVDs. Lentivirus-mediated BMP7 over-expression induced differentiation of BMSCs into an NP phenotype, as indicated by expression of the NP markers collagen type II, aggrecan, SOX9 and keratins 8 and 19, increased the content of glycosaminoglycan, and up-regulated ß-1,3-glucuronosyl transferase 1, a regulator of chondroitin sulfate synthesis in NP cells. These effects were suppressed by Smad1 silencing, indicating that the effect of BMP7 on ECM remodeling was mediated by the Smad pathway. In vivo analysis in a rabbit model of disc degeneration showed that implantation of BMSCs over-expressing BMP7 promoted cell differentiation and proliferation in the NP, as well as their own survival, and these effects were mediated by the Smad pathway. The results of the present study indicate the beneficial effects of BMP7 on restoring ECM homeostasis in NP cells, and suggest potential strategies for improving cell therapy for the treatment of disc diseases.

Aquaporin 3 protects against lumbar intervertebral disc degeneration via the Wnt/ß-catenin pathway.

Previous studies have demonstrated that the expression of aquaporin 3 (AQP3), a water channel which promotes glycerol permeability and water transport across cell membranes, is reduced in degenerative lumbar intervertebral disc (IVD) tissues. However, the role of AQP3 in the pathogenesis of IVD degeneration has not recieved much scholarly attention. The objective of the present study was to investigate the effect of AQP3 on cell proliferation and extracellular matrix (ECM) degradation in human nucleus pulposus cells (hNPCs) using gain-of-function and loss-of-function experiments, and to determine whether Wnt/ß-catenin signaling is involved in the effect of AQP3 on IVD degeneration. hNPCs were transfected with the AQP3-pcDNA3.1 plasmid or AQP3 siRNA to overexpress or suppress AQP3. An MTT assay was performed to determine cell proliferation, and we found that AQP3 promoted hNPC proliferation. The expression of aggrecan, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)4 and ADAMTS5 was detected using western blot analysis, to examine the effect of AQP3 on ECM degradation in hNPCs. The results revealed that AQP3 inhibited ECM degradation in hNPCs. In addition, we found that Wnt/ß-catenin signaling was suppressed by AQP3. However, the effect of AQP3 on hNPC proliferation and ECM degradation was reversed by treatment with lithium chloride, a known activator of Wnt/ß­catenin signaling. In conclusion, using in vitro and in vivo tests, we have reported for the first time, to the best of our knowledge, that AQP3 exerts protective effects against IVD degeneration, and these are effected, at least partially, through the inhibition of Wnt/ß-catenin signaling.

Development and performance analysis of Si-CaP/fine particulate bone powder combined grafts for bone regeneration.

BACKGROUND: Although autogenous bone grafts as well as several bone graft substitute material have been used for some time, there is high demand for more efficient and less costly bone-substitute materials. Silicon-substituted calcium phosphates (Si-CaP) and fine particulate bone powder (FPBP) preparations have been previously shown to individually possess many of the required features of a bone graft substitute scaffold. However, when applied individually, these two materials fall short of an ideal substitute material. We investigated a new concept of combining Si-CaP with FPBP for improved performance in bone-repair. METHODS: We assessed Si-CaP/FPBP combined grafts in vitro, by measuring changes in pH, weight loss, water absorption and compressive strength over time. RESULTS: Si-CaP/FPBP combined grafts was found to produce conditions of alkaline pH levels compared to FPBP, and scaffold surface morphology conducive to bone cell adhesion, proliferation, differentiation, tissue growth and transport of nutrients, while maintaining elasticity and mechanical strength and degradation at a rate closer to osteogenesis. CONCLUSION: Si-CaP/FPBP combined grafts was found to be superior to any of the two components individually.

The fate of donor osteocytes in fine particulate bone powders during repair of bone defects in experimental rats.

The aim of the study was to investigate the fate of donor osteocytes in fine particulate bone powders during repair of bone defects in experimental rats. The iliac bone of male inbred DA rats was harvested and used as the larger bone grafts and also prepared as fine particulate (granulated) bone powders (300-500µm size particles) for transplantation into radial defects in female rats. The presence and relative amounts of genes specific to the sex-determining region of the Y-chromosome (Sry) originating from the bone grafts were evaluated by polymerase chain reaction and by in situ hybridization, respectively. Additional samples were evaluated histologically. In the larger bone grafts, the expression of Sry decreased relatively early, disappeared by 1 week, reappeared at 4 weeks and continued to increase with time. In the fine particulate bone powders, Sry was detected all the time and its expression was statistically greater than in the larger bone grafts at each time point. Both bone grafts provided donor cells to repair the defects. The donor cells seemed to function differently between the two groups. The fine particulate bone powders contained more living osteocytes in comparison with the larger bone grafts and may accelerate the healing of bone defects compared with conventional autografts.