Jiefei Shen

Dedifferentiated fat cells： an alternative source of adult multipotent cells from the adipose tissues

When adipose‐derived stem cells (ASCs) are retrieved from the stromal vascular portion of adipose tissue, a large amount of mature adipocytes are often discarded. However, by modified ceiling culture technique based on their buoyancy, mature adipocytes can be easily isolated from the adipose cell suspension and dedifferentiated into lipid‐free fibroblast‐like cells, named dedifferentiated fat (DFAT) cells. DFAT cells re‐establish active proliferation ability and undertake multipotent capacities. Compared with ASCs and other adult stem cells, DFAT cells showed unique advantages in their abundance, isolation and homogeneity. In this concise review, the establishment and culture methods of DFAT cells are introduced and the current profiles of their cellular nature are summarized. Under proper induction culture in vitro or environment in vivo, DFAT cells could demonstrate adipogenic, osteogenic, chondrogenic and myogenic potentials. In angiogenic conditions, DFAT cells could exhibit perivascular characteristics and elicit neovascularization. Our preliminary findings also suggested the pericyte phenotype underlying such cell lineage, which supported a novel interpretation about the common origin of mesenchymal stem cells and tissue‐specific stem cells within blood vessel walls. Current research on DFAT cells indicated that this alternative source of adult multipotent cells has great potential in tissue engineering and regenerative medicine.

Characterization of a low shrinkage dental composite containing bismethylene spiroorthocarbonate expanding monomer.

In this study, a novel dental composite based on the unsaturated bismethylene spiroorthocarbonate expanding monomer 3,9-dimethylene-1,3,5,7-tetraoxa-spiro[5,5]undecane (BMSOC) and bisphenol-S-bis(3-meth acrylate-2-hydroxypropyl)ether (BisS-GMA) was prepared. CQ (camphorquinone) of 1 wt % and DMAEMA (2-(dimethylamino)ethyl methacrylate) of 2 wt % were used in a photoinitiation system to initiate the copolymerization of the matrix resins. Distilled water contact angle measurements were performed for the wettability measurement. Degree of conversion, volumetric shrinkage, contraction stress and compressive strength were measured using Fourier Transformation Infrared-FTIR spectroscopy, the AccuVol and a universal testing machine, respectively. Within the limitations of this study, it can be concluded that the resin composites modified by bismethylene spiroorthocarbonate and BisS-GMA showed a low volumetric shrinkage at 1.25% and a higher contact angle. The lower contraction stress, higher degree of conversion and compressive strength of the novel dental composites were also observed.

The effect of various sandblasting conditions on surface changes of dental zirconia and shear bond strength between zirconia core and indirect composite resin

PURPOSE To measure the surface loss of dental restorative zirconia and the short-term bond strength between an indirect composite resin (ICR) and zirconia ceramic after various sandblasting processes. MATERIALS AND METHODS Three hundred zirconia bars were randomly divided into 25 groups according to the type of sandblasting performed with pressures of 0.1, 0.2, 0.4 and 0.6 MPa, sandblasting times of 7, 14 and 21 seconds, and alumina powder sizes of 50 and 110 µm. The control group did not receive sandblasting. The volume loss and height loss on zirconia surface after sandblasting and the shear bond strength (SBS) between the sandblasted zirconia and ICR after 24-h immersion were measured for each group using multivariate analysis of variance (ANOVA) and Least Significance Difference (LSD) test (α=.05). After sandblasting, the failure modes of the ICR/zirconia surfaces were observed using scanning electron microscopy. RESULTS The volume loss and height loss were increased with higher sandblasting pressure and longer sandblasting treatment, but they decreased with larger powder size. SBS was significantly increased by increasing the sandblasting time from 7 seconds to 14 seconds and from 14 seconds to 21 seconds, as well as increasing the size of alumina powder from 50 µm to 110 µm. SBS was significantly increased from 0.1 MPa to 0.2 MPa according to the size of alumina powder. However, the SBSs were not significantly different with the sandblasting pressure of 0.2, 0.4 and 0.6 MPa. The possibilities of the combination of both adhesive failure and cohesive failure within the ICR were higher with the increases in bonding strength. CONCLUSION Based on the findings of this study, sandblasting with alumina particles at 0.2 MPa, 21 seconds and the powder size of 110 µm is recommended for dental applications to improve the bonding between zirconia core and ICR.

Effects of moderate static magnetic fields on the voltage-gated sodium and calcium channel currents in trigeminal ganglion neurons

Abstract Aim: To study the effects of static magnetic fields (SMF) on the electrophysiological properties of voltage-gated sodium and calcium channels on trigeminal ganglion (TRG) neurons. Methods: Acutely dissociated TRG neurons of neonatal SD rats were exposed to 125-mT and 12.5-mT SMF in exposure devices and whole-cell patch-clamp recordings were carried out to observe the changes of voltage-gated sodium channels (VGSC) and calcium channels (VGCC) currents, while laser scanning confocal microscopy was used to detect intracellular free Ca2+ concentration in TRG neurons, respectively. Results: (1) No obvious change of current–voltage (I–V) relationship and the peak current densities of VGSC and VGCC currents were found when TRG neurons were exposed to 125-mT and 12.5-mT SMF. However, the activation threshold, inactivation threshold and velocity of the channel currents above were significantly altered by 125-mT and 12.5-mT SMF. (2) The fluctuation of intracellular free Ca2+ concentration within TRG neurons were slowed by 125-mT and 12.5-mT SMF. When SMF was removed, the Ca2+ concentration level showed partial recovery in the TRG neurons previously exposed by 125-mT SMF, while there was a full recovery found in 12.5-mT-SMF-exposed neurons. Conclusions: Moderate-intensity SMF could affect the electrophysiological characteristics of VGCS and VGCC by altering their activation and inactivation threshold and velocity. The fluctuations of intracellular free Ca2+ caused by SMF exposure were not permanent in TRG neurons.

The phenotype and tissue-specific nature of multipotent cells derived from human mature adipocytes.

Dedifferentiated fat (DFAT) cells derived from mature adipocytes have been considered to be a homogeneous group of multipotent cells, which present to be an alternative source of adult stem cells for regenerative medicine. However, many aspects of the cellular nature about DFAT cells remained unclarified. This study aimed to elucidate the basic characteristics of DFAT cells underlying their functions and differentiation potentials. By modified ceiling culture technique, DFAT cells were converted from human mature adipocytes from the human buccal fat pads. Flow cytometry analysis revealed that those derived cells were a homogeneous population of CD13(+) CD29(+) CD105(+) CD44(+) CD31(-) CD34(-) CD309(-) α-SMA(-) cells. DFAT cells in this study demonstrated tissue-specific differentiation properties with strong adipogenic but much weaker osteogenic capacity. Neither did they express endothelial markers under angiogenic induction.

Serotonin acts as a novel regulator of interleukin-6 secretion in osteocytes through the activation of the 5-HT2B receptor and the ERK1/2 signalling pathway

Interleukin-6 (IL-6) is a potent stimulator of osteoclastic bone resorption. Osteocyte secretion of IL-6 plays an important role in bone metabolism. Serotonin (5-HT) has recently been reported to regulate bone metabolism. The aim of this study was to evaluate the effect of serotonin on osteocyte expression of IL-6. The requirement for the 5-HT receptor(s) and the role of the extracellular signal-regulated kinase 1/2 (ERK1/2) in serotonin-induced IL-6 synthesis were examined. In this study, real-time PCR and ELISA were used to analyse IL-6 gene and protein expression in serotonin-stimulated MLO-Y4 cells. ERK1/2 pathway activation was determined by Western blot. We found that serotonin significantly activated the ERK1/2 pathway and induced IL-6 mRNA expression and protein synthesis in cultured MLO-Y4 cells. However, these effects were abolished by pre-treatment of MLO-Y4 cells with a 5-HT2B receptor antagonist, RS127445 or the ERK1/2 inhibitor, PD98059. Our results indicate that serotonin stimulates osteocyte secretion of IL-6 and that this effect is associated with activation of 5-HT2B receptor and the ERK1/2 pathway. These findings provide support for a role of serotonin in bone metabolism by indicating serotonin regulates bone remodelling by mediating an inflammatory cytokine.

Concomitant activation of the PI3K/Akt and ERK1/2 signalling is involved in cyclic compressive force-induced IL-6 secretion in MLO-Y4 cells.

IL‐6 has a dual role in bone remodelling. The ERK1/2 pathway partially upregulated IL‐6 secretion in osteocyte‐like MLO‐Y4 cells exposed to CCF. We have now investigated the possible role of phosphatidylinositol 3‐kinase (PI3K)/Akt signalling pathway in the CCF‐induced IL‐6 expression. MLO‐Y4 cells were treated with CCF 2,000 µstrain, 2 Hz, or 10, 30 min, 1, 3 and 6 h. IL‐6 expression, Akt and ERK1/2 and PI3K/Akt phosphorylation were determined by RT‐PCR, ELISA and Western blotting. Inhibition of PI3K/Akt with LY294002 or ERK1/2 with PD98059 significantly attenuated IL‐6 upregulation, and IL‐6 expression was abolished by inhibiting both pathways. Inhibition of one pathway downregulated the others phosphorylation level. In conclusion, concomitant activation of PI3K/Akt and ERK1/2 pathways mediated IL‐6 expression in MLO‐Y4 cells under CCF.

Efficacy and safety of articaine versus lidocaine for irreversible pulpitis treatment: A systematic review and meta-analysis of randomised controlled trials.

The aim was to assess the efficacy and safety of articaine compared with lidocaine for irreversible pulpitis (IP) treatment. Databases were explored electronically and relevant journals as well as the references of the included studies were hand-searched for randomised clinical trials comparing the efficacy and safety of articaine with lidocaine in treatment of IP. Twenty studies were included, of which eight had low risk of bias, 10 had moderate risk of bias and two had high risk of bias. In comparison with 2% lidocaine with 1:100,000 epinephrine, 4% articaine with 1:100,000 epinephrine showed a higher success rate in anaesthesia of IP at either person (risk ratio (RR) 1.15; 95% confidence intervals (CI) 1.10 1.20; P < 0.00001) or tooth unit (RR 1.10; 95% CI 1.10 1.19, P < 0.00001), lower VAS scores during injection phase (mean difference (MD) -0.67; 95% CI -1.26 -0.08, P = 0.02) and treatment phase (MD -3.35; 95% CI -3.78 -2.91, P < 0.00001), shorter onset time of pulpal anaesthesia (MD -0.94; 95% CI -1.13 -0.74, P < 0.00001) and lower percentage of patients undergoing adverse events (RR 0.17; 95% CI 0.03 0.92, P = 0.04). Given the efficacy and safety of the two solutions, 4% articaine with 1:100,000 epinephrine was superior to 2% lidocaine with 1:100,000 epinephrine in dental treatments in IP.

Chlorogenic acid alters the voltage-gated potassium channel currents of trigeminal ganglion neurons

Chlorogenic acid (5-caffeoylquinic acid, CGA) is a phenolic compound that is found ubiquitously in plants, fruits and vegetables and is formed via the esterification of caffeic acid and quinic acid. In addition to its notable biological functions against cardiovascular diseases, type-2 diabetes and inflammatory conditions, CGA was recently hypothesized to be an alternative for the treatment of neurological diseases such as Alzheimers disease and neuropathic pain disorders. However, its mechanism of action is unclear. Voltage-gated potassium channel (Kv) is a crucial factor in the electro-physiological processes of sensory neurons. Kv has also been identified as a potential therapeutic target for inflammation and neuropathic pain disorders. In this study, we analysed the effects of CGA on the two main subtypes of Kv in trigeminal ganglion neurons, namely, the IK,A and IK,V channels. Trigeminal ganglion (TRG) neurons were acutely disassociated from the rat TRG, and two different doses of CGA (0.2 and 1 mmol⋅L−1) were applied to the cells. Whole-cell patch-clamp recordings were performed to observe alterations in the activation and inactivation properties of the IK,A and IK,V channels. The results demonstrated that 0.2 mmol⋅L−1 CGA decreased the peak current density of IK,A. Both 0.2 mmol⋅L−1 and 1 mmol⋅L−1 CGA also caused a significant reduction in the activation and inactivation thresholds of IK,A and IK,V. CGA exhibited a strong effect on the activation and inactivation velocities of IK,A and IK,V. These findings provide novel evidence explaining the biological effects of CGA, especially regarding its neurological effects.

Corticosterone Regulates the Expression of Neuropeptide Y and Reelin in MLO-Y4 Cells

Osteocytes that have a dendritic appearance are widely believed to form a complex cellular network system and play crucial roles in mechanotransduction as a principal bone mechanosensor, which is the basis of their neuronallike biology, as previously reported. Neuropeptide Y (NPY) and reelin mRNA, which are brain-specific neurogenic markers, have been identified in osteocytes. However, changes in the production of NPY and reelin in response to specific biochemical stimulation are unknown. In this study, we investigated the in vitro effect of corticosterone, one of the endogenous glucocorticoids, on the expression of NPY and reelin in the MLO-Y4 osteocyte cell line. Cells were treated with corticosterone at different concentrations (10−9 M-10−5 M) for 1, 3, 6, 12 and 24 h. As revealed, corticosterone reduced the MLO-Y4 cell viability and proliferation in a dose- and time-dependent manner based on an MTT assay and a Vi-CELL analyzer. The cells were then incubated with corticosterone (10−6 μM), and the NPY and reelin expression levels were detected at 1, 3, 6, 12 and 24 h using real-time PCR and Western blot analysis. These results demonstrated that at the gene and the protein levels, corticosterone significantly upregulated the NPY and reelin expression in a time-dependent manner. The application of a glucocorticoid receptor antagonist, RU486, reversed the reduced cell viability and the increased expression of NPY and reelin that were caused by corticosterone. To the best of our knowledge, this is the first report to verify that corticosterone regulates the NPY and reelin expression in osteocytes.