Evaluation of New Octacalcium Phosphate-Coated Xenograft in Rats Calvarial Defect Model on Bone Regeneration.

Bone graft material is essential for satisfactory and sufficient bone growth which leads to a successful implant procedure. It is classified into autogenous bone, allobone, xenobone and alloplastic materials. Among them, it has been reported that heterogeneous bone graft material has a porous microstructure that increases blood vessels and bone formation, and shows faster bone formation than other types of bone graft materials. We observed new bone tissue formation and bone remodeling using Ti-oss® (Chiyewon Co., Ltd., Guri, Korea), a heterologous bone graft material. Using a Sprague-Dawley rat calvarial defect model to evaluate the bone healing effect of biomaterials, the efficacy of the newly developed xenograft Ti-oss® and Bio-Oss® (Geistilch Pharma AG, Wolhusen, Switzerland). The experimental animals were sacrificed at 8 and 12 weeks after surgery for each group and the experimental site was extracted. The average new bone area for the Ti-oss® experimental group at 8 weeks was 17.6%. The remaining graft material was 22.7% for the experimental group. The average new bone area for the Ti-oss® group was 24.3% at 12 weeks. The remaining graft material was 22.8% for the experimental group. It can be evaluated that the new bone-forming ability of Ti-oss® with octacalcium phosphate (OCP) has the bone-forming ability corresponding to the conventional products.

Effect of Lactobacillus rhamnosus GR-1 Supernatant on Cytokine and Chemokine Output From Human Amnion Cells Treated With Lipoteichoic Acid and Lipopolysaccharide.

Preterm birth occurs in 9% to 13% of all human pregnancies and accounts for 80% of all neonatal morbidities and mortalities. Approximately 40% of all preterm births are idiopathic and about half are associated with infection and/or an activated inflammatory process. Further to studies showing anti-inflammatory effects of supernatant from the probiotic Lactobacillus rhamnosus GR-1 (GR-1), we tested its ability to modulate cytokine and chemokine production from amnion cells in response to stimulation by bacterial wall components, lipopolysaccharide (LPS), and lipoteichoic acid (LTA). Placentae were collected from women undergoing elective cesarean section at term. Amnion cells were cultured for 48 hours to confluence, serum starved for 12 hours, and then treated with GR-1 supernatant (1:20 dilution), followed after 12 hours by LPS (100 ng/mL) or LTA (10 ng/mL) for an additional 12 hours. Both LTA and LPS caused significant increases in the concentration of the pro-inflammatory cytokine, tumor necrosis factor α (TNF-α; 103.9 ± 67.5 pg/mL and 368.3 ± 65.7 pg/mL, respectively) in medium from cultured amnion cells compared to control (<4 pg/mL). There was no significant effect of GR-1 supernatant alone on TNF-α output, but there was significant reduction after LPS treatment. The basal output of the immunomodulatory cytokine, interleukin 6, was 613 ± 170 pg/mL and increased significantly after addition of GR-1 supernatant, LTA, LPS, and combinations of LTA/LPS with GR-1 supernatant. In conclusion, probiotic L rhamnosus GR-1 attenuates the effect of both LPS and LTA in stimulating the output of the pro-inflammatory cytokine TNF-α from mixed cultures of human amnion cells in keeping with previous findings in human trophoblast cells.

Is there a role for probiotics in the prevention of preterm birth?

Preterm birth (PTB) continues to be a global health challenge. An over-production of inflammatory cytokines and chemokines, as well as an altered maternal vaginal microbiome has been implicated in the pathogenesis of inflammation/infection-associated PTB. Lactobacillus represents the dominant species in the vagina of most healthy pregnant women. The depletion of Lactobacillus in women with bacterial vaginosis (BV) has been associated with an increased risk of PTB. It remains unknown at what point an aberrant vaginal microbiome composition specifically induces the cascade leading to PTB. The ability of oral or vaginal lactobacilli probiotics to reduce BV occurrence and/or dampen inflammation is being considered as a means to prevent PTB. Certain anti-inflammatory properties of lactobacilli suggest potential mechanisms. To date, clinical studies have not been powered with sufficiently high rates of PTB, but overall, there is merit in examining this promising area of clinical science.

Facile synthesis and enhanced ultraviolet emission of ZnO nanorods prepared by vapor-confined face-to-face annealing.

In this study, we report a novel regrowth method of sol-gel-prepared ZnO films using a vapor-confined face-to-face annealing (VC-FTFA) technique in which mica was inserted between two films, followed by annealing with the FTFA method. The ZnO nanorods are regrown when zinc acetate dihydrate and zinc chloride (ZnCl2) are used as the solvent, because these generate ZnCl2 vapor. The near-band-edge emission intensity of the ZnO nanorods was enhanced through the VC-FTFA method, increasing significantly by a factor of 56 compared to that of ZnO films annealed in open air at 700 °C. Our method may provide a route toward the facile fabrication of ZnO nanorods.

Probiotic Lactobacillus rhamnosus GR-1 supernatant prevents lipopolysaccharide-induced preterm birth and reduces inflammation in pregnant CD-1 mice.

OBJECTIVE: The objective of this study was to determine the effect of probiotic Lactobacillus rhamnosus GR-1 supernatant (GR-1 SN) on lipopolysaccharide-induced preterm birth (PTB) and outputs of cytokines, chemokines, and progesterone in pregnant CD-1 mice. STUDY DESIGN: We compared PTB rates after intrauterine injection of lipopolysaccharide with and without previous GR-1 SN treatment. Cytokines and chemokines in the maternal plasma, myometrium, placenta, and amniotic fluid were examined with multiplex assay, and circulating maternal progesterone was measured with enzyme-linked immunoassay. Statistical significance was assessed with 2-tailed 1-way analysis of variance or analysis of variance on ranks. Fetal sex ratios in mice that delivered preterm were compared with those that delivered at term after lipopolysaccharide and GR-1 SN treatments. RESULTS: GR-1 SN reduced lipopolysaccharide-induced PTB by 43%. GR-1 SN significantly decreased the lipopolysaccharide-induced production of interleukin (IL)-1ß, -6, and -12p40, tumor necrosis factor-α, CCL4, and CCL5 in maternal plasma; IL-6, -12p70, -17, and -13 and tumor necrosis factor-α in myometrium; IL-6, -12p70, and -17 in placenta; and IL-6, tumor necrosis factor-α, CCL3, and CCL4 in amniotic fluid. Maternal plasma progesterone was reduced significantly after lipopolysaccharide injection with and without GR-1 SN pretreatment. There was no difference in fetal sex ratios between mice that delivered preterm and those that did not after lipopolysaccharide and GR-1 SN treatments. CONCLUSION: The supernatant of probiotic L rhamnosus GR-1 attenuated lipopolysaccharide-induced inflammation and PTB in vivo. GR-1 SN may confer therapeutic benefits in the prevention of infection-associated PTB by controlling systemic and intrauterine inflammation.

Lipopolysaccharide-Induced Profiles of Cytokine, Chemokine, and Growth Factors Produced by Human Decidual Cells Are Altered by Lactobacillus rhamnosus GR-1 Supernatant.

The aim of this study was to assess the effects of bacterial lipopolysaccharide (LPS) and Lactobacillus rhamnosus GR-1 supernatant (GR-1SN) on secretion profiles of cytokines, chemokines, and growth factors from primary cultures of human decidual cells. Lipopolysaccharide significantly increased the output of proinflammatory cytokines (interleukin [IL]-1B, IL-2, IL-6, IL-12p70, IL-15, IL-17A, interferon gamma [IFN-γ], and tumor necrosis factor [TNF]); anti-inflammatory cytokines (IL-1RN, IL-4, IL-9, and IL-10); chemokines (IL-8, eotaxin, IFN-inducible protein 10 [IP-10], monocyte chemoattractant protein 1 [MCP-1], macrophage inflammatory protein-1α [MIP-1α], macrophage inflammatory protein-1ß [MIP-1ß], and regulated on activation normal T cell expressed and secreted [RANTES]); and growth factors (granulocyte colony-stimulating factor [CSF] 3, CSF-2, and vascular endothelial growth factor A [VEGFA]). Lactobacillus rhamnosus GR-1SN alone significantly increased CSF-3, MIP-1α MIP-1ß, and RANTES but decreased IL-15 and IP-10 output. The GR-1SN also significantly or partially reduced LPS-induced proinflammatory cytokines TNF, IFN-γ, IL-1ß, IL-2 IL-6, IL-12p70, IL-15, IL-17, and IP-10; partially reduced LPS-induced anti-inflammatory cytokines IL-1RN, IL-4 and IL-10, and LPS-induced VEGFA output but did not affect CSF-3, MIP-1α, MIP-1ß, MCP-1, IL-8, and IL-9. Our results demonstrate that GR-1SN attenuates the inflammatory responses to LPS by human decidual cells, suggesting its potential role in ameliorating intrauterine infection.

Optical parameters of boron-doped ZnO nanorods grown by low-temperature hydrothermal reaction.

Sol-gel spin-coating was used to deposit ZnO seed layers onto quartz substrates, and ZnO nanorods doped with various concentrations of B (i.e., BZO nanorods) ranging from 0 to 2.0 at% were hydrothermally grown on the ZnO seed layers. The effects of B doping on the absorption coefficient, optical band gap, Urbach energy, refractive index, extinction coefficient, single-oscillator energy, dispersion energy, average oscillator strength, average oscillator wavelength, dielectric constant, and optical conductivity of the hydrothermally grown BZO nanorods were investigated. The optical band gaps were 3.255, 3.243, 3.254, 3.258, and 3.228 eV for the nanorods grwon at 0, 0.5, 1.0, 1.5 and 2.0 at% B, respectively. B doping increased the Urbach energy from 40.7 to 65.1 meV for the nanorods grown at 0 and 2.0 at% B, respectively, and significantly affected the dispersion energy, the single-oscillator energy, the average oscillator wavelength, the average oscillator strength, the refractive index, and the optical conductivity of the hydrothermally grown BZO nanorods.

Photoluminescence properties of defect emissions in Al-doped ZnO nanorod array thin films.

The power- and temperature-dependent photoluminescence properties of Al-doped ZnO nanorod array thin films grown by the hydrothermal method were investigated. The intensities of both the near-band-edge emission (NBE) and deep-level emission (DLE) as well as the overall spectral line shape were strongly affected by the excitation power. At low excitation power, the blue emission was found to show the highest intensity among the different emission lights. A low-temperature photoluminescence analysis revealed the bound-exciton-related luminescence peak at 3.362 eV. The dependence of peak energy with the excitation power indicates that these DLE processes are generated by DAP transitions. The overall intensity of DLE was found to decrease as the temperature increases. With regard to the blue emission (around 2.52 eV), it showed a well-pronounced shoulder at 200 K. The activation energy for this blue emission was 51.93 meV, which corresponds to the thermal dissociation energy required for the donor-acceptor pair transitions.

Structural and optical properties of ZnO nanorods on Mg0.2Zn0.8O seed layers grown by hydrothermal method.

ZnO nanorods were grown on the Mg0.2Zn0.8O seed layers with different thickness by hydrothermal method. Scanning electron microscopy (SEM), X-ray diffraction (XRD), and photoluminescence (PL) were carried out to investigate the effects of the Mg0.2Zn0.8O seed layer thickness on the structural and the optical properties of the ZnO nanorods. The residual stress in the Mg0.2Zn0.8O seed layers was depended on the thickness while the texture coefficient of the Mg0.2Zn0.8O seed layers was not affected significantly. The smaller full width at half maximum (FWHM) of the ZnO (002) diffraction and near-band-edge emission (NBE) peak and the larger average grain size were observed from the ZnO nanorods grown on the Mg0.2Zn0.8O seed layers with 5 layers (thickness of 350 nm), which indicate the enhancement the structural and the optical properties of the ZnO nanorods.

Noggin attenuates cerulein-induced acute pancreatitis and impaired autophagy.

OBJECTIVES: To investigate the role of bone morphogenetic protein (BMP) signaling in acute pancreatitis (AP) by administration of noggin, an endogenous BMP antagonist, in a cerulein-induced AP model. METHODS: Acute pancreatitis was induced by 9 hourly intraperitoneal injections of cerulein (50 µg/kg). Control mice received phosphate-buffered saline injections. In a separate group, noggin (0.5 mg/kg) was given intraperitoneally at 1 hour before and 2, 4, and 6 hours after AP induction. The mice were euthanized at 1 hour after completion of AP induction. The blood samples and the pancreas were harvested for analysis. Isolated pancreatic acini from normal mice and AR42J cells were treated with BMP2 and cerulein. AR42J cells were also treated with noggin. Phosphorylation of Smad1/5/8 was measured. RESULTS: Bone morphogenetic protein signaling was up-regulated in AP mouse pancreas. Bone morphogenetic protein 2 and cerulein-induced phosphorylation of Smad1/5/8 in the acinar cells in vitro, which was blocked by noggin. Noggin administration in vivo attenuated AP induction, decreased vacuole formation in acinar cells, blocked LC3-II levels, and partially restored Beclin-1 and lysosomal-associated membrane protein 2 levels. CONCLUSIONS: Bone morphogenetic protein signaling seems to promote AP induction and autophagy, as suggested by our study showing that noggin ameliorates AP and partially restores autophagic homeostasis.

Low-temperature growth of multiple-stack high-density ZnO nanoflowers/nanorods on plastic substrates.

Reported here is the low-temperature growth of multiple-stack high-density ZnO nanoflower/nanorod structures on polyethylene naphthalate (PEN) substrates derived from the surface modification of ZnO seed layers using an atmospheric-pressure plasma jet (APPJ) treatment. The plasma treatment could provide several advantages to the growth of multiple-stack ZnO nanoflower/nanorod structures: (i) the surface wettability of the seed layers changes from hydrophobic to hydrophilic, resulting in higher surface energies for the growth of high-density ZnO nanoflowers, (ii) the nucleation sites increase due to the increased surface roughness caused by the plasma etching, and (iii) there is no thermal damage to the plastic substrate from the plasma treatment due to its low-temperature weakly ionized discharge. It was also confirmed that multiple stacks of ZnO nanoflowers were obtained without degradation of the crystal quality or modification to the crystal shape or phase. The ZnO nanoflower/nanorod structures grew by lengths up to 4 µm due to an increased surface roughness of 10% and surface energy 5.5 times that of the seed layers. As shown, the APPJ is a very good method to obtain high-density ZnO nanostructures on plastic substrates below 150 °C, as is critical for flexible electronics.

Magnesium content ratio effects of MgxZn1-xO seed layers on structural and optical properties of ZnO nanorods.

ZnO nanorods were grown on MgxZn1-xO seed layers with different content ratio ranging from 0 to 0.3 by hydrothermal method. Scanning electron microscopy (SEM), X-ray diffraction (XRD), and photoluminescence (PL) were carried out to investigate the effects of Mg content ratio for the MgxZn1-xO seed layers on the structural and optical properties of the ZnO nanorods. The surface morphology and structural properties of the MgxZn1-xO seed layers were changed by the Mg incorporation. However, the appearance, such as density, diameter, and shape, of the ZnO nanorods grown on the MgxZn1-xO seed layers was not changed significantly. The highest intensity ratio of the near-band-edge emission (NBE) to deep-level emission (DLE) and the narrowest full width at half maximum (FWHM) of the NBE peaks, indicating improvement in the crystallinity and luminescent properties of the ZnO crystals, were observed in the ZnO nanorods grown on the MgxZn1_xO seed layers with the content ratio of the 0.05.

Temperature dependence of indigo light emission from mesoporous ZnO/porous silicon nanocomposites.

Nanocomposites of mesoporous zinc oxide (ZnO) and porous silicon (PS) were prepared through a hydrothermal method. Room-temperature (RT) and temperature-dependent photoluminescence (PL) were performed to investigate the optical properties and temperature dependence of the indigo emission peak from the ZnO/PS nanocomposites. An indigo emission peak from the nanocomposites and a red emission peak from the PS were observed in the case of the mesoporous ZnO/PS nanocomposites. At 10 K, the nanocomposites exhibited four emission peaks at 3.108, 2.929, 2.730, and 2.248 eV, which correspond to the DX, AX, DX-1LO, and DX-2LO phonon replicas, respectively. With an increase in temperature from 10 to 275 K, the curves in the intensities of the emission peaks formed an inverted "S" shape while their energies remained nearly constant. At 300 K, however, only the AX emission peak was observed; the DX and LO phonon replicas disappeared. The intensities of the DX and AX emission peaks exhibited anomalous behaviors.

Role of parathyroid hormone-related protein in the pro-inflammatory and pro-fibrogenic response associated with acute pancreatitis.

Pancreatitis is a common and potentially lethal necro-inflammatory disease with both acute and chronic manifestations. Current evidence suggests that the accumulated damage incurred during repeated bouts of acute pancreatitis (AP) can lead to chronic disease, which is associated with an increased risk of pancreatic cancer. While parathyroid hormone-related protein (PTHrP) exerts multiple effects in normal physiology and disease states, its function in pancreatitis has not been previously addressed. Here we show that PTHrP levels are transiently elevated in a mouse model of cerulein-induced AP. Treatment with alcohol, a risk factor for both AP and chronic pancreatitis (CP), also increases PTHrP levels. These effects of cerulein and ethanol are evident in isolated primary acinar and stellate cells, as well as in the immortalized acinar and stellate cell lines AR42J and irPSCc3, respectively. Ethanol sensitizes acinar and stellate cells to the PTHrP-modulating effects of cerulein. Treatment of acinar cells with PTHrP (1-36) increases expression of the inflammatory mediators interleukin-6 (IL-6) and intracellular adhesion protein (ICAM-1), suggesting a potential autocrine loop. PTHrP also increases apoptosis in AR42J cells. Stellate cells mediate the fibrogenic response associated with pancreatitis; PTHrP (1-36) increases procollagen I and fibronectin mRNA levels in both primary and immortalized stellate cells. The effects of cerulein and ethanol on levels of IL-6 and procollagen I are suppressed by the PTH1R antagonist, PTHrP (7-34). Together these studies identify PTHrP as a potential mediator of the inflammatory and fibrogenic responses associated with alcoholic pancreatitis.

Apoptosis of lung carcinoma cells induced by a flexible optical fiber-based cold microplasma.

Atmospheric pressure plasmas have been used as a therapy for cancer. However, the fairly large size and rigidity of present plasma-delivery systems obstructs the precise treatment of tumors in harder-to-reach internal organs such as the lungs, pancreas, and duodenum. In order to improve the targeted delivery of plasmas a highly flexible microplasma jet device is fabricated using a hollow-core optical fiber with an inner diameter of either 15 µm, 55 µm, or 200 µm. Described herein, based on this device, are results on lung carcinoma therapy using a microplasma cancer endoscope. Despite the small inner diameter and the low gas flow rate, the generated plasma jets are shown to be sufficiently effective to induce apoptosis, but not necrosis, in both cultured mouse lung carcinoma and fibroblast cells. Further, the lung carcinoma cells were found to be more sensitive to plasma treatment than the fibroblast cells based on the overall plasma dose conditions. This work enables directed cancer therapies using on highly flexible and precise hollow optical fiber-based plasma device and offers enhancements to microplasma cancer endoscopy using an improved method of plasma targeting and delivery.

Single-cell-level microplasma cancer therapy.

A flexible microplasma endoscope based on a 15 µm hollow-core glass optical fiber is fabricated, and tumor cell apoptotic analysis supports its potential use in targeted cancer therapies. The optical-fiber microplasma jet reveals antitumor activity at a certain plasma dose in animal studies.

Lactobacillus rhamnosus GR-1 stimulates colony-stimulating factor 3 (granulocyte) (CSF3) output in placental trophoblast cells in a fetal sex-dependent manner.

Bacterial vaginosis is associated with a 1.4-fold increased risk of preterm birth. We have shown previously that Lactobacillus rhamnosus GR-1 supernatant up-regulates interleukin 10 and down-regulates tumor necrosis factor-alpha output in lipopolysaccharide (LPS)-treated human primary placenta cultures in a fetal sex-dependent manner. We hypothesize that lactobacilli also exert their anti-inflammatory effect by up-regulation of colony-stimulating factor 3 (granulocyte) (CSF3), which is secreted from both immune and placental trophoblast cells, and that this activity is dependent on the sex of the fetus. Placental trophoblast cells were isolated from term elective cesarean section placentae using a Percoll gradient and separated from CD45(+) cells using magnetic purification. Cells were treated with LPS in the presence or absence of pretreatments with L. rhamnosus GR-1 supernatant or chemical inhibitors of the intracellular signaling pathways. Phosphorylations of mitogen-activated protein kinase 14 (MAPK14, previously known as p38) and signal transducer and activator of transcription (STAT) 3 were measured by Western blot analysis, and levels of CSF3 were determined by ELISA. CSF3 output was increased only in the placental trophoblast cells of female fetuses treated with LPS, GR-1 supernatant, and a combination of both treatments. The GR-1 supernatant up-regulated the phosphorylation of STAT3 and MAPK14. CSF3 output was inhibited by both Janus kinases (JAK) and MAPK14 inhibitors. None of the treatments was able to increase CSF3 output in either the pure trophoblast or the CD45(+) cell preparations alone. These results suggest an underlying mechanism for the sex difference in incidence of preterm birth and provide potential evidence for a therapeutic benefit of lactobacilli in reducing the risk of preterm labor.

Lactobacillus rhamnosus GR-1-induced IL-10 production in human placental trophoblast cells involves activation of JAK/STAT and MAPK pathways.

Intrauterine infection/inflammation complicates 25% to 40% of preterm births (PTB). The human vagina is normally populated by Lactobacillus species, some of which upregulate interleukin 10 (IL-10) output in lipopolysaccharide (LPS)-treated human placental trophoblast cells. We hypothesize that a probiotic strain, L rhamnosus GR-1 exerts its anti-inflammatory effect through activation of the Janus Kinases/Signal Transducers and Activators of Transcription (JAK/STAT) and mitogen-activated protein kinase (MAPK) pathways. Placental trophoblasts from term healthy pregnancies were treated with LPS in the presence or absence of pretreatments with GR-1 supernatant and/or chemical inhibitors of the intracellular signaling pathways. Phosphorylation of STAT3 and p38 was measured by Western Blot analysis, and output of IL-10 was determined by enzyme-linked immunosorbent assay (ELISA). Phosphorylation of STAT-3 and p38 was upregulated by GR-1 supernatant alone or in combination with LPS, while IL-10 output was inhibited by both JAK and p38 inhibitors. These data provide an underlying intracellular mechanism for cytokine regulation in the human placenta by L rhamnosus GR-1 and potential prevention of infection/inflammation-mediated PTB.

15-µm-sized single-cellular-level and cell-manipulatable microplasma jet in cancer therapies.

The authors describe a proposed 15-µm-sized, single-cellular-level, and cell-manipulatable microplasma jet device with a microcapillary glass tip and its potential in the development of cancer treatment therapies. The electrical and optical properties of the plasma jets and preliminary apoptosis results of cultured murine tumor cells and non-tumor fibroblast cells treated with the plasma jets are presented. The generated plasma jet was stable and enabled the treatment of cultured cells in cell culture plates regardless of the small inner diameter and low gas flow rate. The microplasma jet was observed inducing apoptosis in cultured murine melanoma tumor cells in a dose-dependent manner. Furthermore, the percentage of apoptotic cells of murine melanoma tumor cells induced by this plasma device was approximately 2.5 times bigger than that of murine fibroblast cells as indicated by an Annex V apoptosis assay. The apoptosis in cultured murine tumor cells by the 15-µm-sized single-cellular-level and cell-manipulatable microplasma jet device was also observed using an in situ apoptosis assay. We report on a novel microplasma jet device with the advantages of single-cellular-level and single cell-manipulatable plasma treatment with precise and solid stimuli. This highly precise plasma medicine, which enables new directed cancer therapies can be combined with current cell manipulation and cell culturing technologies without much difficulty.

The multifaceted effects of granulocyte colony-stimulating factor in immunomodulation and potential roles in intestinal immune homeostasis.

The three colony-stimulating factors, granulocyte/macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), and granulocyte colony-stimulating factor (G-CSF), have been regarded as immunostimulators because of their role in granulocyte and myeloid hematopoiesis and immune function. However, unlike GM-CSF and M-CSF, G-CSF possesses immunosuppressive effects on other immune cells including monocytes/macrophages, dendritic cells, and T lymphocytes when exogenously administered. Given the immunomodulatory effects of exogenous G-CSF, endogenous G-CSF may also play an important role in maintaining local immune homeostasis in tissue in which it is highly and constitutively produced. This review highlights the potential role of G-CSF in immunomodulation and intestinal immune homeostasis.