Effects of human umbilical cord mesenchymal stem cells on co‐cultured ovarian carcinoma cells

Ovarian carcinoma is mainly treated by surgery aided by chemotherapy. If supplemented by stem cells treatment, its recurrence rate and mortality rate will be decreased. This is a new therapy. In this study, ovarian cancer cells were cultured together with umbilical cord mesenchymal stem cells (UCMSCs), and the interactions between them were observed. The results showed that the survival rates of UCMSCs increased to 83.8 ± 2.2% from 56.5 ± 5.5%, and the survival rates of ovarian cancer cells decreased to 16.2 ± 2.2% from 43.5 ± 5.5% with the progression of the cultural time from 24 to 96 hr. There was a significant difference between them (p < .05). It revealed that UCMSCs could inhibit the proliferation of ovarian cancer cells.     

Immunoregulatory effects of human amniotic mesenchymal stem cells and their exosomes on human peripheral blood mononuclear cells

Background: The immunomodulatory effects of mesenchymal stem cells (MSCs) and their exosomes have been receiving increasing attention. This study investigated the immunoregulatory effects of human amniotic mesenchymal stem cells (hAMSCs) and their exosomes on phytohemagglutinin (PHA)-induced peripheral blood mononuclear cells (PBMCs). Methods: The hAMSCs used in the experiment were identified by light microscopy and flow cytometry, and the differentiation ability of the cells was determined by Oil Red O and Alizarin Red staining. The expressions of transforming growth factor (TGF)-β, indoleamine 2,3-dioxygenase (IDO), cyclooxygenase-2 (COX-2), hepatocyte growth factor (HGF), and interleukin (IL)-6 were detected by quantitative real-time polymerase chain reaction and western blotting. PBMCs, hAMSCs, and their exosomes were collected for in vitro group culture. Then the immunoregulatory ability of hAMSCs and their exosomes were analyzed by flow cytometry and Enzyme-linked immunosorbent assay. Results: The hAMSCs and exosomes were successfully extracted from the human amniotic membrane. TGF-β, IDO, COX-2, HGF, and IL-6 were significantly expressed in hAMSCs. In vitro co-culture showed that hAMSCs promoted the proliferation of Th2 cells in PHA-induced PBMCs, while hAMSCs and exosomes inhibited the secretion of TNF-α in PHA-induced PBMCs, and promoted the secretion of IL-4 and IL-10, and hAMSCs had more significant effects than exosomes. Conclusions: hAMSCs or exosomes could exert immunoregulatory effects on PHA-induced PBMCs by affecting Th2 cell proliferation and cytokine secretion.     

Effects of TGF‐β1 on mineralization mediated by rat calvaria‐derived osteogenic cells

In this study, we have analyzed the viability and cell growth, as well as, the mineralization of extracellular matrix (ECM) by alizarin red and von Kossa staining of calvaria‐derived osteogenic cultures, treated with TGF‐β1 alone or associated with Dex comparing with acid ascorbic (AA) + β‐glicerophosphate (βGP) (positive mineralization control). The expression of the noncollagenous proteins bone sialoprotein (BSP), osteopontin (OPN) and fibronectin (FN) were evaluated by indirect immunofluorescence. In addition, the main ultrastructural morphological findings were assessed by transmission electron microscopy. Osteogenic cells were isolated of calvaria bone from newborn (2‐day‐old) Wistar rats were treated with TGF‐β1 alone or with dexamethasone for 7, 10, and 14 days. As positive mineralization control, the cells were supplemented only with AA+ βGP. As negative control, the cells were cultured with basal medium (α‐MEM + 10%FBS + 1%gentamicin). The treatment with TGF‐β1, even when combined with Dex, decreased the viability and cell growth when compared with the positive control. Osteoblastic cell cultures were positive to alizarin red and von Kossa stainings after AA + βGP and Dex alone treatments. Positive immunoreaction was found for BSP, OPN and FN in all studied treatments. Otherwise, when the cell cultures were supplemented with TGF‐β1 and TGF‐β1 + Dex, no mineralization was observed in any of the studied periods. These present findings suggest that TGF‐β1, in the studied in vitro doses, inhibits the proliferation and differentiation of osteoblastic cells by impairment of nodule formation.     

Bones Morphogenic Protein‐4 and retinoic acid combined treatment comparative analysis for in vitro differentiation potential of murine mesenchymal stem cells derived from bone marrow and adipose tissue into germ cells

Nowadays, infertility is no longer considered as an unsolvable disorder due to progresses in germ cells derived from stem lineage with diverse origins. Technical and ethical challenges push researchers to investigate various tissue sources to approach more efficient gametes. The purpose of the current study is to investigate the efficacy of a combined medium, retinoic acid (RA) together with Bone Morphogenic Protein‐4 (BMP4), on differentiation of Bone Marrow Mesenchymal Stem Cells (BMMSCs) and adipose‐derived mesenchymal stem cells (ADMSCs) into germ cells. Murine MSCs were obtained from both Bone Marrow (BM) and Adipose Tissue (AT) samples and were analyzed for surface markers to get further verification of their nature. BMMSCs and ADMSCs were induced into osteogenic and adipogenic lineage cells respectively, to examine their multipotency. They were finally differentiated into germ cells using media enriched with BMP4 for 4 days followed by addition of RA for 7 days (11 days in total). Analyzing of differentiation potential of BMMSCs‐ and ADMSCs were performed via Immunofluorescence, Flowcytometry and Real time‐PCR techniques for germ cell‐specific markers (Mvh, Dazl, Stra8 and Scp3). Mesenchymal surface markers (CD90 and CD44) were expressed on both BMMSCs and ADMSCs, while endothelial and hematopoietic cell markers (CD31 and CD45) had no expression. Finally, all germ‐specific markers were expressed in both BM and AT. Although germ cells differentiated from ADMSCs showed faster growth and proliferation as well as easy collection, they significantly expressed germ‐specific markers lower than BMMSCs. This suggests stronger differentiation potential of murine BMMSCs than ADMSCs.     

Effects of aerobic training,resistance training,or combined resistance‐aerobic training on the left ventricular myocardium in a rat model

This study follows the left ventricular (LV) hypertrophy in rats undergoing aerobic training alone (A), resistance training alone (R), or combined resistance and aerobic training (RA) (usually referred as concurrent training) program. A sedentary control group (C) was included. LV remodeling was evaluated using electron and light microscopy. The LV weight to body weight (LVW: BW) increased 11.4% in A group, 35% in the R group, and 18% in the RA group compared to the C group. The LV thickness increased 6% in the A group, 17% in the R group, and 10% in the RA group. The LV internal diameter increased 19% in the A group, 3% in the R group, and 8% in the RA group compared with the C group. The cross‐sectional area of cardiomyocyte increased by 1% with the A group, 27% with R group, and 12% with RA training. The capillary density increased by 5.4% with A training, 11.0% with R training, and 7.7% with RA training compared with the C group. The volume fraction of interstitial collagen increased by 0.4% with training A, increased by 2.8% with R training, and 0.9% with RA training. In conclusion, except for the LV internal diameter, which increased more in the A group, the cardiac parameters increased more in the R group than in the other groups and in RA group than in A group. Collagen density increased from 5.4 ± 0.8% in the C group to 5.8 ± 0.6% in the A group (n. s.) (P > 0.05), to 8.2 ± 0.7% in the R group (P < 0.05), and to 6.3 ± 0.4% in the RA group (P < 0.05). These results demonstrate a significant increase for collagen content in the LV with R and RA exercise, but the increase was higher with R training alone than with RA training. Microsc. Res. Tech. 77:727–734, 2014. © 2014 Wiley Periodicals, Inc.     

<i>In vitro</i> study of emodin-induced nephrotoxicity in human renal glomerular endothelial cells on a microfluidic chip

Emodin is an effective component of rhubarb with positive pharmacological effects on human health. However, it is also toxic to different cells or tissues to varying degrees. The effects of emodin on glomerular endothelial cells (GECs) remain to be tested, and the documented works were always performed in vitro and hardly reflect the real physiological situation. To study the effects of emodin on GECs in a biomimetic environment, we utilized a microfluidic chip to assess the physiological reaction of human renal glomerular endothelial cells to various concentrations of emodin in this work. The results showed that emodin caused cytotoxicity, impaired glomerular filtration barrier integrity to macromolecules, and increased barrier permeability in a dose-dependent manner. With the increase in emodin concentration, the concentration of the pro-inflammatory cytokine tumor necrosis factor-α, interleukin (IL)-6, transforming growth factor-β1, and monocyte chemoattractant protein (MCP-1) increased while the production of inflammatory cytokine IL-6 first increased and then decreased with the increase in emodin concentration. Our findings shed new light on emodin-induced nephrotoxicity and provide insights for the application of microfluidic chip devices to reveal drug-cell interactions.     

Autophagosomal glycogen‐degrading activity and its relationship to the general autophagic activity in newborn rat hepatocytes: The effects of parenteral glucose administration

The effects of the administration of parenteral glucose on the postnatal glycogen autophagic activity and its relationship to the general autophagic activity, were studied in newborn rat liver using electron microscopy and biochemical methods. Glucose abolished the normal postnatal hypoglycemia and preserved the hepatocytic hyaloplasmic glycogen to the levels of birth. It also inhibited the normal postnatal increase in the number and volume of autophagic vacuoles. Glucose especially decreased the rate of postnatal development of the glycogen‐containing autophagic vacuoles. This decrease was greater than that of the autophagic vacuoles in general. In the control animals at the age of 6 h, the total volume of the glycogen‐containing autophagic vacuoles accounted for 87% of the autophagic vacuoles in general, whereas in the glucose‐treated animals of the same age, for only 62%. The results of this and previous studies support the view that the general autophagic activity that develops in the immediate postnatal period in rat hepatocytes is mainly expressed as glycogen autophagic activity selectively inhibited by glucose. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

The effect of natural products combination on MCF-7 cells exceeds tamoxifen therapeutic dose effects <i>in vitro</i>

Cancer remains to be one of the most severe sicknesses globally. Cases have kept rising over the years. Breast cancer (BC), which is among the leading types of cancers and predominantly affects women, is the second leading cause of cancer mortality. Researchers have developed interventions over the years; however, the BC survival rate has not improved since the 1980s. This has created the need for novel drug interventions that would manage and treat BC more effectively. This study focused on using a combination of natural product extracts such as phytoestrogen (Ziziphus jujube) and Tannin nanoparticles (NP99) together, which we have referred to as (Z.NP99) and tamoxifen (Tam) as one of the leading BC drugs since the 70s, in treating BC. The effectiveness of Tam if used alone in the treatment and if combined with Z.NP99 was evaluated using MCF-7 cells in vitro. The findings showed that the combination treatment of Z.NP99 affected the proliferation and viability of MCF-7 cells more than Tam at a 10 μg/mL dose. Moreover, Z.NP99 with Tam stimulated the maximum reduction of MCF-7 proliferation and viability in a time-dependent manner. Furthermore, Tam and Z.NP99 augmented the DNA fragmentation percentage combined with the upregulation of the apoptotic genes. Additionally, the results showed that the apoptotic impact of Z.NP99 and Tam on MCF-7 cells may be intermediated by down-regulating some genes such as Claudin-1 followed by down-regulating mRNA expression of MMP-9, VEGF, and BCL-2 genes of treated cells. Combining Tam with Z.NP99 considerably enhanced the effectiveness of conventional therapy. As a result, this study suggested that the Z.NP99 was ideal for developing effective natural treatments that would improve BC outcomes.     

The osteogenic effects of swimming on bone mass,strength, and microarchitecture in rats with unloading‐induced bone loss

The effect of nonweight‐bearing exercise on osteoporotic bones remains controversial and inconclusive. The purpose of this study was to evaluate the effects of swimming on osteoporotic tibias of rats submitted to hindlimb suspension. Initially, 20 Wistar rats were used to confirm a significant bone loss following 21 days of unloading. Thirty rats were then divided into 3 groups and followed during 51 days: CON (nonsuspended rats), S + WB (suspended rats for 21 days and then released for regular weight‐bearing) and, S + Swim (suspended rats for 21 days and then released from suspension and submitted to swimming exercise). We observed that swimming exercise was effective at fully recovering the bone deterioration caused by suspension, with significant increments in BMD, bone strength and bone volume. On the other hand, regular weight‐bearing failed at fully restoring the bone loss induced by unloading. These results indicate that swimming exercise may be a potential tool to improve bone density, strength, and trabecular volume in tibias with bone loss induced by mechanical unloading in suspended rats. We conclude that this modality of activity could be beneficial in improving bone mass, strength, and architecture in osteoporotic individuals induced by disuse, such as bed rest or those exposed to microgravity, who may not be able to perform weight‐bearing exercises. Microsc. Res. Tech. 78:784–791, 2015. © 2015 Wiley Periodicals, Inc.     

The effects of low sulfated ash,phosphorus and sulfur oils on camshaft/tappet tribocouples with various diamond‐like‐carbon coated tappets in motored and fired engines

This work was conducted as part of an EU FP‐7 project ‘Tailoring of Tribological Interfaces for Clean and Energy‐Efficient Diesel and Gasoline Powertrains’. The primary goal of this work is to reduce fuel consumption in internal combustion engines by the use of bespoke surface coatings and lubricants. Earlier work within the project had defined a series of oils and coatings to be evaluated in the final stage of the project. The results reported in this paper have used the OM646LA engine to evaluate wear and a motored cylinder head manufactured from the same engine type to evaluate the friction performance of coating/oil combinations. The coatings were applied to the tappets in the valvetrain and tested against uncoated camshafts. It will be shown that diamond‐like‐carbons can offer frictional benefits over an all‐steel system and that the choice of lubricant can incrementally reduce friction further. It will also be shown that while some diamond‐like‐carbon coatings are durable enough to survive the fired engine test intact, others are completely removed; interestingly, in all cases, the wear on the counter‐surface, the cam lobes, was in the order of a magnitude lower than for the standard all‐steel system. Copyright © 2014 John Wiley & Sons, Ltd.     

Systematic investigation of pipe flows and installation effects using laser Doppler anemometry—Part II. The effect of disturbed flow profiles on turbine gas meters—a describing empirical model

For systematic investigations of installation effects and for finding efficient ways to minimise these effects, a research project was initiated at the PTB. It covers the design of an automated test facility using a laser Doppler anemometer, the measurement of velocity profiles downstream of several pipe configurations and flow conditioners, as well as the measurement of the change in the gas meter behaviour, namely the shift of the error curve due to the disturbed velocity profiles.

Part I of this paper (presented in this issue) describes the test facility for the investigation of installation effects and shows the relation between pipe configuration and disturbed flow profile for a wide variety of pipe configurations and flow conditioners.

The second part compares the error shift of turbine meters with the characteristic of disturbed flow profiles. For this, three flow field parameters are used to quantify the disturbances of the velocity profiles such as the swirl intensity, flatness and asymmetry of the profile. Considering this, an empirical model is presented to explain the error shift of a turbine meter as a function of these three flow field parameters. The model will be verified for three types of turbine meters and the results will be discussed.