Osteogenic differentiation of human adipose-derived stem cells : comparison of two different inductive media

Human mesenchymal stem cells (MSCs) have the potential to differentiate into cells of connective tissue lineages, including bone, cartilage, fat, muscle and also neurons. In our study we have examined the phenotypic profile of human adipose tissue-derived stem cells (hASCs) and compared different osteogenic-inductive media to assess hASC differentiation. Cells were enzymatically isolated from adipose tissues derived by liposuction from several adult human donors, purified and then expanded in culture. We obtained an abundant yield of hASCs with a constant proliferative trend, a doubling time of about 68 h and a mild variable clonogenic capacity. At passage 4, hASCs expressed MSC-related cell surface antigens (CD13, CD105, CD54, CD90, CD44), and subsequently hASCs were induced to differentiate into the osteogenic lineage for at least 3 weeks of culture in two distinct media, OM1 and OM2, differing in dexamethasone and ascorbic acid concentrations. Osteogenic differentiation of OM1- and OM2-cultured cells was assessed by evaluating cell morphology, osteopontin expression, alkaline phosphatase activity and calcium deposition. OM2 medium showed a higher osteogenic potential than OM1, as assessed by increased levels of calcium deposition, alkaline phospatase activity and osteopontin expression in comparison with OM1-differentiated cells. We conclude that hASCs efficiently differentiate into osteogenic lineage, particularly when cultured in inductive medium supplemented with 10 nM dexamethasone and 150 microM ascorbic acid

Efeitos Da Clonidina E Da Rilmenidina Sobre Os Sistemas Cardiorrespiratório E Gastrointestinal De Equinos

Clonidine and rilmenidine are drugs used in human medicine as central acting antihypertensive agents due to their actions on the alpha2-adrenoceptor and I1 imidazoline receptors in the central nervous system. Currently, clonidine is also used as a pre-anesthetic medication and in spinal anesthesia, and rilmenidine, despite the lower selectivity for alpha2-adrenergic receptors than clonidine, has also shown antinociceptive potential in experimental pain models. In this study, six horses were submitted to four treatments: R1 group (0.014 mg/kg of rilmenidine); R2 group (0.021 mg/kg of rilmenidine); Clo group (0.002 mg/kg of clonidine), and a control group. The assessment comprehended their heart and respiratory rates, systolic blood pressure and intestinal motility at basal levels and, then, 60 and 120 minutes after the oral administration of the drugs. Rilmenidine decreased heart rate on the two tested doses, which did not occur in the Clonidine treatment; slight variations in systolic blood pressure in all treatments and respiratory rate reduction in treatments with rilmenidine at 0.021 mg/ kg and clonidine at 0.002 mg/kg were also observed. Further studies with different dosages and varied administration routes are still necessary in order to elucidate more effects of these drugs on horses. © 2016, Cienc. anim. bras., Goiânia. All rights reserved.17460861

Human adipose-derived stem cells as future tools in tissue regeneration : osteogenic differentiation and cell-scaffold interaction

Tissue engineering is now contributing to new developments in several clinical fields, and mesenchymal stem cells derived from adipose tissue (hASCs) may provide a novel opportunity to replace, repair and promote the regeneration of diseased or damaged musculoskeletal tissue. Our interest was to characterize and differentiate hASCs isolated from twenty-three donors. Proliferation, CFU-F, cytofluorimetric and histochemistry analyses were performed. HASCs differentiate into osteogenic, chondrogenic, and adipogenic lineages, as assessed by tissue-specific markers such as alkaline phosphatase, osteopontin expression and deposition of calcium matrix, lipid-vacuoles formation and Glycosaminoglycans production. We also compared osteo-differentiated hASCs cultured on monolayer and loaded on biomaterials routinely used in the clinic, such as hydroxyapatite, cancellous human bone fragments, deproteinized bovine bone granules, and titanium. Scaffolds loaded with pre-differentiated hASCs do not affect cell proliferation and no cellular toxicity was observed. HASCs tightly adhere to scaffolds and differentiated-hASCs on human bone fragments and bovine bone granules produced, respectively, 3.4- and 2.1-fold more calcified matrix than osteo-differentiated hASCs on monolayer. Moreover, both human and deproteinized bovine bone is able to induce osteogenic differentiation of CTRL-hASCs. Although our in vitro results need to be confirmed in in vivo bone regeneration models, our data suggest that hASCs may be considered suitable biological tools for the screening of innovative scaffolds that would be useful in tissue engineering

Rapid Remission of Graves' Hyperthyroidism Without Thionamides Under Immunosuppressive Treatment for Concomitant Autoimmune Hepatitis.

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Bulk ion heating with ICRH in JET DT plasmas

Reactor relevant ICRH scenarios have been assessed during DT experiments on the JET tokamak using H mode divertor discharges with ITER-like shapes and safety factors. Deuterium minority heating in tritium plasmas was demonstrated for the first time. For 9% deuterium, an ICRH power of 6 MW gave 1.66 MW of fusion power from reactions between suprathermal deuterons and thermal tritons. The Q value of the steady state discharge reached 0.22 for the length of the RF flat-top (2.7 s), corresponding to three plasma energy replacement times. The Doppler broadened neutron spectrum showed a deuteron energy of 125 keV, which was optimum for fusion and close to the critical energy. Thus, strong bulk ion heating was obtained at the same time as high fusion efficiency. Deuterium fractions around 20% produced the strongest ion heating together with a strong reduction of the suprathermal deuteron tail. The ELMs had low amplitude and high frequency and each ELM transported less plasma energy content than the 1% required by ITER. The energy confinement time, on the ITERH97-P scale, was 0.90, which is sufficient for ignition in ITER. 3He minority heating, in approximately 50:50 D:T plasmas with up to 10% 3He, also demonstrated strong bulk ion heating. Central ion temperatures up to 13 keV were achieved, together with central electron temperatures up to 12 keV. The normalized H mode confinement time was 0.95. Second harmonic tritium heating produced energetic tritons above the critical energy. This scheme heats the electrons in JET, unlike in ITER where the lower power density will allow mainly ion heating. The inverted scenario of tritium minority ICRH in a deuterium plasma was demonstrated as a successful heating method producing both suprathermal neutrons and bulk ion heating. Theoretical calculations of the DT reactivity mostly give excellent agreement with the measured reaction rates