Fibroblast growth factor-2, but not the adipose tissue-derived stromal cells secretome, inhibits TGF-beta 1-induced differentiation of human cardiac fibroblasts into myofibroblasts

Transforming growth factor-beta 1 (TGF-beta 1) is a potent inducer of fibroblast to myofibroblast differentiation and contributes to the pro-fibrotic microenvironment during cardiac remodeling. Fibroblast growth factor-2 (FGF-2) is a growth factor secreted by adipose tissue-derived stromal cells (ASC) which can antagonize TGF-beta 1 signaling. We hypothesized that TGF-beta 1-induced cardiac fibroblast to myofibroblast differentiation is abrogated by FGF-2 and ASC conditioned medium (ASC-CMed). Our experiments demonstrated that TGF-beta 1 treatment-induced cardiac fibroblast differentiation into myofibroblasts, as evidenced by the formation of contractile stress fibers rich in alpha SMA. FGF-2 blocked the differentiation, as evidenced by the reduction in gene (TAGLN, p <0.0001; ACTA2, p = 0.0056) and protein (alpha SMA, p = 0.0338) expression of mesenchymal markers and extracellular matrix components gene expression (COL1A1, p <0.0001; COL3A1, p = 0.0029). ASC-CMed did not block myofibroblast differentiation. The treatment with FGF-2 increased matrix metalloproteinases gene expression (MMP1, p <0.0001; MMP14, p = 0.0027) and decreased the expression of tissue inhibitor of metalloproteinase gene TIMP2 (p = 0.0023). ASC-CMed did not influence these genes. The proliferation of TGF-beta 1-induced human cardiac fibroblasts was restored by both FGF-2 (p = 0.0002) and ASC-CMed (p = 0.0121). The present study supports the anti-fibrotic effects of FGF-2 through the blockage of cardiac fibroblast differentiation into myofibroblasts. ASC-CMed, however, did not replicate the anti-fibrotic effects of FGF-2 in vitro

Adipose tissue-derived stromal cells' conditioned medium modulates endothelial-mesenchymal transition induced by IL-1β/TGF-β2 but does not restore endothelial function

OBJECTIVES: Endothelial cells undergo TGF-β-driven endothelial-mesenchymal transition (EndMT), representing up to 25% of cardiac myofibroblasts in ischaemic hearts. Previous research showed that conditioned medium of adipose tissue-derived stromal cells (ASC-CMed) blocks the activation of fibroblasts into fibrotic myofibroblasts. We tested the hypothesis that ASC-CMed abrogates EndMT and prevents the formation of adverse myofibroblasts. MATERIALS AND METHODS: Human umbilical vein endothelial cells (HUVEC) were treated with IL-1β and TGF-β2 to induce EndMT, and the influence of ASC-CMed was assessed. As controls, non-treated HUVEC or HUVEC treated only with IL-1β in the absence or presence of ASC-CMed were used. Gene expression of inflammatory, endothelial, mesenchymal and extracellular matrix markers, transcription factors and cell receptors was analysed by RT-qPCR. The protein expression of endothelial and mesenchymal markers was evaluated by immunofluorescence microscopy and immunoblotting. Endothelial cell function was measured by sprouting assay. RESULTS: IL-1β/TGF-β2 treatment induced EndMT, as evidenced by the change in HUVEC morphology and an increase in mesenchymal markers. ASC-CMed blocked the EndMT-related fibrotic processes, as observed by reduced expression of mesenchymal markers TAGLN (P = 0.0008) and CNN1 (P = 0.0573), as well as SM22α (P = 0.0501). The angiogenesis potential was impaired in HUVEC undergoing EndMT and could not be restored by ASC-CMed. CONCLUSIONS: We demonstrated that ASC-CMed reduces IL-1β/TGF-β2-induced EndMT as observed by the loss of mesenchymal markers. The present study supports the anti-fibrotic effects of ASC-CMed through the modulation of the EndMT process

The integrated angular bispectrum

We study the position-dependent power spectrum and the integrated bispectrum statistic for 2D cosmological fields on the sphere (integrated angular bispectrum). First, we derive a useful, $m$-independent, formula for the full-sky integrated angular bispectrum, based on the construction of azimuthally symmetric patches. We then implement a pipeline for integrated angular bispectrum estimation, including a mean-field correction to account for spurious isotropy-breaking effects in realistic conditions (e.g., inhomogenous noise, sky masking). Finally, we show examples of applications of this estimator to CMB analysis, both using simulations and actual Planck data. Such examples include $f_\mathrm{NL}$ estimation, analyses of non-Gaussianity from secondary anisotropies (ISW-lensing and ISW-tSZ-tSZ bispectra) and studies of non-Gaussian signatures from foreground contamination.Comment: 38 pages, 14 figure

Bioactive decellularized cardiac extracellular matrix-based hydrogel as a sustained-release platform for human adipose tissue-derived stromal cell-secreted factors

The administration of trophic factors (TFs) released by mesenchymal stromal cells (MSCs) as therapy for cardiovascular diseases requires a delivery vehicle capable of binding and releasing the TF in a sustained manner. We hypothesized that hydrogels derived from cardiac decellularized extracellular matrix (cardiac dECM) bind MSC secretome-derived TF and release these in a sustained fashion. Pig-derived ventricular tissue was decellularized, milled to powder, digested, and assembled as a hydrogel upon warming at 37 degrees C. The conditioned medium (CMed) of adipose tissue-derived stromal cells (ASC) was collected, concentrated, and incorporated into the hydrogel at 1x, 10x, and 100x the original concentration. The release of 11 ASC-secreted factors (angiopoietin-1, angiopoietin-2, fibroblast growth factor-1, hepatocyte growth factor, platelet-derived growth factor-AA, vascular endothelial growth factor, interleukin-1 beta, interleukin-6, interleukin-8, CCL2, and matrix metalloproteinase-1) from hydrogels was immune assessed. Bioactivity was determined by endothelial cell proliferation, function, and assessment of endothelial mesenchymal transition. We showed that dECM hydrogels could be loaded with human ASC-secreted TFs, which are released in a sustained manner for several days subsequently. Different trophic factors had different release kinetics, which correlates with the initial concentration of CMed in the hydrogel. We observed that the more concentrated was the hydrogel, the more inflammation-related cytokines, and the less pro-regenerative TFs were released. Finally, we showed that the factors secreted by the hydrogel are biologically active as these influence cell behavior. The use of dECM hydrogels as a platform to bind and release paracrine factors secreted by (mesenchymal) cells is a potential alternative in the context of cardiovascular regeneration

Quijote-png: Quasi-maximum likelihood estimation of primordial non-gaussianity in the nonlinear dark matter density field

Future large-scale structure surveys are expected to improve current bounds on primordial non-Gaussianity (PNG), with a significant impact on our understanding of early universe physics. The level of such improvements will however strongly depend on the extent to which late-time nonlinearities erase the PNG signal on small scales. In this work, we show how much primordial information remains in the bispectrum of the nonlinear dark matter density field by implementing a new, simulation-based methodology for joint estimation of PNG amplitudes ( fNL) and standard Lambda cold dark matter parameters. The estimator is based on optimally compressed statistics, which, for a given input density field, combine power spectrum and modal bispectrum measurements, and numerically evaluate their covariance and their response to changes in cosmological parameters. In this first analysis, we focus on the matter density field, and we train and validate the estimator using a large suite of N-body simulations (QUIJOTE-PNG), including different types of PNG (local, equilateral, orthogonal)

Quijote-PNG: Optimizing the summary statistics to measure Primordial non-Gaussianity

We apply a suite of different estimators to the Quijote-PNG halo catalogues to find the best approach to constrain Primordial non-Gaussianity (PNG) at non-linear cosmological scales, up to $k_{\rm max} = 0.5 \, h\,{\rm Mpc}^{-1}$. The set of summary statistics considered in our analysis includes the power spectrum, bispectrum, halo mass function, marked power spectrum, and marked modal bispectrum. Marked statistics are used here for the first time in the context of PNG study. We perform a Fisher analysis to estimate their cosmological information content, showing substantial improvements when marked observables are added to the analysis. Starting from these summaries, we train deep neural networks (NN) to perform likelihood-free inference of cosmological and PNG parameters. We assess the performance of different subsets of summary statistics; in the case of $f_\mathrm{NL}^\mathrm{equil}$, we find that a combination of the power spectrum and a suitable marked power spectrum outperforms the combination of power spectrum and bispectrum, the baseline statistics usually employed in PNG analysis. A minimal pipeline to analyse the statistics we identified can be implemented either with our ML algorithm or via more traditional estimators, if these are deemed more reliable.Comment: 13 pages, 10 figure

Molecular and Biomechanical Clues From Cardiac Tissue Decellularized Extracellular Matrix Drive Stromal Cell Plasticity

Decellularized-organ-derived extracellular matrix (dECM) has been used for many years in tissue engineering and regenerative medicine. The manufacturing of hydrogels from dECM allows to make use of the pro-regenerative properties of the ECM and, simultaneously, to shape the material in any necessary way. The objective of the present project was to investigate differences between cardiovascular tissues (left ventricle, mitral valve, and aorta) with respect to generating dECM hydrogels and their interaction with cells in 2D and 3D. The left ventricle, mitral valve, and aorta of porcine hearts were decellularized using a series of detergent treatments (SDS, Triton-X 100 and deoxycholate). Mass spectrometry-based proteomics yielded the ECM proteins composition of the dECM. The dECM was digested with pepsin and resuspended in PBS (pH 7.4). Upon warming to 37°C, the suspension turns into a gel. Hydrogel stiffness was determined for samples with a dECM concentration of 20 mg/mL. Adipose tissue-derived stromal cells (ASC) and a combination of ASC with human pulmonary microvascular endothelial cells (HPMVEC) were cultured, respectively, on and in hydrogels to analyze cellular plasticity in 2D and vascular network formation in 3D. Differentiation of ASC was induced with 10 ng/mL of TGF-β1 and SM22α used as differentiation marker. 3D vascular network formation was evaluated with confocal microscopy after immunofluorescent staining of PECAM-1. In dECM, the most abundant protein was collagen VI for the left ventricle and mitral valve and elastin for the aorta. The stiffness of the hydrogel derived from the aorta (6,998 ± 895 Pa) was significantly higher than those derived from the left ventricle (3,384 ± 698 Pa) and the mitral valve (3,233 ± 323 Pa) (One-way ANOVA, p = 0.0008). Aorta-derived dECM hydrogel drove non-induced (without TGF-β1) differentiation, while hydrogels derived from the left ventricle and mitral valve inhibited TGF-β1-induced differentiation. All hydrogels supported vascular network formation within 7 days of culture, but ventricular dECM hydrogel demonstrated more robust vascular networks, with thicker and longer vascular structures. All the three main cardiovascular tissues, myocardium, valves, and large arteries, could be used to fabricate hydrogels from dECM, and these showed an origin-dependent influence on ASC differentiation and vascular network formation

Ethical Issues in the Use of Animal Models for Tissue Engineering:Reflections on Legal Aspects, Moral Theory, Three Rs Strategies, and Harm-Benefit Analysis

Animal experimentation requires a solid and rational moral foundation. Objective and emphatic decision-making and protocol evaluation by researchers and ethics committees remain a difficult and sensitive matter. This article presents three perspectives that facilitate a consideration of the minimally acceptable standard for animal experiments, in particular, in tissue engineering (TE) and regenerative medicine. First, we review the boundaries provided by law and public opinion in America and Europe. Second, we review contemporary moral theory to introduce the Neo-Rawlsian contractarian theory to objectively evaluate the ethics of animal experiments. Third, we introduce the importance of available reduction, replacement, and refinement strategies, which should be accounted for in moral decision-making and protocol evaluation of animal experiments. The three perspectives are integrated into an algorithmic and graphic harm-benefit analysis tool based on the most relevant aspects of animal models in TE. We conclude with a consideration of future avenues to improve animal experiments

Quijote PNG: The information content of the halo power spectrum and bispectrum

We investigate how much can be learnt about four types of primordial non-Gaussianity (PNG) from small-scale measurements of the halo field. Using the QUIJOTE-PNG simulations, we quantify the information content accessible with measurements of the halo power spectrum monopole and quadrupole, the matter power spectrum, the halo-matter cross spectrum and the halo bispectrum monopole. This analysis is the first to include small, non-linear scales, up to $k_\mathrm{max}=0.5 \mathrm{h/Mpc}$, and to explore whether these scales can break degeneracies with cosmological and nuisance parameters making use of thousands of N-body simulations. We perform all the halo measurements in redshift space with a single sample comprised of all halos with mass $>3.2 \times 10^{13}~h^{-1}M_\odot$. For local PNG, measurements of the scale dependent bias effect from the power spectrum using sample variance cancellation provide significantly tighter constraints than measurements of the halo bispectrum. In this case measurements of the small scales add minimal additional constraining power. In contrast, the information on equilateral and orthogonal PNG is primarily accessible through the bispectrum. For these shapes, small scale measurements increase the constraining power of the halo bispectrum by up to $\times4$, though the addition of scales beyond $k\approx 0.3 \mathrm{h/Mpc}$ improves constraints largely through reducing degeneracies between PNG and the other parameters. These degeneracies are even more powerfully mitigated through combining power spectrum and bispectrum measurements. However even with combined measurements and small scale information, equilateral non-Gaussianity remains highly degenerate with $\sigma_8$ and our bias model.Comment: Updated to accepted versio