To assess the growth trajectory of preterm small-for-gestational-age (SGA) neonates compared to preterm non-small-for-gestational age neonates in the neonatal intensive care unit and special care ...nursery.
We conducted a retrospective cohort study at a large tertiary hospital in Victoria, Australia, examining neonates ≤34 weeks' gestation admitted to the neonatal intensive care unit or special care nursery between 2013 and 2017. We categorized neonates according to their birth weight centile: <10th centile (small-for-gestational age) and ≥10th centile (non-small-for-gestational age). Growth trajectory was tracked based on serial weights obtained in the neonatal intensive care unit and special care nursery, using z-scores derived from Fenton preterm growth charts. Our primary outcome was the change in weight z-score from birth to discharge from neonatal intensive care unit or special care nursery.
Of the 910 babies included, 88 were small-for-gestational age and 822 were appropriate-for gestational age. Both groups had a reduction in their weight z-score; however, SGA babies had a significantly smaller reduction (-0.62 SD compared to −0.85 SD, p < .0001). Small-for-gestational-age neonates were four times more likely to experience an increase in their weight z-score across their admission compared to neonates who were not small-for-gestational age (OR 4.04, 95% CI 2.23-7.48, p < .0001). Small-for-gestational-age neonates had an increased median length of stay, increased incidence of necrotizing enterocolitis but a reduced incidence of intraventricular hemorrhage.
Preterm SGA babies experience a smaller reduction in their weight trajectory compared to their appropriately grown counterparts in the neonatal intensive care unit or special care nursery.
Abstract only Cardiovascular disease (CVD) is the number one killer in Ireland and the wider EU. A hallmark of the disease is the obstruction to blood flow due to the build-up of vascular smooth ...muscle (SMCs)-like cells within the vessel wall. While polymer-coated DES have significantly reduced the incident of in-stent restenosis, current DESs lack the fundamental capacity for (i) adjustment of the drug dose and release kinetics and the (ii) ability to replenish the stent with a new drug on depletion. This limitation can be overcome by a strategy combining magnetic targeting via a uniform field-induced magnetization effect and a biocompatible magnetic nanoparticle (MNP) formulation designed for efficient entrapment and delivery of specific drugs that target the resident vascular stem cell source of the SMC. Magnetic nanoparticles (MNP’s) containing magnetite (Fe3O4) were fabricated, polymer coated with poly (DL-lactide-co-glycolide) polyvinyl alcohol PLGA-PVA and loaded with a γ-secretase inhibitor (GSI) of Notch signalling, DAPT using an oil in water emulsification technique. The free GSI’s and GSI-loaded MNP’s were assessed for drug release, the efficacy at controlling mesenchymal stem cell (MSC) growth (proliferation and apoptosis) and inhibiting myogenic differentiation under magnetic and non-magnetic conditions. The DAPT-loaded MNPs had an average hydrodynamic diameter of 351 d.nm Up to 40% of drug was released from MNPs within 48 h rising to 65% after 1 week under magnetic conditions. The Notch ligand, Jagged1 increased Hey1 mRNA levels and promoted myogenic differentiation of MSCs in vitro by increasing SMC differentiation markers, myosin heavy chain 11 (Myh11) and calponin1 (CNN1) expression, respectively. This effect was significantly attenuated following treatment of cells with MNP’s loaded with DAPT when compared to unloaded MNP’s. Notch GSI -loaded magnetic nanoparticles are functional at targeting vascular stem cells in vitro.
The accumulation of vascular smooth muscle (SMC)-like cells and stem cell-derived myogenic and osteogenic progeny contributes significantly to arteriosclerotic disease. This study established whether ...label-free vibrational spectroscopy can discriminate de-differentiated ‘synthetic’ SMCs from undifferentiated stem cells and their myogenic and osteogenic progeny in vitro, compared with conventional immunocytochemical and genetic analyses. TGF-β1- and Jagged1-induced myogenic differentiation of CD44+ mesenchymal stem cells was confirmed in vitro by immunocytochemical analysis of specific SMC differentiation marker expression (α-actin, calponin and myosin heavy chain 11), an epigenetic histone mark (H3K4me2) at the myosin heavy chain 11 locus, promoter transactivation and mRNA transcript levels. Osteogenic differentiation was confirmed by alizarin red staining of calcium deposition. Fourier Transform Infrared (FTIR) maps facilitated initial screening and discrimination while Raman spectroscopy of individual cell nuclei revealed specific spectral signatures of each cell type in vitro, using Principal Components Analysis (PCA). PCA fed Linear Discriminant Analysis (LDA) enabled quantification of this discrimination and the sensitivity and specificity value was determined for all cell populations based on a leave-one-out cross validation method and revealed that de-differentiated SMCs and stem-cell derived myogenic progeny in culture shared the greatest similarity. FTIR and Raman spectroscopy discriminated undifferentiated stem cells from both their myogenic and osteogenic progeny. The ability to detect stem cell-derived myogenic progeny using label-free platforms in situ may facilitate interrogation of these important phenotypes during vascular disease progression.
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•TGF-β1 and the Notch ligand, Jagged1 both induced myogenic differentiation of CD44+ mesenchymal stem cells in vitro•FTIR maps facilitated initial screening while Raman spectroscopy of individual cell nuclei revealed specific spectral signatures using PCA•PCA fed LDA enabled quantification of discrimination of undifferentiated stem cells from both their myogenic and osteogenic progeny.•De-differentiated SMCs shared the greatest spectral similarity with stem-cell derived myogenic progeny following cross validation•Label-free discrimination in situ may facilitate interrogation of these important phenotypes during vascular disease progression
Abstract only Background: The source of intimal vascular smooth muscle cells (SMCs) following vascular remodelling has been controversial, with either de-differentiated SMCs and/or stem cell-derived ...SMCs playing a putative role. Fourier transform Infrared (FTIR) and Raman spectroscopy are complementary forms of vibrational spectroscopy which provide an excellent platform for extracting important biochemical data in a label-free manner to discriminate cell types. Aim: Determine whether native differentiated SMCs can be distinguished from mesenchymal stem cells (MSCs) and MSC-derived SMCs using vibrational spectroscopy. Methods: Freshly isolated rat aortic differentiated SMCs (up to passage 4), CD44+ bone marrow derived mesenchymal stem cells (MSCs), MSC-derived smooth muscle cells (mdSMCs - after TGF-β1 treatment for 14 d) and osteoblasts (mdOSTs - after osteogenic inductive stimulation for 21 d) were grown and fixed on calcium fluoride slides before their respective spectra were recorded by Raman and FTIR Spectroscopy. Multivariate statistical algorithms, including Principal Components Analysis (PCA) and Linear Discriminant Analysis (LDA), were applied to the spectra in order to classify the cell types based on their biochemical variation. Results: The recorded spectra for each cell type revealed significant visible differences between the cells across all recordings. The PCA score plot discriminated the cells based on their unique characteristics. A combination of PCA-LDA were applied for classification, and a leave one out cross validation resulted in sensitivities and specificities that were >95%. Conclusion: Vibrational spectroscopy discriminates differentiated SMCs from MSC and their vascular progeny and may be useful for identifying these cells as early biomarkers of disease.
Abstract only The accumulation of vascular smooth muscle (SMC)-like cells within the intima contributes significantly to intimal medial thickening (IMT) and vascular remodeling typical of ...arteriosclerotic disease. Light has emerged as a powerful tool to interrogate cells label-free and facilitates discriminant observations both in vitro and in vivo . The auto-fluorescence (AF) profile of individual cells isolated from arteriosclerotic vessels, captured on V-cup array and interrogated across five wavelengths using a novel Lab-on-a-Disc platform, was significantly increased at the 565 ± 20nm wavelength concomitant with a reduction in Myh11 expression, when compared to differentiated vascular smooth muscle (SMC) cells from control vessels. In vitro, TGF-β1 promoted myogenic differentiation of murine bone-marrow derived Sca1 + /CD44 + mesenchymal stem cells (MSC) and murine Sca1 + C3H 10T1/2 cells concomitant with enrichment of the specific SMC epigenetic histone mark, H3K4me2 at the Myh11 promoter, Myh11 promoter transactivation and increased SMC differentiation marker mRNA and protein expression. Myogenic differentiation resulted in a significant increase in the AF intensity across 565 ± 20nm wavelength, an effect not observed for TGF-β1 treated RAMOS human B lymphocytes but mimicked by Notch activation of resident Sca1 + multipotent vascular stem cells (MVSCs) with Jagged1 and inhibited following elastin and collagen III depletion, respectively. Moreover, the temporal increase in the AF intensity at 565 ± 20nm wavelength during myogenic differentiation was similar to the AF profile of dissociated cells from arteriosclerotic vessels at this same wavelength. These data suggest that an AF photonic fingerprint of stem cell-derived myogenic progeny in vitro mimics that of vascular cells ex vivo following injury.
A hallmark of subclinical atherosclerosis is the accumulation of vascular smooth muscle cell (SMC)-like cells leading to intimal thickening and lesion formation. While medial SMCs contribute to ...vascular lesions, the involvement of resident vascular stem cells (vSCs) remains unclear. We evaluated single cell photonics as a discriminator of cell phenotype in vitro before the presence of vSC within vascular lesions was assessed ex vivo using supervised machine learning and further validated using lineage tracing analysis
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Using a novel lab-on-a-Disk(Load) platform, label-free single cell photonic emissions from normal and injured vessels ex vivo were interrogated and compared to freshly isolated aortic SMCs, cultured Movas SMCs, macrophages, B-cells, S100β
+
mVSc, bone marrow derived mesenchymal stem cells (MSC) and their respective myogenic progeny across five broadband light wavelengths (λ465 - λ670 ± 20 nm). We found that profiles were of sufficient coverage, specificity, and quality to clearly distinguish medial SMCs from different vascular beds (carotid vs aorta), discriminate normal carotid medial SMCs from lesional SMC-like cells ex vivo following flow restriction
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and identify SMC differentiation of a series of multipotent stem cells following treatment with transforming growth factor beta 1 (TGF- β1), the Notch ligand Jagged1, and Sonic Hedgehog using multivariate analysis, in part, due to photonic emissions from enhanced collagen III and elastin expression. Supervised machine learning supported genetic lineage tracing analysis of S100β
+
vSCs and identified the presence of S100β
+
vSC-derived myogenic progeny within vascular lesions. We conclude disease-relevant photonic signatures may have predictive value for vascular disease.
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