Morphology Of Vascular Network Research Articles

Locational memory of macrovessel vascular cells is transcriptionally imprinted

Vascular pathologies show locational predisposition throughout the body; further insights into the transcriptomics basis of this vascular heterogeneity are needed. We analyzed transcriptomes from cultured endothelial cells and vascular smooth muscle cells from nine adult canine macrovessels: the aorta, coronary artery, vena cava, portal vein, femoral artery, femoral vein, saphenous vein, pulmonary vein, and pulmonary artery. We observed that organ-specific expression patterns persist in vitro, indicating that these genes are not regulated by blood flow or surrounding cell types but are likely fixed in the epigenetic memory. We further demonstrated the preserved location-specific expression of GATA4 protein in cultured cells and in the primary adult vessel. On a functional level, arterial and venous endothelial cells differed in vascular network morphology as the arterial networks maintained a higher complexity. Our findings prompt the rethinking of the extrapolation of results from single-origin endothelial cell systems.

Statistical and topological summaries aid disease detection for segmented retinal vascular images.

Disease complications can alter vascular network morphology and disrupt tissue functioning. Microvascular diseases of the retina are assessed by visual inspection of retinal images, but this can be challenging when diseases exhibit silent symptoms or patients cannot attend in-person meetings. We examine the performance of machine learning algorithms in detecting microvascular disease when trained on statistical and topological summaries of segmented retinal vascular images. We compute 13 separate descriptor vectors (5 statistical, 8 topological) to summarize the morphology of retinal vessel segmentation images and train support vector machines to predict each image's disease classification from the summary vectors. We assess the performance of each descriptor vector, using five-fold cross validation to estimate their accuracy. We apply these methods to four datasets that were assembled from four existing data repositories; three datasets contain segmented retinal vascular images from one of the repositories, whereas the fourth "All" dataset combines images from four repositories. Among the 13 total descriptor vectors considered, either a statistical Box-counting descriptor vector or a topological Flooding descriptor vector achieves the highest accuracy levels. On the combined "All" dataset, the Box-counting vector outperforms all other descriptors, including the topological Flooding vector which is sensitive to differences in the annotation styles between the different datasets. Our work represents a first step to establishing which computational methods are most suitable for identifying microvascular disease and assessing their current limitations. These methods could be incorporated into automated disease assessment tools.

Three-dimensional analyses of vascular network morphology in a murine lymph node by X-ray phase-contrast tomography with a 2D Talbot array.

With growing molecular evidence for correlations between spatial arrangement of blood vasculature and fundamental immunological functions, carried out in distinct compartments of the subdivided lymph node, there is an urgent need for three-dimensional models that can link these aspects. We reconstructed such models at a 1.84 µm resolution by the means of X-ray phase-contrast imaging with a 2D Talbot array in a short time without any staining. In addition reconstructions are verified in immunohistochemistry staining as well as in ultrastructural analyses. While conventional illustrations of mammalian lymph nodes depict the hilus as a definite point of blood and lymphatic vessel entry and exit, our method revealed that multiple branches enter and emerge from an area that extends up to one third of the organ's surface. This could be a prerequisite for the drastic and location-dependent remodeling of vascularization, which is necessary for lymph node expansion during inflammation. Contrary to corrosion cast studies we identified B-cell follicles exhibiting a two times denser capillary network than the deep cortical units of the T-cell zone. In addition to our observation of high endothelial venules spatially surrounding the follicles, this suggests a direct connection between morphology and B-cell homing. Our findings will deepen the understanding of functional lymph node composition and lymphocyte migration on a fundamental basis.

Investigation into the diversity in the fractal dimensions of arterioles and venules in a microvascular network – A quantitative analysis

Fractal dimension is a robust fractal parameter for estimating the morphology of vascular networks. It reflects the property of vascular networks that may vary and thus, differentiate between individual networks and/or identify physiological and pathological conditions. As such, fractal dimension differs also between arteriolar and venular compartments, yet the underlying reason is so far unclear. In order to understand the mechanisms behind these differences, we quantitatively analyzed the impacts of vessel attributes on the fractal dimension. Fractal dimension and vessel attributes given by vessel density (VD), vessel length density (VL), and diameter index (DI=VD/VL) were analyzed in three microvascular networks of the rat mesentery, which were reconstructed from experimental data. The results show that differences in diameter between arterioles and venules are primarily responsible for arterio-venous differences in fractal dimension. Moreover, multiple linear regression analysis demonstrates that the sensitivity of the variation of fractal dimension to vessel length and diameter varies with the type of the vessels. While the change of vessel length contributes 57.8 ± 3.4% to the variation of arteriolar dimension, vessel diameter contributes 63.9 ± 4.8% to the variation of venular dimension. The present study provides an explanation for the different fractal dimension and dimension variation in arteriolar and venular compartments. It highlights the importance of estimating the fractal dimensions of arterioles and venules separately, which will enhance the ability of feature extraction by fractal analysis in physiological and clinical application.

Fractal characterization of retinal microvascular network morphology during diabetic retinopathy progression.

The study aimed to characterize morphological changes of the retinal microvascular network during the progression of diabetic retinopathy. Publicly available retinal images captured by a digital fundus camera from DIARETDB1 and STARE databases were used. The retinal microvessels were segmented using the automatic method, and vascular network morphology was analyzed by fractal parametrization such as box-counting dimension, lacunarity, and multifractals. The results of the analysis were affected by the ability of the segmentation method to include smaller vessels with more branching generations. In cases where the segmentation was more detailed and included a higher number of vessel branching generations, increased severity of diabetic retinopathy was associated with increased complexity of microvascular network as measured by box-counting and multifractal dimensions, and decreased gappiness of retinal microvascular network as measured by lacunarity parameter. This association was not observed if the segmentation method included only 3-4 vessel branching generations. Severe stages of diabetic retinopathy could be detected noninvasively by using high resolution fundus photography and automatic microvascular segmentation to the high number of branching generations, followed by fractal analysis parametrization. This approach could improve risk stratification for the development of microvascular complications, cardiovascular disease, and dementia in diabetes.

Characterizing Branching Vascular Network Morphology in Polypoidal Choroidal Vasculopathy by Optical Coherence Tomography Angiography

This study analyze the morphological characteristics of branching vascular networks (BVN) in polypoidal choroidal vasculopathy (PCV) using optical coherence tomography angiography (OCTA), and correlate imaging characteristics with clinical presentations. We presented a retrospective observational case series for fifty cases of PCV confirmed by indocyanine green angiography. Macular OCTA were done by the AngioVue. The PCV cases were classified by distinct morphologic patterns of BVN by two retina specialists and clinical features were analyzed. The sensitivity of polyp detection by OCTA was 86% after manual segmentation and that of BVN was 90%. Three distinct morphologic patterns of BVN were identified. (1) The “Trunk” pattern (47%) exhibited major vessel trunk with features including presence of drusens, thin choroid, and larger BVN area. (2) The “Glomeruli” pattern (33%) showed anastomotic vascular network without major trunk. (3) The “Stick” pattern (20%) had localized BVN and the thickest choroid. Subtypes 2 and 3 held higher recurrence rate. In conclusions, the precise visualization of BVN on OCTA supported that OCTA might be a noninvasive tool to study the morphology of BVN in PCV, which exhibits three different morphological types. Identifying the morphology of BVN has the potential to prognosticate outcomes in PCV patients.

Quantitative Label-Free Imaging of 3D Vascular Networks Self-Assembled in Synthetic Hydrogels.

Vascularization is an important strategy to overcome diffusion limits and enable the formation of complex, physiologically relevant engineered tissues and organoids. Self-assembly is a technique to generate in vitro vascular networks, but engineering the necessary network morphology and function remains challenging. Here, autofluorescence multiphoton microscopy (aMPM), a label-free imaging technique, is used to quantitatively evaluate in vitro vascular network morphology. Vascular networks are generated using human embryonic stem cell-derived endothelial cells and primary human pericytes encapsulated in synthetic poly(ethylene glycol)-based hydrogels. Two custom-built bioreactors are used to generate distinct fluid flow patterns during vascular network formation: recirculating flow or continuous flow. aMPM is used to image these 3D vascular networks without the need for fixation, labels, or dyes. Image processing and analysis algorithms are developed to extract quantitative morphological parameters from these label-free images. It is observed with aMPM that both bioreactors promote formation of vascular networks with lower network anisotropy compared to static conditions, and the continuous flow bioreactor induces more branch points compared to static conditions. Importantly, these results agree with trends observed with immunocytochemistry. These studies demonstrate that aMPM allows label-free monitoring of vascular network morphology to streamline optimization of growth conditions and provide quality control of engineered tissues.

The Increased Transforming Growth Factor-β Signaling Induced by Diabetes Protects Retinal Vessels

The roles of transforming growth factor (TGF)-β in extracellular matrix production and vascular remodeling, coupled with increased TGF-β expression and signaling in diabetes, suggest TGF-β as an important contributor to the microangiopathy of diabetic retinopathy and nephropathy. To investigate whether increased TGF-β signaling could be a therapeutic target for preventing retinopathy, we used a pharmacologic approach (SM16, a selective inhibitor of the type 1 TGF-β receptor activin receptor-like kinase 5, orally active) to inhibit the increased, but not the basal, Tgf-β signaling in retinal vessels of diabetic rats. At the level of vascular gene expression, 3.5 months' diabetes induced minimal changes. Diabetes + SM16 for 3 weeks caused widespread changes in gene expression poised to enhance vascular inflammation, thrombosis, leakage, and wall instability; these changes were not observed in control ratsgiven SM16. The synergy of diabetes and SM16 in altering gene expression was not observed in the lung. At the level of vascular network morphology, 7 months' diabetes induced no detectable changes. Diabetes + SM16 for 3 weeks caused instead distorted morphology and decreased density. Thus, indiabetes, retinal vessels become dependent on a small increase in TGF-β signaling via activin receptor-like kinase 5 to maintain early integrity. The increased TGF-β signaling may protect against rapid retinopathy progression and should not be a target of inhibitory interventions.

Morphogenesis of 3D vascular networks is regulated by tensile forces

Understanding the forces controlling vascular network properties and morphology can enhance in vitro tissue vascularization and graft integration prospects. This work assessed the effect of uniaxial cell-induced and externally applied tensile forces on the morphology of vascular networks formed within fibroblast and endothelial cell-embedded 3D polymeric constructs. Force intensity correlated with network quality, as verified by inhibition of force and of angiogenesis-related regulators. Tensile forces during vessel formation resulted in parallel vessel orientation under static stretching and diagonal orientation under cyclic stretching, supported by angiogenic factors secreted in response to each stretch protocol. Implantation of scaffolds bearing network orientations matching those of host abdominal muscle tissue improved graft integration and the mechanical properties of the implantation site, a critical factor in repair of defects in this area. This study demonstrates the regulatory role of forces in angiogenesis and their capacities in vessel structure manipulation, which can be exploited to improve scaffolds for tissue repair.

Interstitial Fluid Flow and Drug Delivery in Vascularized Tumors: A Computational Model

Interstitial fluid is a solution that bathes and surrounds the human cells and provides them with nutrients and a way of waste removal. It is generally believed that elevated tumor interstitial fluid pressure (IFP) is partly responsible for the poor penetration and distribution of therapeutic agents in solid tumors, but the complex interplay of extravasation, permeabilities, vascular heterogeneities and diffusive and convective drug transport remains poorly understood. Here we consider–with the help of a theoretical model–the tumor IFP, interstitial fluid flow (IFF) and its impact upon drug delivery within tumor depending on biophysical determinants such as vessel network morphology, permeabilities and diffusive vs. convective transport. We developed a vascular tumor growth model, including vessel co-option, regression, and angiogenesis, that we extend here by the interstitium (represented by a porous medium obeying Darcy's law) and sources (vessels) and sinks (lymphatics) for IFF. With it we compute the spatial variation of the IFP and IFF and determine its correlation with the vascular network morphology and physiological parameters like vessel wall permeability, tissue conductivity, distribution of lymphatics etc. We find that an increased vascular wall conductivity together with a reduction of lymph function leads to increased tumor IFP, but also that the latter does not necessarily imply a decreased extravasation rate: Generally the IF flow rate is positively correlated with the various conductivities in the system. The IFF field is then used to determine the drug distribution after an injection via a convection diffusion reaction equation for intra- and extracellular concentrations with parameters guided by experimental data for the drug Doxorubicin. We observe that the interplay of convective and diffusive drug transport can lead to quite unexpected effects in the presence of a heterogeneous, compartmentalized vasculature. Finally we discuss various strategies to increase drug exposure time of tumor cells.

122 Fluorescence Videomicroscopic Visualization of Organ Neovascularisation with Special Reference to Cerebral Angiogenesis

The recruitment of new blood vessels in living tissues is essential for the pathogenesis and prognosis of organ disorders.The present study is concerned with intravital observations of newly formed vascular network in mouse brain, using fluo-rescence video-imaging. This visualization technique is a useful tool to study morphology and physiology of the newly developed vessels. The information obtained from the video-image includes the morphology of vascular network, the time required to develop blood vessels, and the velocity of red blood cells flowing in blood vessels.