Blood Capillary Network Research Articles

Fluorescent Pdots Facilitate High-Resolution Mapping of the Intact Meningeal Vascular Network and Eye-Brain Connections.

Meningeal vascular network is significant in neurology and neurosurgery. However, high-resolution imaging of intact meningeal vascular network is lacking. In this work, we develop a practical experimental method to ensure that the intact meninges are morphologically unfolded and fixed in an agarose gel. With the help of high-brightness polymer dots (Pdots) as probe, macroscopic and detailed imaging of the vascular network on the intact dorsal meninges can be performed. Meningeal vessels are symmetrically distributed along the superior sagittal sinus, and the distribution of meningeal vessels had a certain degree of hierarchy. The meninges are thicker blood vessels and capillary networks from the outside to the inside. Moreover, the diameter of the capillaries is 3.96 ± 0.89 μm. Interestingly, meningeal primo vessels in the central nervous system of mice is imaged with the diameter of 4.18 ± 1.18 μm, which has not been reported previously. It is worth mentioning that we found that orthotopic xenografts of brain tumors caused the appearance of corneal neovascularization and morphological changes in optic nerve microvessels. In conclusion, our work provides an effective Pdots-based imaging method for follow-up research on meningeal vascular-related diseases, and illustrates that the eye can serve as a window for the prevention and diagnosis of brain diseases.

On-Site Construction of a Full-Thickness Skin Equivalent with Endothelial Tube Networks via Multilayer Electrospinning for Wound Coverage.

Novel full-thickness skin substitutes are of increasing interest due to the inherent limitations of current models lacking capillary networks. Herein, we developed a novel full-thickness skin tissue containing blood capillary networks through a layer-by-layer assembly approach using a handy electrospinning apparatus and evaluated its skin wound coverage potential in vivo. The average diameter and thickness of fabricated poly-ε-caprolactone-cellulose acetate scaffolds were easily tuned in the range of 474 ± 77-758 ± 113 nm and 9.43 ± 2.23-29.96 ± 5.78 μm by varying electrospinning distance and duration, as indicated by FE-SEM. Besides, keratinocytes exhibited homogeneous differentiation throughout the fibrous matrix prepared with electrospinning distance and duration of 9 cm and 1.5 min within five-layer (5L) epidermal tissues with thickness of 135-150 μm. Moreover, coculture of vascular endothelial cells, circulating fibrocytes, and fibroblasts within the 5L dermis displayed network formation in vitro, resulting in reduced inflammatory factor levels and enhanced integration with the host vasculature in vivo. Additionally, the skin equivalent grafts consisting of the epidermal layer, biomimetic basement membrane, and vascularized dermis layer with an elastic modulus of approximately 11.82 MPa exhibited accelerated wound closure effect indicative of re-epithelialization and neovascularization with long-term cell survival into the host, which was confirmed by wound-healing rate, bioluminescence imaging activity, and histological analysis. It is the first report of a full-thickness skin equivalent constructed using a battery-operated electrospinning apparatus, highlighting its tremendous potential in regenerative medicine.

Control of blood capillary networks and holes in blood-brain barrier models by regulating elastic modulus of scaffolds.

The blood-brain barrier (BBB) is a type of capillary network characterized by a highly selective barrier, which restricts the transport of substances between the blood and nervous system. Numerous in vitro models of the BBB have been developed for drug testing, but a BBB model with controllable capillary structures remains a major challenge. In this study, we report for the first time a unique method of controlling the blood capillary networks and characteristic holes formation in a BBB model by varying the elastic modulus of a three-dimensional scaffold. The characteristic hole structures are formed by the migration of endothelial cells from the model surface to the interior, which have functions of connecting the model interior to the external environment. The hole depth increased, as the elastic modulus of the fibrin gel scaffold increased, and the internal capillary network length increased with decreasing elastic modulus. Besides, internal astrocytes and pericytes were also found to be important for inducing hole formation from the model surface. Furthermore, RNA sequencing indicated up-regulated genes related to matrix metalloproteinases and angiogenesis, suggesting a relationship between enzymatic degradation of the scaffolds and hole formation. The findings of this study introduce a new method of fabricating complex BBB models for drug assessment.

A mathematical characterization of anatomically consistent blood capillary networks

Blood microcirculation is the site of control of tissue perfusion, blood-tissue exchange, and tissue blood volume. Despite the many irregularities, almost ubiquitously, one can recognize in microcirculation vessels a hierarchy of arterioles and venules, organized in tree-like structures, and capillary plexi, organized in net-like structures. Whilst for arterioles and venules it may be envisageable to obtain geometries needed for numerical simulations from imaging techniques, the size and numerosity of capillaries makes this task much more cumbersome. For this reason, it is interesting to study approaches to generate in silico-derived artifacts of capillary networks, even in view of machine-learning based approaches which require a large amount of samples for training. Artificial networks must correctly reflect proper metrics and topology, which in turn, will ensure with proper boundary conditions a physiological blood flux in the net and a sufficient nutrient distribution in the surrounding tissues. In this paper, we introduce the sequence of curves whose limit is the space filling Hilbert curve and we discuss its inherent properties and we obtain the backbone of the artificial capillary network from a suitable element of this sequence. The backbone represents a significant synthesis of basic metric features of the network and, in this context, its properties can be studied analytically. In this framework, the Hilbert curve is a malleable entity which allows to shape the backbone according to the physical indicators. In particular, two significant factors are shown to control the network topology and scaling: the iteration step of the Hilbert curve generation and the characteristic length of the REV, respectively. Based on the points we generate for a certain iteration step, we then obtain via spline interpolation a smoothed version of the curve, which fine–tunes the tortuosity. A volumetric construction is obtained building a tubular neighborhood of the backbone, whose metrics can be computed and tuned as well. Numerical simulations of the blood flow in the obtained geometry show the physical fields occurring in the artificial network.

Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold

Due to a relative paucity of studies on human lymphatic assembly in vitro and subsequent in vivo transplantation, capillary formation and survival of primary human lymphatic (hLEC) and blood endothelial cells (hBEC) ± primary human vascular smooth muscle cells (hvSMC) were evaluated and compared in vitro and in vivo. hLEC ± hvSMC or hBEC ± hvSMC were seeded in a 3D porous scaffold in vitro, and capillary percent vascular volume (PVV) and vascular density (VD)/mm2 assessed. Scaffolds were also transplanted into a sub-cutaneous rat wound with morphology/morphometry assessment. Initially hBEC formed a larger vessel network in vitro than hLEC, with interconnected capillaries evident at 2 days. Interconnected lymphatic capillaries were slower (3 days) to assemble. hLEC capillaries demonstrated a significant overall increase in PVV (p = 0.0083) and VD (p = 0.0039) in vitro when co-cultured with hvSMC. A similar increase did not occur for hBEC + hvSMC in vitro, but hBEC + hvSMC in vivo significantly increased PVV (p = 0.0035) and VD (p = 0.0087). Morphology/morphometry established that hLEC vessels maintained distinct cell markers, and demonstrated significantly increased individual vessel and network size, and longer survival than hBEC capillaries in vivo, and established inosculation with rat lymphatics, with evidence of lymphatic function. The porous polyurethane scaffold provided advantages to capillary network formation due to its large (300–600 μm diameter) interconnected pores, and sufficient stability to ensure successful surgical transplantation in vivo. Given their successful survival and function in vivo within the porous scaffold, in vitro assembled hLEC networks using this method are potentially applicable to clinical scenarios requiring replacement of dysfunctional or absent lymphatic networks.

A Dynamic, In Vitro BBB Model to Study the Effects of Varying Levels of Shear Stress.

The blood-brain barrier (BBB) consists of a tight network of blood capillaries in the brain that separate the circulatory system from the central nervous system. Its particular properties are based on the dynamic interaction between cerebral endothelial cells and other surrounding cells, especially astrocytes. We have designed and synthesized a three-dimensional scaffold that recapitulates the main hallmarks of the BBB extracellular matrix and serves as a platform to co-culture human brain microvascular endothelial cells and human cortical astrocytes. The scaffold can be exposed to flow, thereby allowing the study of flow-mediated pathways at the BBB.

Detailed morphology of tentacular apparatus and central nervous system in Owenia borealis (Annelida, Oweniidae)

The Oweniidae are marine annelids with many unusual features of organ system, development, morphology, and ultrastructure. Together with magelonids, oweniids have been placed within the Palaeoannelida, a sister group to all remaining annelids. The study of this group may increase our understanding of the early evolution of annelids (including their radiation and diversification). In the current research, the morphology and ulta-anatomy of the head region of Owenia borealis is studied by scanning electron microscopy (SEM), 3D reconstructions, transmission electron microscopy (TEM), and whole-mount immunostaining with confocal laser scanning microscopy. According to SEM, the tentacle apparatus consists of 8–14 branched arms, which are covered by monociliary cells that form a ciliary groove extending along the oral side of the arm base. Each tentacle contains a coelomic cavity with a network of blood capillaries. Monociliary myoepithelial cells of the tentacle coelomic cavity form both the longitudinal and the transverse muscles. The structure of this myoepithelium is intermediate between a simple and pseudo-stratified myoepithelium. Overall, tentacles lack prominent zonality, i.e., co-localization of ciliary zones, neurite bundles, and muscles. This organization, which indicates a non-specialized tentacle crown in O. borealis and other oweniids with tentacles, may be ancestral for annelids. TEM, light, and confocal laser scanning microscopy revealed that the head region contains the anterior nerve center comprising of outer and inner (=circumoral) nerve rings. Both nerve rings are organized as concentrated nerve plexus, which contains perikarya and neurites extending between basal projections of epithelial cells (radial glia). The outer nerve ring gives rise to several thick neurite bundles, which branch and extend along aboral side of each tentacle. Accordingly to their immunoreactivity, both rings of the anterior nerve center could be homologized with the dorsal roots of circumesophageal connectives of the typical annelids. Accordingly to its ultrastructure, the outer nerve ring of O. borealis and so-called brain of other oweniids can not be regarded as a typical brain, i.e. the most anterior ganglion, because it lacks ganglionic structure.

Dio2-CreErt2 Knockin Model to Identify T3-Generating Cells That Express Type 2 Deiodinase in Tissues

Background: Thyroid hormone promotes many developmental and homeostatic functions. Apart from adequate circulating levels, the concentration of the active hormone T3 within tissues may be amplified by type 2 deiodinase (Dio2) by conversion from the precursor T4. Dio2 is critical in auditory development, bone maturation, brain function and control of the hypothalamic-pituitary-thyroid axis. Despite its crucial role, an obstacle to studying Dio2 is that the protein has a short half-life, is at low levels and is often transiently expressed, making it difficult to identify Dio2 in tissues at cellular resolution. Methods: We derived a Dio2-CreERt2 knockin mouse that expresses tamoxifen-dependent Cre recombinase from the endogenous Dio2 gene. When crossed onto Ai6 reporter mice, following tamoxifen treatment, Dio2-CreERt2 expression is detected as fluorescent signal in specific cells in brain regions, pituitary, and other tissues. We showed previously that Dio2 is essential for hearing with rising expression levels in the cochlea prior to onset of hearing. The Dio2-CreERt2 model identified positive cell types in the cochlear spiral ligament, septal divisions and modiolus around the sensory epithelium. Dio2-positive fibrocytes were adjacent to and extended projections around blood capillary networks, the source of T4 supply. Transcriptome analysis of isolated positive cells revealed bone lineage-related origins for many of these cells. Conclusion: The Dio2-CreERt2 model detects Dio2 expression sensitively at cellular resolution. In the cochlea, Dio2-positive cell types reside in vascularized support tissues, suggesting combined endocrine and paracrine-like control of the T3 supply. Analysis of cell origins suggests novel interactions between endocrine and skeletal systems in promoting T3 action required for hearing.

Exploratory study of non-invasive, high-resolution functional macular imaging in subjects with diabetic retinopathy.

To evaluate a high-resolution functional imaging device that yields quantitative data regarding macular blood flow and capillary network features in eyes with diabetic retinopathy (DR). Prospective, cross-sectional comparative case-series in which blood flow velocities (BFVs) and non-invasive capillary perfusion maps (nCPMs) in macular vessels were measured in patients with DR and in healthy controls using the Retinal Functional Imager (RFI) device. A total of 27 eyes of 21 subjects were studied [9 eyes nonproliferative diabetic retinopathy (NPDR), 9 eyes proliferative diabetic retinopathy (PDR) and 9 controls]. All diabetic patients were type 2. All patients with NPDR and 5 eyes with PDR also had diabetic macular edema (DME). The NPDR group included eyes with severe (n=3) and moderate NPDR (n=6), and were symptomatic. A significant decrease in venular BFVs was observed in the macular region of PDR eyes when compared to controls (2.61±0.6 mm/s and 2.92±0.72 mm/s in PDR and controls, respectively, P=0.019) as well as PDR eyes with DME compared to NPDR eyes (2.36±0.51 mm/s and 2.94±1.09 mm/s in PDR with DME and NPDR, respectively, P=0.01). The RFI, a non-invasive imaging tool, provides high-resolution functional imaging of the retinal microvasculature and quantitative measurement of BFVs in visually impaired DR patients. The isolated diminish venular BFVs in PDR eyes compared to healthy eyes and PDR eyes with DME in comparison to NPDR eyes may indicate the possibility of more retinal vein compromise than suspected in advanced DR.

A new model for the emergence of blood capillary networks

We propose a new model for the emergence of blood capillary networks. We assimilate the tissue and extra cellular matrix as a porous medium, using Darcy's law for describing both blood and intersticial fluid flows. Oxygen obeys a convection-diffusionreaction equation describing advection by the blood, diffusion and consumption by the tissue. Discrete agents named capillary elements and modelling groups of endothelial cells are created or deleted according to different rules involving the oxygen concentration gradient, the blood velocity, the sheer stress or the capillary element density. Once created, a capillary element locally enhances the hydraulic conductivity matrix, contributing to a local increase of the blood velocity and oxygen flow. No connectivity between the capillary elements is imposed. The coupling between blood, oxygen flow and capillary elements provides a positive feedback mechanism which triggers the emergence of a network of channels of high hydraulic conductivity which we identify as new blood capillaries. We provide two different, biologically relevant geometrical settings and numerically analyze the influence of each of the capillary creation mechanism in detail. All mechanisms seem to concur towards a harmonious network but the most important ones are those involving oxygen gradient and sheer stress. A detailed discussion of this model with respect to the literature and its potential future developments concludes the paper.

Mechanical adaptation of monocytes in model lung capillary networks

Proper circulation of white blood cells (WBCs) in the pulmonary vascular bed is crucial for an effective immune response. In this branched vascular network, WBCs have to strongly deform to pass through the narrowest capillaries and bifurcations. Although it is known that this process depends on the cell mechanical properties, it is still poorly understood due to the lack of a comprehensive model of cell mechanics and of physiologically relevant experiments. Here, using an in-house microfluidic device mimicking the pulmonary capillary bed, we show that the dynamics of THP1 monocytes evolves along successive capillary-like channels, from a nonstationary slow motion with hops to a fast and smooth efficient one. We used actin cytoskeleton drugs to modify the traffic dynamics. This led us to propose a simple mechanical model that shows that a very finely tuned cortical tension combined with a high cell viscosity governs the fast transit through the network while preserving cell integrity. We finally highlight that the cortical tension controls the steady-state cell velocity via the viscous friction between the cell and the channel walls.

A 3D polydimethylsiloxane microhourglass-shaped channel array made by reflowing photoresist structures for engineering a blood capillary network.

This paper describes an innovative yet straightforward fabrication technique to create three-dimensional microstructures with controllable tapered geometries by combining conventional photolithography and thermal reflow of photoresist. Positive photoresist-based microchannel structures with varying width-to-length ratios were reflowed after their fabrication to generate three-dimensional funnel structures with varying curvatures. A polydimethylsiloxane hourglass-shaped microchannel array was next cast on these photoresist structures, and primary human lung microvascular endothelial cells were cultured in the device to engineer an artificial capillary network. Our work demonstrates that this cost-effective and straightforward fabrication technique has great potential in engineering three-dimensional microstructures for biomedical and biotechnological applications such as blood vessel regeneration strategies, drug screening for vascular diseases, microcolumns for bioseparation, and other fluid dynamic studies at microscale.

Wholly vascularized millimeter-sized engineered tissues by cell-sized microscaffolds.

The in vitro fabrication of wholly vascularized millimeter-sized engineered tissues is still a key challenge in the tissue engineering field. Recently we reported a unique approach ‘sedimentary culture’ using a collagen microfiber (CMF) to fabricate large-scale engineered tissues. The millimeter-sized tissues with high extracellular matrix (ECM) density were easily obtained by centrifugation of cells and CMFs and subsequent cultivation because the CMFs acted as a micrometer-sized scaffold. However, cell distribution in the obtained tissues was not homogeneous because of the different sedimentation velocity of the cells and CMFs because of their size difference.Here we report the fabrication of wholly vascularized millimeter-sized engineered tissues using cell-sized CMFs. To avoid dissolving, vacuum drying was performed at 200 °C for 24 h for thermal crosslinking of primary amine groups of type I collagen. The 200- and 20-μm-sized CMFs (CMF-200 and CMF-20) were obtained by homogenization and subsequent sonication of the crosslinked collagen. Interestingly, the CMF-20 indicated a similar sedimentation velocity with cells because of their same size range, thus uniform millimeter-sized tissue with homogeneous cell distribution was fabricated by the sedimentary culture method. To form a whole blood capillary structure in the tissues, fibronectin (FN) was adsorbed on the surface of CMF-20 to stimulate endothelial cell migration. The distribution of the blood capillary network in 1.6-mm-sized tissues was markedly improved by FN-adsorbed CMF-20 (FN-CMF-20). Sedimentary culture using FN-CMF-20 will create new opportunities in tissue engineering for the in vitro fabrication of wholly vascularized millimeter-sized engineered tissues.

Three-dimensional imaging of spatio-temporal dynamics of small blood capillary network in the cortex based on optical coherence tomography: A review

Optical coherence tomography angiography (OCTA) has emerged as an advanced in vivo imaging modality, which is widely used for the clinic ophthalmology and neuroscience research in the rodent brain cortex among others. Based on the high numerical aperture (NA) probing lens and the motion-corrected algorithms, a high-resolution imaging technique called OCT micro-angiography is applied to resolve the small blood capillary vessels ranging from 5[Formula: see text][Formula: see text]m to 10[Formula: see text][Formula: see text]m in diameter. As OCT-based techniques are recently evolving further from the structural imaging of capillaries toward spatio-temporal dynamic imaging of blood flow in capillaries, here we present a review on the latest techniques for the dynamic flow imaging. Studies on capillary blood flow using these techniques will help us better understand the roles of capillary blood flow for normal functioning of the brain as well as how it malfunctions in diseases.

Deep-learning-based motion-correction algorithm in optical resolution photoacoustic microscopy

In this study, we propose a deep-learning-based method to correct motion artifacts in optical resolution photoacoustic microscopy (OR-PAM). The method is a convolutional neural network that establishes an end-to-end map from input raw data with motion artifacts to output corrected images. First, we performed simulation studies to evaluate the feasibility and effectiveness of the proposed method. Second, we employed this method to process images of rat brain vessels with multiple motion artifacts to evaluate its performance for in vivo applications. The results demonstrate that this method works well for both large blood vessels and capillary networks. In comparison with traditional methods, the proposed method in this study can be easily modified to satisfy different scenarios of motion corrections in OR-PAM by revising the training sets.

Construction of Vascularized Oral Mucosa Equivalents Using a Layer-by-Layer Cell Coating Technology.

There have been many advances in tissue engineering with respect to in vitro and in vivo models of oral mucosa equivalents (OMEs). To apply in vitro reconstructed oral mucosa models to regenerative medicine and alternatives to animal testing, it is necessary to develop the technology of reconstructing different types of oral tissues, such as control of epithelial differentiation and introduction of appendages. We previously reported that functional three-dimensional (3D) tissue models could be quickly constructed by using a layer-by-layer (LbL) cell coating technique that assembles extracellular matrix (ECM) nanofilms to a cell surface. In this study, 3D human OMEs composed of lamina-propria, keratinized or non-keratinized epithelium, and blood capillaries were constructed by using the LbL cell coating technology. Human oral mucosal fibroblasts (HOMFs) were coated with ECM nanofilms and accumulated for the construction of oral mucosal lamina-propria. To construct OMEs with keratinized or non-keratinized epithelium, human oral keratinocytes isolated from gingiva (human oral gingival keratinocytes: HOGKs) or human oral keratinocytes isolated from oral mucosa (human oral mucosal keratinocytes: HOMKs) were used in this study. We further studied the construction of epithelialized OMEs with density- and size-controlled blood capillary networks by using human umbilical vein endothelial cells (HUVECs). It was revealed that these constructions had barrier functions in accordance with their histological characterization. The OMEs with keratinization (K-OMEs) showed higher transepithelial electrical resistance (TEER) values compared with OMEs with non-keratinization (N-OMEs). The constructed epithelialized OMEs with blood capillaries are useful for in vitro/ex vivo research models and regenerative medicine as in oral tissue regeneration. The results suggest that OMEs with oral tissue appendages are more promising alternatives to animal testing and can be applied to the design of in vitro oral models that mimic human tissue organs.

Histological and Ultrastructural Studies on the Intestine of Guntea Loach, Lepidocephalichthys Guntea (Cypriniformes, Cobitidae)

Abstract The cellular organizations of intestine in Lepidocephalichthys guntea (Hamilton, 1822) have been described by light as well as scanning and transmission electron microscopy. The intestine is short and straight like, marked into anterior, middle and posterior region based on mucosal folds, number and size of columnar epithelial cells and mucous cells, thickness of submucosa and muscularis layer. The mucosa of anterior intestine forms high folds, which are lined with compactly arranged columnar epithelial cells and mucous cells. In the middle intestine, folds are pointless whereas the posterior intestine is without folds. The submucosa is formed of thin layer of connective tissue, contained collagen bundles and blood capillaries, comparatively well developed in the posterior intestine. By scanning electron microscopy, outlines of the luminal surface of anterior and middle intestine is embossed with oval or rounded columnar epithelial cells contained densely packed stubby microridges. The posterior intestine has closely set longitudinal folds characterized with minute blood capillaries and columnar epithelial cells having inconspicuous microridges. Ultrastructurally, the mucosal surface of the intestine consists of mucous cells with electron dense granules and columnar epithelial cells having numerous microvilli, mitochondria, endoplasmic reticulum, lysosomes and Golgi body. Cellular components of the anterior and middle intestine participate in the absorption whereas the presence of enormous blood vessels and capillary net work of posterior intestine probably responsible for air breathing.

Solid lipid nanoparticles for targeted delivery of triclosan into skin for infection prevention

Healthcare-associated infections (HAIs) are a concern for health service providers, exacerbated by poor delivery of antimicrobials to target sites within the skin. The dermal route is attractive for local and systemic delivery of drugs, however; permeation, penetration, and access to deeper skin layers are restricted due to the barrier function of the stratum corneum (SC). Solid lipid nanoparticles present several benefits for topical delivery for therapeutic applications, especially via the follicular route. Hair follicles, surrounded by a close network of blood capillaries and dendritic cells, are an important target for delivery of antimicrobials and present a unique microbial nidus for endogenous infections in situations where the barrier is disrupted, such as after surgery, for example, triclosan, a broad-spectrum antimicrobial agent, was encapsulated into nanoparticles using glyceryl behenate and glyceryl palmitostearate (GP) solid lipids, and incorporating Transcutol P, a known permeation enhancer at different ratios. Optimised formulation was stable over 90 d and in vitro permeation studies using full thickness porcine ear skin showed that the lipid-based nanoparticles enhanced delivery of triclosan into the skin and could direct the agent towards hair follicles, indicating their potential as a carrier system for antiseptic dermal delivery.

Image enhancement effect on inter and intra-observer reliability of nailfold capillary assessment

BackgroundNailfold capillaroscopy (NC) is a diagnostic imaging technique that is used to assess the blood capillary network in the nailfold area. NC is routinely used for patients with microcirculation problems, such as systemic sclerosis and other connective tissue diseases. Experts commonly use subjective evaluation as a reference point in images of nailfold video capillaroscopy, so it is important to reduce the inherent ambiguities in human judgment and diagnosis. Image quality is an important factor that affects measurement error and assessment time of NC images. ObjectiveIn this study, a new image enhancement technique was introduced and evaluated subjectively. MethodsIn total, 475 nailfold video capillaroscopy images from 18 healthy subjects and 41 systemic lupus erythematosus patients were used. The images were randomly divided into two sets, one each with 275 and 200. Eight independent observers who were familiar with the capillaroscopy technique participated in this study. The set of 275 images was evaluated by three observers with the forced–choice pairwise comparison method. Elliptic broken line (EBL) was used to count the number of capillaries. The intra- and inter-observer reliability of the original and enhanced images was evaluated on 200 images by five observers. ResultExcept for eight images, all observers preferred the enhanced images in the visual quality comparison method. The intra-class correlation coefficient (ICC) of intra- and inter-observer reliability increased from 0.76–0.84 to 0.82–0.89, respectively, when using the enhancement method. ConclusionBy improving the image quality, more capillary details will be visible, and an observer can document more details that may not be visible in the original image and can do so more efficiently.

Intermittent Pili-Mediated Forces Fluidize Neisseria meningitidis Aggregates Promoting Vascular Colonization

Neisseria meningitidis, a bacterium responsible for meningitis and septicemia, proliferates and eventually fills the lumen of blood capillaries with multicellular aggregates. The impact of this aggregation process and its specific properties are unknown. We first show that aggregative properties are necessary for efficient infection and study their underlying physical mechanisms. Micropipette aspiration and single-cell tracking unravel unique features of an atypical fluidized phase, with single-cell diffusion exceeding that of isolated cells. A quantitative description of the bacterial pair interactions combined with active matter physics-based modeling show that this behavior relies on type IV pili active dynamics that mediate alternating phases of bacteria fast mutual approach, contact, and release. These peculiar fluid properties proved necessary to adjust to the geometry of capillaries upon bacterial proliferation. Intermittent attractive forces thus generate a fluidized phase that allows for efficient colonization of the blood capillary network during infection.