Microvascular Structures Research Articles

Lauric acid provides neuroprotection against oxidative stress in mouse model of hyperglycaemic stroke

During ischemic stroke, higher glucose level linked worse outcomes were reported even in patients without pre-existing diabetes. Evidence suggest that such worse stroke outcomes were mainly due to production of reactive, toxic glucose metabolites that expands oxidative damage inside the brain. As a consequence of high oxidative stress, microvasculature structures and tight junctions compromised their functionally, infarct volume expands and brain edema exacerbates. In a mouse model of ischemic stroke with induced acute hyperglycaemia, Lauric acid (LA) as a natural saturated fatty acid demonstrated neuroprotection by attenuating infarct volume and brain edema. In addition, in the ipsilateral hyperglycaemic brain, the LA significantly increased the expression of tight junction representative protein (occludin) as well as anti-oxidative markers; Manganese superoxide dismutase (Mn) SOD, Extracellular superoxide dismutase (Ec-SOD) and nuclear factor-erythroid factor 2-related factor 2 (Nrf2) in the ipsilateral region against hyperglycemic ischemic stroke. LA treated animals showed a significant reduction in the production of lipid peroxidation products (4-HNE) in the microvascular structures, maintained the blood brain barrier (BBB) integrity. LA linked neuroprotective outcomes were further confirmed by behavioral tests, where functional outcomes and motor coordination were improved significantly. Furthermore, LA treatment enhanced food intake, decreased mortality rate, and net body weight loss. Conclusively, LA modulated ischemic insult exacerbated by hyperglycemia and provided neuroprotection.

Dual-path multi-scale context dense aggregation network for retinal vessel segmentation

There has been steady progress in the field of deep learning-based blood vessel segmentation. However, several challenging issues still continue to limit its progress, including inadequate sample sizes, the neglect of contextual information, and the loss of microvascular details. To address these limitations, we propose a dual-path deep learning framework for blood vessel segmentation. In our framework, the fundus images are divided into concentric patches with different scales to alleviate the overfitting problem. Then, a Multi-scale Context Dense Aggregation Network (MCDAU-Net) is proposed to accurately extract the blood vessel boundaries from these patches. In MCDAU-Net, a Cascaded Dilated Spatial Pyramid Pooling (CDSPP) module is designed and incorporated into intermediate layers of the model, enhancing the receptive field and producing feature maps enriched with contextual information. To improve segmentation performance for low-contrast vessels, we propose an InceptionConv (IConv) module, which can explore deeper semantic features and suppress the propagation of non-vessel information. Furthermore, we design a Multi-scale Adaptive Feature Aggregation (MAFA) module to fuse the multi-scale feature by assigning adaptive weight coefficients to different feature maps through skip connections. Finally, to explore the complementary contextual information and enhance the continuity of microvascular structures, a fusion module is designed to combine the segmentation results obtained from patches of different sizes, achieving fine microvascular segmentation performance. In order to assess the effectiveness of our approach, we conducted evaluations on three widely-used public datasets: DRIVE, CHASE-DB1, and STARE. Our findings reveal a remarkable advancement over the current state-of-the-art (SOTA) techniques, with the mean values of Se and F1 scores being an increase of 7.9% and 4.7%, respectively. The code is available at https://github.com/bai101315/MCDAU-Net.

Ultrasound Localization Microscopy Using Deep Neural Network

Noninvasive imaging of microvascular structures in deep tissues provides morphological and functional information for clinical diagnosis and monitoring. Ultrasound Localization Microscopy (ULM) is an emerging imaging technique that can generate microvascular structures with subwavelength diffraction resolution. However, the clinical utility of ULM is hindered by technical limitations, such as long data acquisition time, high microbubble concentration, and inaccurate localization. In this paper, we propose a Swin transformer-based neural network to perform end-to-end mapping to implement microbubble localization. The performance of the proposed method was validated using synthetic and in vivo data using different quantitative metrics. The results indicate that our proposed network can achieve higher precision and better imaging capability than previously used methods. Furthermore, the computational cost of processing per frame is three to four times faster than traditional methods, which makes the real-time application of this technique feasible in the future.

Deep optoacoustic localization microangiography of ischemic stroke in mice

Super-resolution optoacoustic imaging of microvascular structures deep in mammalian tissues has so far been impeded by strong absorption from densely-packed red blood cells. Here we devised 5 µm biocompatible dichloromethane-based microdroplets exhibiting several orders of magnitude higher optical absorption than red blood cells at near-infrared wavelengths, thus enabling single-particle detection in vivo. We demonstrate non-invasive three-dimensional microangiography of the mouse brain beyond the acoustic diffraction limit (<20 µm resolution). Blood flow velocity quantification in microvascular networks and light fluence mapping was also accomplished. In mice affected by acute ischemic stroke, the multi-parametric multi-scale observations enabled by super-resolution and spectroscopic optoacoustic imaging revealed significant differences in microvascular density, flow and oxygen saturation in ipsi- and contra-lateral brain hemispheres. Given the sensitivity of optoacoustics to functional, metabolic and molecular events in living tissues, the new approach paves the way for non-invasive microscopic observations with unrivaled resolution, contrast and speed.

Diagnosing Systemic Disorders with AI Algorithms Based on Ocular Images.

The advent of artificial intelligence (AI), especially the state-of-the-art deep learning frameworks, has begun a silent revolution in all medical subfields, including ophthalmology. Due to their specific microvascular and neural structures, the eyes are anatomically associated with the rest of the body. Hence, ocular image-based AI technology may be a useful alternative or additional screening strategy for systemic diseases, especially where resources are scarce. This review summarizes the current applications of AI related to the prediction of systemic diseases from multimodal ocular images, including cardiovascular diseases, dementia, chronic kidney diseases, and anemia. Finally, we also discuss the current predicaments and future directions of these applications.

Microvascular Morphology MR Imaging Using Relaxation Rate Change with Gadolinium‐Based Contrast Agent in Patients with Alzheimer’s Disease

AbstractBackgroundConventional MRI techniques cannot demonstrate microvascular alterations in mild and moderate Alzheimer’s disease (AD). Thus, the diagnosis of microvascular pathology commonly relies on postmortem. The purpose of this study was to evaluate alterations of microvascular structures in patients with AD using a 3T clinical MRI system with a commercially available contrast agent.MethodEleven patients with AD and 11 non‐AD controls were included in this cross‐sectional prospective study. R2 and R2* relaxation rate changes (∆R2 and ∆R2*) before and after a Gd‐based contrast agent injection were calculated from images obtained with a multi‐echo turbo spin‐echo sequence and multi‐echo gradient‐echo sequence to obtain microvascular index maps of blood volume fraction (BVf), mean vessel diameter (mVD), vessel size index (VSI), mean vessel density (Q), and microvessel‐weighted imaging (MvWI). Two‐sample t‐test was used to compare those values between the two groups. Correlation analysis was performed to evaluate the relationship between those values and the mini‐mental state examination (MMSE) score.ResultCompared with the non‐AD group, BVf was significantly increased in the AD group at the parahippocampal gyrus (p = 0.033), gray matter area (p = 0.008), and white matter area (p = 0.010). MvWI was also significantly increased in the AD group at the hippocampus (p = 0.010), parahippocampal gyrus (p = 0.010), precuneus (p = 0.045), and globus pallidus (p = 0.008). Both mVD and VSI were only significantly decreased in the AD group at the white matter hyperintensity (WMHI) area (p = 0.045 with mVD; p = 0.033 with VSI). BVf and MvWI were significantly positively correlated with age at some areas. VSI was significantly positively correlated with MMSE at caudate (rho = 0.475/ p = 0.030) and corpus callosum (rho = 0.435/ p = 0.049).ConclusionMicrovascular index was a useful non‐invasive method to evaluate microvascular morphology alteration. The microvascular morphology of AD was manifested as vessel constriction and increasing blood volume fraction. It was associated with MMSE score.

Effect of oral contraceptive pills on the retinal microvascular structure: An optical coherence tomography angiography study

BackgroundTo investigate the effect of oral contraceptive pills (OCPs) on foveal avascular zone (FAZ), peripapillary capillary plexus, and superficial and deep capillary plexus (SCP and DCP) measurements. MethodsThis cross-sectionally designed study included 32 healthy female participants using OCPs (3 mg drospirenone and 0.03 mg ethinylestradiol) for at least one year for contraception and 32 healthy controls that did not use any drugs. All subjects were evaluated using optical coherence tomography angiography (OCTA). Using OCTA, the measurements of SCP, DCP, radial peripapillary capillary (RPC) vessel density; FAZ area and perimeter; acircularity index (AI); and foveal density (FD) were undertaken. Each participant's measurements were taken while they were in the follicular phase of their menstrual cycles (day 3). ResultsAge and body mass index did not significantly differ between the groups (p = 0.56 and p = 0.15, respectively). The DCP vessel densities in all the regions were lower in the OCP group (p<0.05 for all). The vessel densities of SCP and RPC, FAZ area and perimeter, AI, and FD were similar between the two groups (p>0.05 for all). ConclusionWe determined that the DCP vessel density was reduced in women using this drug. OCPs can cause changes in retinal microvascular structures. Therefore, OCTA can be used in the follow-up of healthy women using OCP.

Assessment of Transarterial Chemoembolization Using Super-resolution Ultrasound Imaging and a Rat Model of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is a highly prevalent form of liver cancer diagnosed annually in 600,000 people worldwide. A common treatment is transarterial chemoembolization (TACE), which interrupts the blood supply of oxygen and nutrients to the tumor mass. The need for repeat TACE treatments may be assessed in the weeks after therapy with contrast-enhanced ultrasound (CEUS) imaging. Although the spatial resolution of traditional CEUS has been restricted by the diffraction limit of ultrasound (US), this physical barrier has been overcome by a recent innovation known as super-resolution US (SRUS) imaging. In short, SRUS enhances the visible details of smaller microvascular structures on the 10 to 100 µm scale, which unlocks a host of new clinical opportunities for US. In this study, a rat model of orthotopic HCC is introduced and TACE treatment response (to a doxorubicin-lipiodol emulsion) is assessed using longitudinal SRUS and magnetic resonance imaging (MRI) performed at 0, 7 and 14 d. Animals were euthanized at 14 d for histological analysis of excised tumor tissue and determination of TACE response, that is, control, partial response or complete response. CEUS imaging was performed using a pre-clinical US system (Vevo 3100, FUJIFILM VisualSonics Inc.) equipped with an MX201 linear array transducer. After administration of a microbubble contrast agent (Definity, Lantheus Medical Imaging), a series of CEUS images were collected at each tissue cross-section as the transducer was mechanically stepped at 100 μm increments. SRUS images were formed at each spatial position, and a microvascular density metric was calculated. Microscale computed tomography (microCT, OI/CT, MILabs) was used to confirm TACE procedure success, and tumor size was monitored using a small animal MRI system (BioSpec 3T, Bruker Corp.). Although there were no differences at baseline (p > 0.15), both microvascular density levels and tumor size measures from the complete responder cases at 14 d were considerably lower and smaller, respectively, than those in the partial responder or control group animals. Histological analysis revealed tumor-to-necrosis levels of 8.4%, 51.1% and 100%, for the control, partial responder and complete responder groups, respectively (p < 0.005). SRUS imaging is a promising modality for assessing early changes in microvascular networks in response to tissue perfusion-altering interventions such as TACE treatment of HCC.

OCTA evaluation of posterior ocular blood flow in patients after COVID-19 infection without pneumonia

We aimed to demonstrate the changes in optic nerve and retinal microvascular structures with Optical Coherence Tomography Angiography (OCTA) in patients who had COVID-19 infection. Prospective study. Retina, choroid and optic nerve head microvascular flow and vascular densities of both groups were measured by OCTA. OCTA measurements of 122 right eyes of a total of 122 patients, including 72 patients in the COVID-19 group and 50 patients in the control group, were included in the study. The Deep Capillary Plexus (DCP) Flow Area (FA) in the COVID-19 group was 1.42±0.23mm2, in the control group was 1.50±0.15mm2, Choriocapillary Plexus FA was 1.89±0.04mm2 in the COVID-19 group, was 1.91±0.05mm2 in the control group, and a statistically significant difference was found between the two groups (respectively; P=0.03; P=0.02). DCP Whole Vascular density (VD) values were 56.76±4.16% in the COVID-19 group, 58.28±3.88% in the control group; difference between the values was statistically significant (P=0.04). There was no statistically significant difference between the two groups in terms of optic nerve head flow areas and other parameters examined according to quadrants. Results show that the retinal microcirculation in subjects with mild disease is affected. Even if the disease is mild, patients may need to be followed up for retinal changes that may develop in the future.

Speed-resolved perfusion imaging using multi-exposure laser speckle contrast imaging and machine learning.

Laser speckle contrast imaging (LSCI) gives a relative measure of microcirculatory perfusion. However, due to the limited information in single-exposure LSCI, models are inaccurate for skin tissue due to complex effects from e.g. static and dynamic scatterers, multiple Doppler shifts, and the speed-distribution of blood. It has been demonstrated how to account for these effects in laser Doppler flowmetry (LDF) using inverse Monte Carlo (MC) algorithms. This allows for a speed-resolved perfusion measure in absolute units %RBC × mm/s, improving the physiological interpretation of the data. Until now, this has been limited to a single-point LDF technique but recent advances in multi-exposure LSCI (MELSCI) enable the analysis in an imaging modality. To present a method for speed-resolved perfusion imaging in absolute units %RBC × mm/s, computed from multi-exposure speckle contrast images. An artificial neural network (ANN) was trained on a large simulated dataset of multi-exposure contrast values and corresponding speed-resolved perfusion. The dataset was generated using MC simulations of photon transport in randomized skin models covering a wide range of physiologically relevant geometrical and optical tissue properties. The ANN was evaluated on in vivo data sets captured during an occlusion provocation. Speed-resolved perfusion was estimated in the three speed intervals 0 to , 1 to , and , with relative errors 9.8%, 12%, and 19%, respectively. The perfusion had a linear response to changes in both blood tissue fraction and blood flow speed and was less affected by tissue properties compared with single-exposure LSCI. The image quality was subjectively higher compared with LSCI, revealing previously unseen macro- and microvascular structures. The ANN, trained on modeled data, calculates speed-resolved perfusion in absolute units from multi-exposure speckle contrast. This method facilitates the physiological interpretation of measurements using MELSCI and may increase the clinical impact of the technique.

Free fatty acids induce coronary microvascular dysfunction via inhibition of the AMPK/KLF2/eNOS signaling pathway.

Increased levels of serum free fatty acids (FFAs) are closely associated with microvascular dysfunction. In our previous study, a coronary microvascular dysfunction (CMD) model was successfully established via lipid infusion to increase the levels of serum FFAs in mice. However, the underlying mechanisms remained poorly understood. Therefore, the aim of the present study was to explore the mechanism underlying FFA‑induced CMD. A CMD mouse model was established via lipid combined with heparin infusion for 6h to increase the concentration of serum FFAs. Following the establishment of the model, the coronary flow reserve (CFR), extent of leukocyte activation and cardiac microvascular structures were assessed in the mice. Cardiac microvascular endothelial cells (CMECs) were treated with different concentrations of palmitic acid and cell viability was evaluated. Changes in the expression levels of AMP‑activated protein kinase (AMPK), Krüppel‑like factor 2 (KLF2) and endothelial nitric oxide synthase (eNOS) were identified by immunohistochemical and western blot analyses. Experiments using AMPK activator, KLF2 overexpression plasmid, small interfering RNAs and nicorandil were subsequently designed to investigate the potential involvement of the AMPK/KLF2/eNOS signaling pathway. These experiments revealed that FFAs could induce CMD in mice, which was characterized by reduced CFR (1.89±0.37 vs. 2.74±0.30) and increased leukocyte adhesion (4,350±1,057.5 vs. 11.8±5.4cells/mm2) compared with the control mice. CD11b expression and intracellular reactive oxygen species (ROS) levels were increased in CMD model mice compared with control mice. Serum TNF‑α and IL‑6 levels were higher in the model group than in the control group. Transmission electron microscopy revealed that CMECs in heart tissues of model mice were severely swollen. In addition, palmitic acid decreased CMEC viability and increased ROS production in a dose‑dependent manner. Notably, the AMPK/KLF2/eNOS signaling pathway was demonstrated to be suppressed by FFAs both invivo and invitro. Activation of this axis with AMPK activator, KLF2 overexpression plasmid or nicorandil restored the CFR in CMD model mice, inhibited oxidative stress and increased CMEC viability. Taken together, the results of the present study demonstrated that FFAs could induce CMD via inhibition of the AMPK/KLF2/eNOS signaling pathway, whereas activation of this pathway led to the alleviation of FFA‑induced CMD, which may be a therapeutic option for CMD.

Abstract P2-15-06: Light-based bioprinting of vascularized fatty tissue for the development of biological breast implants

Abstract Although multi-modal and ablative treatment strategies for breast cancer are becoming more popular, partial or full mastectomy is still a most common intervention. While proving to be effective in curing the cancer, it leaves the patient disfigured, which can pose a considerable psychological burden. Hence, reconstructive breast surgery is continuing to increase in importance as it offers the possibility to at least partially restore the original look of the breast. Autologous adipose tissue transfer techniques, amongst them the surgical gold standard flap surgery and liposuction-based fat grafting, present natural solutions with the patient’s own tissue, but either produce a considerable secondary wound or are insufficient to address larger defects. Synthetic solutions like silicone implants on the other hand can also be used to fill large defects and only require minimal surgical intervention, but suffer from foreign body-associated complications like fibrosis. Tissue engineering biological breast implants of arbitrary size with no or minor insult to the patient’s body and excellent biocompatibility, exclusively using biomaterials and patient-specific adipose cells, could open a new chapter for breast reconstruction. Such an approach has become an intriguing possibility through the rise of powerful biofabrication technologies in the last two decades, namely three-dimensional bioprinting. Bioprinting marries additive manufacturing with biomaterials and living cells with the aim to create life-mimicking tissues and organs, and while it is mainly focused on research today, current endeavors push for application in regenerative medicine. In our work we sought to fabricate adipose tissue constructs using light-based bioprinting to investigate the potential application of such technology for future reconstructive breast surgery. The engineered tissue blocks were designed to mimic native human adipose tissue by comprising fat, connective tissue and endothelial cells. Being specifically placed by the bioprinter within extracellular matrix-like hydrogels, the cells were cultured in vitro and differentiated within the cultivation time of up to 4 weeks to form an adipose-like tissue. Live imaging showed high viability of bioprinted tissue blocks over the whole cultivation period and pronounced cellular reorganization. Confocal and histological analyses revealed a dynamic tissue maturation process with increasing proportions of differentiated adipocytes, ECM remodeling and formation of microvascular structures. In conclusion, the obtained results gave proof to the concept of engineering adipose tissue through bioprinting and could lay the groundwork for the development of all-natural biological breast implants. Citation Format: Nina Hedemann, Alexander Thomas, Nils Tribian, Lutz Kloke, Dirk Bauerschlag, Marion van Mackelenbergh. Light-based bioprinting of vascularized fatty tissue for the development of biological breast implants [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-15-06.

Retinal Vascular Tortuosity Index Change after Idiopathic Epiretinal Membrane Surgery: Does Internal Limiting Membrane Peeling Affect Retinal Vascular Tortuosity?

Idiopathic epiretinal membrane (iERM) surgery is one of the most commonly performed vitreoretinal surgeries, and the issue of internal limiting membrane (ILM) peeling in ERM surgery is still controversial. The aims of this study are to evaluate the changes in retinal vascular tortuosity index (RVTI) after pars plana vitrectomy for the iERM using optical coherence tomography angiography (OCTA) and to assess whether ILM peeling has an additional effect on RVTI reduction. This study included25 eyes of 25 iERM patients who underwent ERM surgery. The ERM was removed without ILM peeling in 10 eyes (40.0%), and the ILM was peeled in addition to the ERM in 15 eyes (60.0%). The existence of the ILM after ERM peeling was checked with second staining in all eyes. Best corrected visual acuity (BCVA) and 6 × 6 mm en-face OCTA images were recorded before surgery and at the first month postoperatively. A skeleton model of the retinal vascular structure was created following Otsu binarization of en-face OCTA images using ImageJ software (1.52U). RVTI was calculated as the ratio of each vessel length to its Euclidean distance on the skeleton model using the Analyze Skeleton plug-in. The mean RVTI declined from 1.220 ± 0.017 to 1.201 ± 0.020 (p = 0.036) in eyes with ILM peeling and from 1.230 ± 0.038 to 1.195 ± 0.024 in eyes without ILM peeling (p = 0.037). There was no difference between the groups in terms of postoperative RVTI (p = 0.494). A statistically significant correlation was found between postoperative RVTI and postoperative BCVA (rho = 0.408, p = 0.043). The RVTI is an indirect indicator of the traction created by the iERM on retinal microvascular structures, and it was effectively reduced after iERM surgery. The postoperative RVTIs were similar in cases who underwent iERM surgery with or without ILM peeling. Therefore, ILM peeling may not have an additive effect on the loosening of microvascular traction and thus may be reserved for recurrent ERM surgeries.

In Vivo Imaging of Lipid Droplets and Oxygen Status in Hepatic Tissues of Nonalcoholic Fatty Liver Model Mice Using a Lipophilic Ir(III) Complex

Nonalcoholic fatty liver disease (NAFLD) is becoming common worldwide. In pathophysiological studies of NAFLD, an in vivo optical probe that enables visualization of lipid droplets (LDs) and imaging of oxygen status in hepatic tissues simultaneously would be very useful. Here, we present the phosphorescent Ir(III) complex BTP ((btp)2Ir(acac) (btp = benzothienylpyridine, acac = acetylacetone)) as the first probe that meets this requirement. BTP was efficiently taken up into cultured 3T3-L1 adipocytes and selectively accumulated into LDs. Quantifying oxygen levels in LDs based on the phosphorescence lifetime of BTP allowed us to track changes in cellular oxygen tension after treatment with metabolic stimulants. Phosphorescence lifetime imaging microscopy combined with intravenously administered BTP in mice enabled specific visualization of LDs in hepatic lobules and simultaneous imaging of the oxygen gradient that decreased from the portal vein (PV) to the central vein (CV). NAFL model mice were created by feeding a high-fat diet (HFD) to mice for 3 or 7 days. The mice fed an HFD showed a marked increase in the amount and size of LDs in hepatocytes compared with those fed a normal diet, leading to abnormal microvascular structures. In addition, HFD-fed mice also exhibited reduced oxygen tension in areas other than the CV. Multicolor imaging with the LD-accumulated oxygen probe BTP and vasculature-staining FITC-lectin suggested that structural distortions of the sinusoidal microvasculature caused by enlarged LDs were associated with partial hypoxia in NAFL.

Retinal microvascular markers in type 2 diabetes subphenotypes and latent autoimmune diabetes of adults.

To estimate if newly diagnosed patients with different subphenotypes of type 2 diabetes (T2DM) or latent autoimmune diabetes of adults (LADA) differ with respect to subclinical retinal microvascular structure or diabetic retinopathy (DR). This population-based, cross-sectional study of 340 patients (675 eyes) classified patients with recently diagnosed T2DM in different subphenotypes according to beta cell function and insulin sensitivity in to; classical (n=218), hyperinsulinaemic (n=86), insulinopenic (n=20), or LADA (n=16). Retinal 6-field images were graded according to the International Clinical DR Severity Scale by a retinal expert. Retinal microvascular structures were analysed in eyes by a semiautomatic software. Median age and duration of diabetes were 58.1 (49.9; 65.5) and 0.9 (0.5; 2.4) years, respectively, and 56.8% were male. In a multivariate linear mixed model regression analysis of eyes without DR (n=570), there was no statistically significant difference in retinal venular or arteriolar width between subtypes and patients with classical T2DM. In addition, eyes from different subphenotypes did not differ according to vessel density, tortuosity or fractal dimension. In a multivariate logistic regression model adjusted for age, sex, HbA1c, diabetes duration, body mass index, mean arterial blood pressure and history of cardiovascular disease, there was a tendency towards persons with hyperinsulinaemic T2DM to be more likely to have DR (OR 1.97, 95% CI 0.95; 4.09) compared to classical T2DM. We found no difference in retinal microvascular structure in patients with newly diagnosed subtypes of T2DM. However, DR may be more prevalent in newly diagnosed patients with hyperinsulinaemic T2DM compared to individuals with classical T2DM.

The impact of obesity on ocular hemodynamics and choroidal thickness.

Obesity affects microvascular structures. The effect of obesity on the ocular vascular system can be evaluated by changes in the choroidal thickness (CT) and retrobulbar blood flow (RBF). To evaluate the CT and RBF parameters in obese patients with various body mass index (BMI) values and compare these parameters with normal weight, healthy subjects. A prospective study. The study included 102 eyes of 102 female patients. Patients were divided into three groups according to BMI as group 1 with a BMI of 18.5-24.99 (n = 32), normal weight group; group 2 with a BMI of 30-34.99 (n = 35), as obese class I; and group 3 with a BMI of 35-39.99 (n = 35), as obese class II. The peak systolic velocity (PSV), end-diastolic velocity (EDV), resistive index, and pulsatility index values of the central retinal artery (CRA) and ophthalmic artery (OA) were evaluated with color Doppler ultrasonography. CT was measured at the subfoveal area and at 500-µm intervals nasal and temporal to the fovea up to a distance of 1500 µm by using the enhanced depth imaging technique of optical coherence tomography. Intraocular pressure (IOP) was measured with a Goldmann applanation tonometry. There was a significant difference in IOP values within the groups with the highest values in group 3 (17.6 ± 2.1 mmHg) and the lowest in group 1 (12.4 ± 1.7 mmHg). The CT in groups 2 and 3 was found to be statistically significantly lower than that in group 1 at all measurement points (p < 0.001). There was a statistically significant negative correlation between CT at all measurement points and BMI (p < 0.001). The mean CRA PSV, EDV, and OA EDV values were statistically significantly lower in each obese group than those values in group 1 (p < 0.001). The OA PSV values were significantly lower in group 3 (36.5 ± 5.9 cm/s) than those in group 2 (43.8 ± 4 cm/s) and group 1 (44.6 ± 5.2 cm/s) (p < 0.001). Also, significant associations were found between BMI and CRA PSV, CRA EDV, and OA PSV values (p < 0.001). Obesity may predispose to eye pathologies by changing the ocular vascular circulation.

Microvascular morphology alteration using relaxation rate change with gadolinium-based magnetic resonance imaging contrast agent in patients with Alzheimer’s disease

Conventional magnetic resonance imaging (MRI) techniques cannot demonstrate microvascular alterations in mild Alzheimer's disease (AD). Thus, the diagnosis of microvascular pathology commonly relies on postmortem. The purpose of this study was to evaluate alterations of microvascular structures in patients with AD using a 3T clinical MRI system with a commercially available contrast agent. Eleven patients with AD and 11 cognitively normal (CN) controls were included in this cross-sectional prospective study. R2 and R2* relaxation rate changes (∆R2 and ∆R2*) before and after a Gadolinium (Gd)-based contrast agent injection were calculated from images obtained with a multi-echo turbo spin-echo sequence and multi-echo gradient-echo sequence to obtain microvascular index maps of blood volume fraction (BVf), mean vessel diameter (mVD), vessel size index (VSI), mean vessel density (Q), and microvessel-weighted imaging (MvWI). Two-sample t-test was used to compare those values between the two groups. Correlation analysis was performed to evaluate the relationship between those values and age. BVfs at the corpus callosum and at the thalamus were significantly increased in the AD group (P=0.024 and P=0.005, respectively). BVf at the gray matter (P=0.020) and white matter area (P=0.012) were also significantly increased in the AD group compared with the CN group. MvWIs at the hippocampus and parahippocampal gyrus were significantly increased in the AD group compared with the CN group (P=0.020 and P=0.006, respectively). Voxel-based analysis showed both mVD and VSI were significantly decreased at the prefrontal lobe in the AD group. Q were not significant difference between CN and AD groups. MvWI were significantly positively correlated with age. Microvascular index was a useful non-invasive method to evaluate microvascular morphology alteration. The microvascular morphology of AD was manifested as increasing BVf and microvessel-weighted.

Insight into microvascular adaptive alterations in the Glisson system of biliary atresia after Kasai portoenterostomy using X-ray phase-contrast CT.

To investigate microvascular alterations in the Glisson system of biliary atresia (BA) patients after Kasai portoenterostomy (KP) using three-dimensional (3D) virtual histopathology based on X-ray phase-contrast CT (PCCT). Liver explants from BA patients were imaged using PCCT, and 32 subjects were included and divided into two groups: KP (n = 16) and non-KP (n = 16). Combined with histological analysis and 3D visualization technology, 3D virtual histopathological assessment of the biliary, arterial, and portal venous systems was performed. According to loop volume ratio, 3D spatial density, relative surface area, tortuosity, and other parameters, pathological changes of microvasculature in the Glisson system were investigated. In the non-KP group, bile ducts mostly manifested as radial multifurcated hyperplasia and twisted into loops. In the KP group, the bile duct hyperplasia was less, and the loop volume ratio of bile ducts decreased by 13.89%. Simultaneously, the arterial and portal venous systems presented adaptive alterations in response to degrees of bile duct hyperplasia. Compared with the non-KP group, the 3D spatial density of arteries in the KP group decreased by 3.53%, and the relative surface area decreased from 0.088 ± 0.035 to 0.039 ± 0.015 (p < .01). Deformed portal branches gradually recovered after KP, with a 2.93% increase in 3D spatial density and a decrease in tortuosity from 1.17 ± 0.06 to 1.14 ± 0.04 (p < .01) compared to the non-KP group. 3D virtual histopathology via PCCT clearly reveals the microvascular structures in the Glisson system of BA patients and provides key insights into the morphological mechanism of microvascular adaptation induced by biliary tract dredging after KP in BA disease. • 3D virtual histopathology via X-ray phase-contrast computed tomography clearly presented the morphological structures and pathological changes of microvasculature in the Glisson system of biliary atresia patients. • The morphological alterations of microvasculature in the Glisson system followed the competitive occupancy mechanism in the process of biliary atresia.

Identifying a sublingual triangle as the ideal site for assessment of sublingual microcirculation

The sublingual mucosa is a commonly used intraoral location for identifying microcirculatory alterations using handheld vital microscopes (HVMs). The anatomic description of the sublingual cave and its related training have not been adequately introduced. The aim of this study was to introduce anatomy guided sublingual microcirculatory assessment. Measurements were acquired from the floor of the mouth using incident dark-field (IDF) imaging before (T0) and after (T1) sublingual cave anatomy instructed training. Instructions consists of examining a specific region of interested identified through observable anatomical structures adjacent and bilaterally to the lingual frenulum which is next to the sublingual papilla. The anatomical location called the sublingual triangle, was identified as stationed between the lingual frenulum, the sublingual fold and ventrally to the tongue. Small, large, and total vessel density datasets (SVD, LVD and TVD respectively) obtained by non-instructed and instructed measurements (NIN (T0) and IM (T1) respectively) were compared. Microvascular structures were analyzed, and the presence of salivary duct-related microcirculation was identified. A total of 72 video clips were used for analysis in which TVD, but not LVD and SVD, was higher in IM compared to NIM (NIM vs. IM, 25 ± 2 vs. 27 ± 3 mm/mm2 (p = 0.044), LVD NIM vs. IM: 7 ± 1 vs. 8 ± 1mm/mm2 (p = 0.092), SVD NIM vs. IM: 18 ± 2 vs. 20 ± 3 mm/mm2 (p = 0.103)). IM resulted in microcirculatory assessments which included morphological properties such as capillaries, venules and arterioles, without salivary duct-associated microcirculation. The sublingual triangle identified in this study showed consistent network-based microcirculation, without interference from microcirculation associated with specialized anatomic structures. These findings suggest that the sublingual triangle, an anatomy guided location, yielded sublingual based measurements that conforms with international guidelines. IM showed higher TVD values, and future studies are needed with larger sample sizes to prove differences in microcirculatory parameters.

Engineering Vascular Self‐Assembly by Controlled 3D‐Printed Cell Placement

AbstractNutrient supply via a functional vasculature is essential during regenerative processes, tissue growth, and homeostasis. 3D bioprinting offers the opportunity to engineer vascularized constructs by combining cells and biocompatible materials in specifically designed fashions. However, the complexity of microvascular dynamic networks can hardly be recapitulated yet, even by sophisticated 3D manufacturing. Ideally, the natural organizational competences of endothelial cells will be harnessed such that engineered vascular networks self‐assemble to form complex, controllable microvascular patterns. Here, a bioengineering approach is presented to control microvascular structure formation and to steer cellular self‐assembly of endothelial and supporting cells within a multi‐material stereolithographic 3D bioprinting concept. Bioengineered vascularized constructs are generated by controlled cell deposition in an enzymatically degradable or a non‐degradable material. In vitro, the microvascular structures are regulated in distribution, network orientation, vessel length and branching behavior and developed lumen, signs of vascular stabilization and an interconnected vascular network including anastomosis. This novel biofabrication approach demonstrates the capability to control microvascular network formation by using cellular and spatial cues allowing the generation of distinctly yet precisely vascularized constructs. Such novel approach of controlled microvascular formation may play a fundamental role in the development of vascularized implants or in vitro screening models.