Microvasculature Density Research Articles

Retinal morphology across the menstrual cycle: insights from the UK Biobank

Oestradiol and progesterone levels are higher in menstruating women than men of the same age, and their receptors are present in their neurosensory retina and retinal pigment epithelium. However, the impact of this hormonal environment on retinal physiology in women remains unclear. Using self-reported menstrual cycle phases as a surrogate for fluctuating hormonal levels, we investigated associations with retinovascular indices on colour fundus photograph and retinal thickness in optical coherence tomography across regularly menstruating women in the UK Biobank. We found no differences in retinal thickness across the cycle; however, vessel density, arteriolar and venular, and fractal dimension were higher in the luteal phase than follicular. The calibre of the central retinal vessels did not differ. This study suggests that the menstrual cycle phase might be associated with retinal microvasculature density in non-invasive imaging. It raises awareness for this understudied area, providing insights into neuroscience fields and epidemiological studies.

Retinal morphology and microvasculature density alterations in generalized anxiety disorders

Purpose This study evaluates morphological alterations in the macula and peripapillary regions of patients with Generalized Anxiety Disorder (GAD) using optical coherence tomography (OCT) and OCT angiography (OCT-A). We compared these changes with healthy controls to determine GAD’s impact on the retina. Methods An observational case-control study was conducted from January to May 2024 at the Benghazi Teaching Eye Hospital, including 40 GAD patients and 40 healthy controls aged 30–65 years. Participants underwent comprehensive ophthalmic assessments, including OCT and OCT-A. We analyzed retinal parameters such as central macular thickness (CMT), Macular volume, Ganglion cell layer thickness, and retinal nerve fiber layer (RNFL) thickness, along with vessel density in the macular and peripapillary regions. Results GAD patients exhibited significantly lower CMT (243.30±21.15 μm vs. 268.79±17.34 μm, P=0.001), Macular volume (9.48±0.62 mm3 vs. 10.17±0.39 mm3, P=0.001), Ganglion cell layer thickness (83.60±78.24 μm vs. 92.30±74.73 μm, P=0.001), and total RNFL thickness (93.90±11.05 μm vs. 97.76±8.67 μm, P=0.001) compared with controls. RNFL thinning was noted in the nasal, inferior, and temporal regions. However, OCT-A revealed no significant differences in vessel density in the peripapillary and central macular areas between GAD patients and controls. Conclusion Patients with GAD exhibit significant retinal structural changes, particularly in the macular and RNFL regions. However, no significant differences in retinal vessel density were observed. These findings suggest that GAD may impact retinal morphology but not vascular density, indicating potential biomarkers for early detection and monitoring of GAD-related neurodegeneration.

Superficial vessel density and determinants of macular microvasculature in healthy Chinese subjects

BackgroundThe purpose of this study was to investigate the vessel density (VD) of the macular superficial capillary plexus (SCP) and its associated factors in healthy subjects. MethodsThis prospective, cross-sectional study enrolled healthy Chinese volunteers. The optical coherence tomography angiography (OCTA) was used to measure the superficial VD in macula. A generalized estimation equation (GEE) model was used to analyze the ocular and systemic associated factors. ResultsA total of 516 eyes of 262 healthy subjects were included (mean age 38.59 ± 21.03 years). The total VD of macular SCP was 16.85 ± 2.28 mm−1. The VD in the inner ring and outer ring were significantly higher than that in the central ring, with the density being highest in nasal quadrant and lowest in the superior quadrant. After adjusting the ocular and systemic factors, age (β=-0.0085, P = 0.0122) and SSI (β=1.3261, P < 0.001) were significantly associated with total VD of the macular SCP. ConclusionsAmong the healthy Chinese subjects included in the study, the mean total VD of macular SCP was 16.85 ± 2.28mm-1. The macular superficial capillary density was positively associated with age and SSI. Further studies are still required to better understand the change of macular VD in aging and other pathological conditions.

The Potential Utility of Retinal Biomarkers to Index Central Nervous System Effects of Repetitive Blast Exposure in Military Personnel.

Military personnel exposed to blasts receive repeated subconcussive head impacts. Although these events typically do not cause immediate symptoms and do not lead to medical evaluation, the cumulative effects of subconcussive impacts can be significant and can include postconcussive symptoms, changes in brain structure and function, long-term cognitive dysfunction, depression, and chronic traumatic encephalopathy. Retinal measures such as thickness of retinal neural layers, density of retinal microvasculature, and strength of retinal neuronal firing are associated with cognitive function and brain structure and function in healthy populations and in neurodegenerative disease cohorts, and changes over time in retinal indices predict cognitive decline and brain atrophy in longitudinal studies in a range of medical populations. This commentary highlights the potential benefits of using retinal biomarkers in the routine screening and monitoring of brain health in warfighters and veterans. Retinal measures can be rapidly acquired (often in a matter of seconds) using methods that are inexpensive and noninvasive, and they can be collected with movable and often portable equipment that uses automated scoring routines that can be used for prediction and decision-making at the individual level. To date, however, retinal biomarkers have not been included in studies of blast overpressure exposures in military personnel. Despite this, preclinical and human evidence suggests that they could be among the most effective methods for tracking central nervous system damage in people exposed to repeated blasts. Retinal biomarkers could also contribute to brief test batteries to determine who is most at risk for long-term negative effects of future exposures. In addition, the sensitivity of retinal indices to blast exposure and mild traumatic brain injury suggests that they should be incorporated into research on strategies to minimize or prevent blast-related short- and long-term central nervous system changes in blast-exposed military personnel.

Electroacupuncture facilitates vascular normalization by inhibiting Glyoxalase1 in endothelial cells to attenuate glycolysis and angiogenesis in triple-negative breast cancer

Recent therapeutic strategies for the treatment of triple-negative breast cancer (TNBC) have shifted the focus from vascular growth factors to endothelial cell metabolism. This study highlights the underexplored therapeutic potential of peri-tumoral electroacupuncture, a globally accepted non-pharmacological intervention for TNBC, and molecular mechanisms. Our study showed that peri-tumoral electroacupuncture effectively reduced the density of microvasculature and enhanced vascular functionality in 4T1 breast cancer xenografts, with optimal effects on day 3 post-acupuncture. The timely integration of peri-tumoral electroacupuncture amplified the anti-tumor efficacy of paclitaxel. Multi-omics analysis revealed Glyoxalase 1 (Glo1) and the associated methylglyoxal-glycolytic pathway as key mediators of electroacupuncture-induced vascular normalization. Peri-tumoral electroacupuncture notably reduced Glo1 expression in the endothelial cells of 4T1 xenografts. Using an in vivo matrigel plug angiogenesis assay, we demonstrated that either Glo1 knockdown or electroacupuncture inhibited angiogenesis. In contrast, Glo1 overexpression increased blood vessel formation. In vitro pharmacological inhibition and genetic knockdown of Glo1 in human umbilical vein endothelial cells inhibited proliferation and promoted apoptosis via downregulating the methylglyoxal-glycolytic pathway. The study using the Glo1-silenced zebrafish model further supported the role of Glo1 in vascular development. This study underscores the pivotal role of Glo1 in peri-tumoral electroacupuncture, spotlighting a promising avenue for enhancing vascular normalization and improving TNBC treatment outcomes.

Non-invasive optoacoustic imaging of dermal microcirculatory revascularization in diet-induced obese mice undergoing exercise intervention

Microcirculatory dysfunction has been observed in the dermal white adipose tissue (dWAT) and subcutaneous white adipose tissue (scWAT) of obese humans and has been proposed as an early prediction marker for cardio-metabolic disease progression. In-vivo visualization and longitudinal monitoring of microvascular remodeling in these tissues remains challenging. We compare the performance of two optoacoustic imaging methods, i.e. multi-spectral optoacoustic tomography (MSOT) and raster-scanning optoacoustic mesoscopy (RSOM) in visualizing lipid and hemoglobin contrast in scWAT and dWAT in a mouse model of diet-induced obesity (DIO) undergoing voluntary wheel running intervention for 32 weeks. MSOT visualized lipid and hemoglobin contrast in murine fat depots in a quantitative manner even at early stages of DIO. We show for the first time to our knowledge that RSOM allows precise visualization of the dWAT microvasculature and provides quantitative readouts of skin layer thickness and vascular density in dWAT and dermis. Combination of MSOT and RSOM resolved exercise-induced morphological changes in microvasculature density, tissue oxygen saturation, lipid and blood volume content in dWAT and scWAT. The combination of MSOT and RSOM may allow precise monitoring of microcirculatory dysfunction and intervention response in dWAT and scWAT in a mouse model for DIO. Our findings have laid out the foundation for future clinical studies using optoacoustic-derived vascular readouts from adipose tissues as a biomarker for monitoring microcirculatory function in metabolic disease.

Quantitative Measurement and Analysis of the Radial Peripapillary and Central Macular Microvasculature Density in Early Open-Angle Glaucoma Using OCT Angiography

Background Glaucoma is a prevalent cause of visual impairment, and early detection is critical to prevent progressive vision loss. Optical coherence tomography angiography (OCT-A) enables visualization of retinal and optic nerve microvasculature and holds promise for early glaucoma detection. Aims This study aims to assess microvasculature density alterations in glaucoma using OCT-A. Methods An observational cross-sectional case–control study was conducted at Benghazi Eye Hospital in Libya from December 2022 to August 2023. It included 160 eyes of 126 patients diagnosed with primary open-angle glaucoma and 120 eyes of 96 control subjects. OCT-A was performed using the Swept-Source OCT DRI Triton. Vessel density measurements were obtained from OCT-A images using new built-in software within the device. The data presented as mean ± standard deviation and percentages were analyzed using SPSS. The statistical significance of the differences between different groups was determined using independent t-tests, and the level of significance was set at p-value less than 0.05. Results OCT-A demonstrated significantly reduced microvasculature density in glaucoma eyes across all optic nerve sectors including the inferior (29.0%), superior (25.2%), central (23.5%), nasal (9.9%), and temporal sectors (9.3%; p &lt; 0.02). Moreover, glaucoma patients exhibited a significant reduction in vessel density in the superior macula (17% reduction), the temporal (15.7%,), nasal (12.9%), and inferior sectors (12.6%) (p &lt; 0.002). No statistical difference was seen with vessel density of the central sector (0.49%, p &gt;0.05). Conclusion OCT-A demonstrated significant reductions in microvasculature density in glaucoma patients compared with healthy controls. These findings support the association between glaucoma and vascular changes. Additionally, the TOPCON Swept-Source OCT DRI Triton new software shows promise as a valuable tool for early detection and monitoring of glaucoma-related vascular changes.

Automated evaluation of parapapillary choroidal microvasculature in crowded optic discs: a controlled, optical coherence tomography angiography study.

To compare superficial and deep vascular properties of optic discs between crowded discs and controls using optical coherence tomography angiography (OCT-A). Thirty patients with crowded discs, and 47 control subjects were enrolled in the study. One eye of each individual was included and OCT-A scans of optic discs were obtained in a 4.5×4.5 mm2 rectangular area. Radial peripapillary capillary (RPC) density, peripapillary retinal nerve fiber layer (pRNFL) thickness, cup volume, rim area, disc area, cup-to-disc (c/d) area ratio, and vertical c/d ratio were obtained automatically using device software. Automated parapapillary choroidal microvasculature (PPCMv) density was calculated using MATLAB software. When the vertical c/d ratio of the optic disc was absent or small cup, it was considered as a crowded disc. The mean signal strength index of OCT-A images was similar between the crowded discs and control eyes (P=0.740). There was no difference in pRNFL between the two groups (P=0.102). There were no differences in RPC density in whole image (P=0.826) and peripapillary region (P=0.923), but inside disc RPC density was higher in crowded optic discs (P=0.003). The PPCMv density in the inner-hemisuperior region was also lower in crowded discs (P=0.026). The pRNFL thickness was positively correlated with peripapillary RPC density (r=0.498, P<0.001). The inside disc RPC density was negatively correlated with c/d area ratio (r=-0.341, P=0.002). The higher inside disc RPC density and lower inner-hemisuperior PPCMv density are found in eyes with crowded optic discs.

The Microvascular Network Density and Morphology Affect the Radiotherapy Outcome at the Microscale Level: A Computational Analysis.

Half of the cancer patients are treated with radiotherapy to obtain tumor control while minimizing radiation toxicity. In this context, hypoxia is known to determine treatment resistance, driving tumor relapse after treatment. Therefore, we investigated the role of microvasculature density and morphology in shaping the tissue oxygen distribution and consequently affecting the treatment outcome. We developed an advanced computational model to describe oxygen delivery in the vascular network and the surrounding tissue. A peculiar aspect of the model is its mesoscale mixed-dimensional approach, which allows the explicit inclusion of vascular network geometry and the description of the red blood cells' effect. The oxygen delivery is modeled considering both diffusive and convective phenomena. A 30 × 2GyRBE treatment is delivered in silico simulating photons, protons, and carbon ions. First, we estimated the surviving fraction by the classical linear quadratic model modified to account for the oxygen effect. Then, leveraging the 3D description of the surviving fraction at the microenvironment scale, we compute the local tumor control probability (LTCP) in different oxygenation states (reference, acute hypoxia, high oxygen consumption). We report correlations between the LTCP and the hypoxic volume fractions (with pO2 lower than 1 mmHg) starting with photons. These hypoxic regions are present locally, even in highly vascularized tissue, if the network is not uniformly distributed, as it might be in cancer. They are also present in tissue with low microvascular density, even with regular morphology. Interestingly, the domain considered is comparable to or smaller than the clinical imaging standard voxel dimension, and the average oxygen partial pressure in the tissue region fails to spot treatments with low LTCP, questioning whether these hypoxia areas are visible clinically via imaging. Protons have a similar effect, highlighting a similar behavior across the oxygenation levels at the microscale level. Finally, carbon ions seem more effective than photons and protons in the presence of hypoxia due to the lower oxygen effect at high LET. For this reason, the treatment with carbon ions results in high LTCPs, whatever vascular network is considered (density and morphology differences). These results show the effect of microvascular density and regularity on the radiotherapy outcome and they help us understand how the microvascular network morphology affects tumor oxygenation and the radiotherapy outcome.

Opening a window to skin biomarkers for diabetes stage with optoacoustic mesoscopy

Being the largest and most accessible organ of the human body, the skin could offer a window to diabetes-related complications on the microvasculature. However, skin microvasculature is typically assessed by histological analysis, which is not suited for applications to large populations or longitudinal studies. We introduce ultra-wideband raster-scan optoacoustic mesoscopy (RSOM) for precise, non-invasive assessment of diabetes-related changes in the dermal microvasculature and skin micro-anatomy, resolved with unprecedented sensitivity and detail without the need for contrast agents. Providing unique imaging contrast, we explored a possible role for RSOM as an investigational tool in diabetes healthcare and offer the first comprehensive study investigating the relationship between different diabetes complications and microvascular features in vivo. We applied RSOM to scan the pretibial area of 95 participants with diabetes mellitus and 48 age-matched volunteers without diabetes, grouped according to disease complications, and extracted six label-free optoacoustic biomarkers of human skin, including dermal microvasculature density and epidermal parameters, based on a novel image-processing pipeline. We then correlated these biomarkers to disease severity and found statistically significant effects on microvasculature parameters as a function of diabetes complications. We discuss how label-free RSOM biomarkers can lead to a quantitative assessment of the systemic effects of diabetes and its complications, complementing the qualitative assessment allowed by current clinical metrics, possibly leading to a precise scoring system that captures the gradual evolution of the disease.

Invited Session V: The eye as a window to systemic and neurodegenerative health: Disturbances of retinal structure in schizophrenia spectrum disorders and their clinical implications.

Schizophrenia (SZ) is a disabling psychiatric condition that is characterized by disturbances in reality testing (e.g., hallucinations, delusions) during its acute phase and by many brain-related impairments. It has become apparent, however, that the CNS disturbance in SZ extends to the retina. Studies have demonstrated retinal structural (neural and vascular) and functional (e.g., ERG) changes in SZ, although findings vary in effect size and confounds are often present. This presentation will review several studies from my lab that used OCT and OCTA to investigate retinal characteristics of people with SZ, with a focus on characterizing within-group heterogeneity. Results indicate that: 1) retinal thinning is present at the macula, but not until several years after the first psychotic episode; 2) reductions in microvasculature density are observable as early as the first episode and worsen over time at a greater rate than expected by age alone; 3) microvasculature reductions are present in both the superficial and deep retinal layers; and 4) issues such as diabetes, smoking, and obesity must be carefully considered when studying the retina in SZ. Findings will be discussed within the context of the emerging view that SZ is multi-system disorder with a significant vascular-ischemic component.

Automated evaluation of parapapillary choroidal microvasculature in thyroid eye disease.

The purpose of the study was to investigate the parapapillary choroidal microvasculature in thyroid eye disease (TED) using optical coherence tomography angiography (OCTA). Only one eye of each subject was included in the study. Patients with TED and controls were included in the study. Participants were divided into three groups: control, inactive TED (ITED) and active TED (ATED). OCTA scans of the optic discs were obtained in a 4.5 × 4.5-mm rectangular area. Radial peripapillary capillary (RPC) density and peripapillary retinal nerve fibre layer (pRNFL) thickness were automatically calculated by the device software. Parapapillary choroidal microvasculature (PPCMv) density was automatically calculated using MATLAB software. Forty-one patients with TED and 40 controls were included in the study. RPC density was significantly decreased in the ATED and dysthyroid optic neuropathy (DON) group compared to the controls and ITED group. There was significant increase in pRNFL in the ATED group. PPCMv density increased in the ATED group compared to the controls in whole ring area. The RPC density was significantly correlated with the TSHr Ab level (r < -0.396, p < 0.001). Clinical activity score correlated positively with PPCMv density (r = 0.349, p = 0.001) but negatively with RPC density (r = -0.321, p = 0.004). Changes in peripapillary microvascular perfusion may play a role in the development of DON. As the severity of TED increases with clinical activity, so do the changes observed in peripapillary parameters. The decrease in RPC density may be due to compression caused by optic disc oedema, which may result in reduced blood flow. The increase in PPCMv density may be related to factors such as orbital congestion.

Association of retinal microvascular density and complexity with incident coronary heart disease

Background and aimsThe high mortality rate and huge disease burden of coronary heart disease (CHD) highlight the importance of its early detection and timely intervention. Given the non-invasive nature of fundus photography and recent development in the quantification of retinal microvascular parameters with deep learning techniques, our study aims to investigate the association between incident CHD and retinal microvascular parameters. MethodsUK Biobanks participants with gradable fundus images and without a history of diagnosed CHD at recruitment were included for analysis. A fully automated artificial intelligence system was used to extract quantitative measurements that represent the density and complexity of the retinal microvasculature, including fractal dimension (Df), number of vascular segments (NS), vascular skeleton density (VSD) and vascular area density (VAD). ResultsA total of 57,947 participants (mean age 55.6 ± 8.1 years; 56% female) without a history of diagnosed CHD were included. During a median follow-up of 11.0 (interquartile range, 10.88 to 11.19) years, 3211 incident CHD events occurred. In multivariable Cox proportional hazards models, we found decreasing Df (adjusted HR = 0.80, 95% CI, 0.65–0.98, p = 0.033), lower NS of arteries (adjusted HR = 0.69, 95% CI, 0.54–0.88, p = 0.002) and venules (adjusted HR = 0.77, 95% CI, 0.61–0.97, p = 0.024), and reduced arterial VSD (adjusted HR = 0.72, 95% CI, 0.57–0.91, p = 0.007) and venous VSD (adjusted HR = 0.78, 95% CI, 0.62–0.98, p = 0.034) were related to an increased risk of incident CHD. ConclusionsOur study revealed a significant association between retinal microvascular parameters and incident CHD. As the lower complexity and density of the retinal vascular network may indicate an increased risk of incident CHD, this may empower its prediction with the quantitative measurements of retinal structure.

Microcirculation surrounding end-stage human chronic skin wounds is associated with endoglin/CD146/ALK-1 expression, endothelial cell proliferation and an absence of p16Ink4a.

Angiogenesis is an essential part of normal skin healing, re-establishing blood flow in developing granulation tissue. Non-healing skin wounds are associated with impaired angiogenesis and although the role of re-establishing macroscopic blood flow to limbs to prevent wound chronicity is well investigated, less is known about vascular alterations at the microcirculatory level. We hypothesised that significant phenotypic changes would be evident in blood vessels surrounding chronic skin wounds. Wound edge tissue, proximal to wound (2 cm from wound edge) and non-involved skin (>10 cm from wound edge) was harvested under informed consent from 20 patients undergoing elective amputation due to critical limb ischemia. To assess blood vessel structure and viability, tissue was prepared for histological analysis and labelled with antibodies specific for PECAM-1 (CD31), CD146, endoglin, ALK-1, ALK-5, and p16Ink4a as a marker of cellular senescence. Density of microvasculature was significantly increased in wound edge dermis, which was concomitant with increased labelling for endoglin and CD146. The number of CD31 positive vessel density was unchanged in wound edge tissue relative to non-involved tissue. Co-labelling of endoglin with the transforming growth factor receptor ALK-1, and to a lesser extent ALK-5, demonstrated activation of endothelial cells which correlated with PCNA labelling indicative of proliferation. Analysis of p16Ink4a staining showed a complete lack of immunoreactivity in the vasculature and dermis, although staining was evident in sub-populations of keratinocytes. We conclude that the endoglin-ALK-1-endothelial proliferation axis is active in the vasculature at the edge of chronic skin wounds and is not associated with p16Ink4a mediated senescence. This information could be further used to guide treatment of chronic skin wounds and optimise debridement protocols.

The longevity-associated BPIFB4 gene supports cardiac function and vascularization in ageing cardiomyopathy.

The ageing heart naturally incurs a progressive decline in function and perfusion that available treatments cannot halt. However, some exceptional individuals maintain good health until the very late stage of their life due to favourable gene-environment interaction. We have previously shown that carriers of a longevity-associated variant (LAV) of the BPIFB4 gene enjoy prolonged health spans and lesser cardiovascular complications. Moreover, supplementation of LAV-BPIFB4 via an adeno-associated viral vector improves cardiovascular performance in limb ischaemia, atherosclerosis, and diabetes models. Here, we asked whether the LAV-BPIFB4 gene could address the unmet therapeutic need to delay the heart's spontaneous ageing. Immunohistological studies showed a remarkable reduction in vessel coverage by pericytes in failing hearts explanted from elderly patients. This defect was attenuated in patients carrying the homozygous LAV-BPIFB4 genotype. Moreover, pericytes isolated from older hearts showed low levels of BPIFB4, depressed pro-angiogenic activity, and loss of ribosome biogenesis. LAV-BPIFB4 supplementation restored pericyte function and pericyte-endothelial cell interactions through a mechanism involving the nucleolar protein nucleolin. Conversely, BPIFB4 silencing in normal pericytes mimed the heart failure pericytes. Finally, gene therapy with LAV-BPIFB4 prevented cardiac deterioration in middle-aged mice and rescued cardiac function and myocardial perfusion in older mice by improving microvasculature density and pericyte coverage. We report the success of the LAV-BPIFB4 gene/protein in improving homeostatic processes in the heart's ageing. These findings open to using LAV-BPIFB4 to reverse the decline of heart performance in older people.

Detection of Stroke with Retinal Microvascular Density and Self-Supervised Learning Using OCT-A and Fundus Imaging.

Acute cerebral stroke is a leading cause of disability and death, which could be reduced with a prompt diagnosis during patient transportation to the hospital. A portable retina imaging system could enable this by measuring vascular information and blood perfusion in the retina and, due to the homology between retinal and cerebral vessels, infer if a cerebral stroke is underway. However, the feasibility of this strategy, the imaging features, and retina imaging modalities to do this are not clear. In this work, we show initial evidence of the feasibility of this approach by training machine learning models using feature engineering and self-supervised learning retina features extracted from OCT-A and fundus images to classify controls and acute stroke patients. Models based on macular microvasculature density features achieved an area under the receiver operating characteristic curve (AUC) of 0.87-0.88. Self-supervised deep learning models were able to generate features resulting in AUCs ranging from 0.66 to 0.81. While further work is needed for the final proof for a diagnostic system, these results indicate that microvasculature density features from OCT-A images have the potential to be used to diagnose acute cerebral stroke from the retina.

Combination of indirect revascularization and endothelial progenitor cell transplantation improved cerebral perfusion and ameliorated tauopathy in a rat model of bilateral ICA ligation

ObjectiveEndothelial progenitor cells (EPCs) contribute to the recovery of neurological function after ischemic stroke. Indirect revascularization has exhibited promising effects in the treatment of cerebral ischemia related to moyamoya disease and intracranial atherosclerotic disease. The role of EPCs in augmenting the revascularization effect is not clear. In this study, we investigated the therapeutic effects of indirect revascularization combined with EPC transplantation in rats with chronic cerebral ischemia.MethodsChronic cerebral ischemia was induced by bilateral internal carotid artery ligation (BICAL) in rats, and indirect revascularization by encephalo-myo-synangiosis (EMS) was performed 1 week later. During the EMS procedure, intramuscular injection of EPCs and the addition of stromal cell-derived factor 1 (SDF-1), and AMD3100, an SDF-1 inhibitor, were undertaken, respectively, to investigate their effects on indirect revascularization. Two weeks later, the cortical microcirculation, neuronal damage, and functional outcome were evaluated according to the microvasculature density and partial pressure of brain tissue oxygen (PbtO2), regional blood flow, expression of phosphorylated Tau (pTau), TUNEL staining and the rotarod performance test, respectively.ResultsThe cortical microcirculation, according to PbtO2 and regional blood flow, was impaired 3 weeks after BICAL. These impairments were improved by the EMS procedure. The regional blood flow was further increased by the addition of SDF-1 and decreased by the addition of AMD3100. Intramuscular injection of EPCs further increased the regional blood flow as compared with the EMS group. The rotarod test results showed that the functional outcome was best in the EMS combined with EPC injection group. Western blot analysis showed that the EMS combined with EPC treatment group had significantly decreased expressions of phosphorylated Tau and phosphorylated glycogen synthase kinase 3 beta (Y216 of GSK-3β). pTau and TUNEL-positive cells were markedly increased at 3 weeks after BICAL induction. Furthermore, the groups treated with EMS combined with SDF-1 or EPCs exhibited marked decreases in the pTau expression and TUNEL-positive cells, whereas AMD3100 treatment increased TUNEL-positive cells.ConclusionThe results of this study suggested that indirect revascularization ameliorated the cerebral ischemic changes. EPCs played a key role in augmenting the effect of indirect revascularization in the treatment of chronic cerebral ischemia.

DWI Metrics Differentiating Benign Intraductal Papillary Mucinous Neoplasms from Invasive Pancreatic Cancer: A Study in GEM Models.

Simple SummaryIntraductal papillary mucinous neoplasms (IPMN) are common premalignant precursors to pancreatic ductal adenocarcinoma (PDAC). However, current guidelines for clinical management of IPMN are suboptimal and limited to information obtained through anatomical imaging and cytology of biopsy samples. Our results have suggested that diffusion-weighted MRI, an imaging method sensitive to tumor cell density, architecture, and microenvironment is able to differentiate mouse models of IPMN versus PDAC.KPC (KrasG12D:Trp53R172H:Pdx1-Cre) and CKS (KrasG12D:Smad4L/L:Ptf1a-Cre) mice are genetically engineered mouse (GEM) models that capture features of human pancreatic ductal adenocarcinoma (PDAC) and intraductal papillary mucinous neoplasms (IPMN), respectively. We compared these autochthonous tumors using quantitative imaging metrics from diffusion-weighted MRI (DW-MRI) and dynamic contrast enhanced (DCE)-MRI in reference to quantitative histological metrics including cell density, fibrosis, and microvasculature density. Our results revealed distinct DW-MRI metrics between the KPC vs. CKS model (mimicking human PDAC vs. IPMN lesion): the apparent diffusion coefficient (ADC) of CKS tumors is significantly higher than that of KPC, with little overlap (mean ± SD vs. , ) despite intratumor and intertumor variability. Kurtosis index (KI) is also distinctively separated in the two models. DW imaging metrics are consistent with growth pattern, cell density, and the cystic nature of the CKS tumors. Coregistration of ex vivo ADC maps with H&E-stained sections allowed for regional comparison and showed a correlation between local cell density and ADC value. In conclusion, studies in GEM models demonstrate the potential utility of diffusion-weighted MRI metrics for distinguishing pancreatic cancer from benign pancreatic cysts such as IPMN.

Effects of Smoking on Optic Nerve Head Microvasculature Density in Glaucoma.

Decreased superficial whole image capillary density was observed in open angle glaucoma (OAG) patients with high smoking intensity. To investigate the effects of smoking on optic nerve head capillary density measured by optical coherence tomography angiography in patients with OAG. In this retrospective cross-sectional study, perimetric and preperimetric glaucoma patients enrolled in the Diagnostic Innovations in Glaucoma Study (DIGS) with optical coherence tomography angiography follow-up were included. Univariable and multivariable linear mixed analysis were performed to determine the effects of different variables on the superficial whole image capillary density. A total of 432 eyes of 271 glaucoma patients comprising 63 preperimetric (106 eyes) and 208 perimetric OAG (326 eyes) were included. A history of tobacco consumption was reported in 105 (38.8%). Among smokers, mean (95% confidence interval) smoking intensity was 12.8 (10.2, 15.5) pack-years. After adjusting for age, glaucoma severity and other confounders, each 10 pack-year increase in smoking intensity (95% confidence interval) was associated with -0.54 (-1.06, -0.02) % lower whole image capillary density ( P =0.041). Smoking intensity is associated with reduced optic nerve vessel density in glaucoma.

Angiogenic gene characterization and vessel permeability of dermal microvascular endothelial cells isolated from burn hypertrophic scar

Hypertrophic scar (HTS) formation is a common challenge for patients after burn injury. Dermal microvascular endothelial cells (DMVECs) are an understudied cell type in HTS. An increase in angiogenesis and microvessel density can be observed in HTS. Endothelial dysfunction may play a role in scar development. This study aims to generate a functional and expression profile of HTS DMVECs. We hypothesize that transcript and protein-level responses in HTS DMVECs differ from those in normal skin (NS). HTSs were created in red Duroc pigs. DMVECs were isolated using magnetic-activated cell sorting with ulex europaeus agglutinin 1 (UEA-1) lectin. Separate transwell inserts were used to form monolayers of HTS DMVECs and NS DMVECs. Cell injury was induced and permeability was assessed. Gene expression in HTS DMVECS versus NS DMVECs was measured. Select differentially expressed genes were further investigated. HTS had an increased area density of dermal microvasculature compared to NS. HTS DMVECs were 17.59% less permeable than normal DMVECs (p < 0.05). After injury, NS DMVECs were 28.4% and HTS DMVECs were 18.8% more permeable than uninjured controls (28.4 ± 4.8 vs 18.8 ± 2.8; p = 0.11). PCR array identified 31 differentially expressed genes between HTS and NS DMVECs, of which 10 were upregulated and 21 were downregulated. qRT-PCR and ELISA studies were in accordance with the array. DMVECs expressed a mixed profile of factors that can contribute to and inhibit scar formation. HTS DMVECs have both a discordant response to cellular insults and baseline differences in function, supporting their proposed role in scar pathology. Further investigation of DMVECs is warranted to elucidate their contribution to HTS pathogenesis.