The ocular microcirculation represents an important target to treat inflammatory diseases of eye, where impairment of microvascular blood flow plays key role as, for example, in anterior uveitis. To evaluate novel interventions targeting the microcirculation, appropriate and reliable tools to study this particular microvascular bed are needed. Intravital microscopy (IVM) belongs to several methods allowing evaluation of microcirculation experimentally, even in small animals. The aim of our study was to examine the iridial microcirculation (IMIC) in uveitis induced by local or systemic endotoxin administration in rats and mice by IVM and to propose new parameters to quantify the changes within the IMIC. Systemic inflammation was induced in rats by intravenous endotoxin administration, control group received normal saline intravenously. Local inflammation was induced in mice by intravitreal endotoxin administration, the control group received normal saline intravitreally. IVM of IMIC was performed in animals receiving systemic endotoxin prior injection and 1 and 2 h afterwards, respectively, in animals receiving intravitreal endotoxin/saline prior local injection and 5 h afterwards. Obtained video recordings were analyzed off-line. Functional capillary density (FCD) and dysfunctional capillary density (DCD) were evaluated for description of IMIC, and calculation of FCD/DCD ratio was performed. In systemic inflammation, FCD was significantly decreased compared to control animals. In local inflammation, the number of functional capillaries in the IMIC was significantly reduced following the endotoxin challenge. Analysis of the DCD revealed a significant increase in capillaries with reduced perfusion after intravitreal endotoxin administration and right shift of the FCD/DCD ratio was observed after endotoxin local injection. Detecting and quantifying changes in IMIC during systemic or local inflammation in experimental animals by IVM was feasible. Therefore, IVM of the IMIC represents a valuable tool to evaluate and quantify inflammatory changes in experimental eye disease.
- MeSH
- endotoxiny aplikace a dávkování toxicita MeSH
- intravitální mikroskopie metody MeSH
- kapiláry patologie MeSH
- krysa rodu rattus MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- uveitida chemicky indukované patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
For microcirculation research there is a need for baseline data and feasibility protocols describing microcirculation of various organs. The aim of our study was to examine the reliability and reproducibility of sidestream dark-field (SDF) imaging within the renal cortical microcirculation in rats. Renal microcirculation was observed using SDF probe placed on the exposed renal surface via the upper midline laparotomy. Video sequences recorded intermittently in short apneic pauses were analyzed off-line by using AVA 3.0 software (MicroVision Medical, Amsterdam, the Netherlands). Results are expressed as mean (SD) or median (25-75% percentiles). We obtained 60 clear sequences from all recorded analyzable videos from all the animals. The total small vessel and all vessel density (in mm.mm(-2) ) were (28.79 ± 0.40) and (28.95 ± 0.40), respectively. The perfused small and all vessel density were (28.79 ± 0.40) and (28.95 ± 0.40), respectively. The DeBacker Score was (19.14 ± 0.43), the proportion of perfused vessels was 100% (100-100%) and the microvascular flow index was 3.49 (3-3.75). We conclude SDF imaging provides a reliable method to examine the renal microvascular bed in vivo and thus can be used for the study of the renal cortical vascular network in various experimental diseases models and clinical settings.
- MeSH
- krysa rodu rattus MeSH
- ledviny anatomie a histologie fyziologie MeSH
- mikrocirkulace * MeSH
- videomikroskopie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Nizozemsko MeSH
BACKGROUND: Pregnancy places significant demands on the cardiovascular system leading to measurable changes in the macrocirculation and potentially the microcirculation. During labour, both uterine contractions and labour pain can further impact cardiovascular status. The objective of this observational study was to compare sublingual microcirculation in labouring parturients before and after epidural analgesia. METHODS: Healthy pregnant, labouring women requesting epidural analgesia were approached to participate. Participants with cardiovascular disease, diabetes, obesity, smoking or caffeine intake were excluded. The sidestream dark field device was applied to the sublingual mucosa obtaining images of at least 20 seconds in 5 visual fields before and after epidural analgesia. Video clips were analyzed randomly and blindly. The primary outcome was mean microvascular flow index (MFI). RESULTS: Twelve participants completed this study. The results demonstrate no statistically significant difference in the MFI during labour pain (2.9±0.1) compared to after epidural analgesia (3.0±0.04, p = 0.31). Furthermore, there were no statistically significant differences in any secondary outcomes. CONCLUSION: Our findings indicate that epidural analgesia may not impact sublingual microcirculation in labouring women. This agrees with literature supporting epidural analgesia as a safe, appropriate method of pain relief during labour with limited impact on peripheral macro or microcirculation.
- MeSH
- analgézie porodnická metody MeSH
- diagnostické zobrazování metody MeSH
- dospělí MeSH
- lidé MeSH
- mikrocirkulace MeSH
- porodní bolesti farmakoterapie MeSH
- prospektivní studie MeSH
- těhotenství MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
BACKGROUND: The microcirculation is responsible for distribution of blood within tissues, delivery of oxygen and other nutrients, and regulation of blood pressure. The objective of this study was to compare the sublingual microcirculation of pregnant participants to that of comparable non-pregnant volunteers. METHODS: Two groups of participants were recruited: a group of pregnant, non-laboring women with singleton pregnancies at term gestation and a control group of age-comparable non-pregnant volunteers. A sidestream dark field imaging device was applied to the sublingual mucosal surface obtaining a steady image for at least 20 s duration, in five visual fields. The resultant five video clips per participant were analyzed blindly and at random to prevent coupling between images. The mean microvascular flow index values for each group were compared using a paired t-test. RESULTS: Thirty-seven participants were recruited (19 pregnant, 18 non-pregnant); a single pregnant participant was withdrawn because of technical issues. Baseline characteristics were similar with the exception of weight and body mass index. The mean microvascular flow index was significantly higher in the pregnant group 2.7 ± 0.2 compared to the non-pregnant group 2.5 ± 0.3 (P = 0.021), while the perfused vessel density and proportion of perfused vessels were not significantly different (P = 0.707 and 0.403, respectively). CONCLUSION: The microvascular flow index of pregnant women is higher than a comparable non-pregnant group, which appears to correlate with the physiological changes of pregnancy.
- MeSH
- audiovizuální záznam metody MeSH
- dospělí MeSH
- lidé MeSH
- mikrocirkulace fyziologie MeSH
- prospektivní studie MeSH
- těhotenství MeSH
- ústní spodina krevní zásobení MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
- práce podpořená grantem MeSH
The microcirculation, like all physiological systems undergoes modifications during the course of pregnancy. These changes aid the adaption to the new anatomical and physiological environment of pregnancy and ensure adequate oxygen supply to the fetus. Even though the microcirculation is believed to be involved in major pregnancy related pathologies, it remains poorly understood. The availability of safe and non-interventional technologies enabling scientists to study the intact microcirculation of the pregnant patient will hopefully expand our understanding. In this article we review the physiological changes occurring in the microcirculation during pregnancy and the role of the microcirculation in gestational related pathologies. We will also describe the available techniques for the measurement and evaluation of the microcirculation. Lastly we will highlight the possible fields in which these techniques could be utilized to help provide a clearer view of the microcirculation in the pregnant woman.
- MeSH
- dospělí MeSH
- komplikace těhotenství patofyziologie MeSH
- lidé MeSH
- mikrocirkulace fyziologie MeSH
- těhotenství fyziologie MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství fyziologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Sidestream dark field imaging represents a novel, noninvasive method to study the microcirculation in humans and animals. To-date, it has been used extensively in various peripheral tissues (e.g. sublingual area, intestinal mucosa), however no data for the ocular vasculature, including the iridial microcirculation, are currently available. Therefore, the aim of this study was to examine the reliability and reproducibility of sidestream dark field imaging within the iridial microcirculation in experimental animals. Male Lewis rats were anaesthetized and the iris microvasculature was observed using an sidestream dark field probe gently placed against a cover slip covering the right eye. All video sequences recorded were analysed off-line by using AVA 3.0 software (MicroVision Medical, Amsterdam, The Netherlands). Results are expressed as mean (±SE) or median (interquartile range). Clear images were recorded from each animal and the total number of analysable video sequences was 50. All raw data for selected vessel density parameters passed normality test. The total all and small vessel density (in mm mm(-2) ) were 22,6 (±0,58) and 19,6 (±0,68), respectively. The perfused all and small vessel density were 20,9 (±0,61) and 19,1 (±0,65), respectively. The mean values of all iris vessel density parameters are shown in Figure 4. The DeBacker Score (n/mm) was 15,2 (±0,45), the proportion of perfused vessel was 94,5% (89,8-99,1%), and the MFI was 3 points (3-3). Taken together, these results indicate that SDF imaging provides a reliable and noninvasive method to examine the iridial microvascular bed in vivo and, thus, may provide unique opportunities for the study of the iridial vascular network in various experimental and clinical settings and disease models.
- MeSH
- iris anatomie a histologie fyziologie MeSH
- krysa rodu rattus MeSH
- mikrocévy anatomie a histologie fyziologie MeSH
- počítačové zpracování obrazu metody MeSH
- videomikroskopie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Disturbance of capillary perfusions due to leukocyte adhesion, disseminated intravascular coagulation, tissue edema is critical components in the pathophysiology of sepsis. Alterations in brain microcirculation during sepsis are not clearly understood. The aim of this study is to gain an improved understanding of alterations through direct visualization of brain microcirculations in an experimental endotoxemia using intravital microscopy (IVM). Endotoxemia was induced in Lewis rats with Lipopolysaccharide (LPS, 15 mg/kg i.v.). The dura mater was removed via a cranial window to expose the pial vessels on the brain surface. Using fluorescence dyes, plasma extravasation of pial venous vessels and leukocyte-endothelial interaction were visualized by intravital microscopy 4 h after LPS administration. Plasma cytokine levels of IL1-β, IL-6, IFN-γ, TNF-α and KC/GRO were evaluated after IVM. A significant plasma extravasation of the pial venous vessels was found in endotoxemia rats compared to control animals. In addition, a significantly increased number of leukocytes adherent to the pial venous endothelium was observed in septic animals. Endotoxemia also induced a significant elevation of plasma cytokine levels of IL1-β, IL-6, IFN-γ, TNF-α and KC/GRO. Endotoxemia increased permeability in the brain pial vessels accompanied by an increase of leukocyte-endothelium interactions and an increase of inflammatory cytokines in the plasma.
- MeSH
- buněčná adheze MeSH
- cévní endotel metabolismus patofyziologie MeSH
- endotoxemie metabolismus patofyziologie MeSH
- interferon gama krev MeSH
- interleukin-1beta krev MeSH
- interleukin-6 krev MeSH
- kapilární permeabilita MeSH
- krysa rodu rattus MeSH
- leukocyty fyziologie MeSH
- mikrocévy metabolismus patofyziologie MeSH
- mikrocirkulace fyziologie MeSH
- mozek krevní zásobení MeSH
- mozkové žíly metabolismus MeSH
- potkani inbrední LEW MeSH
- TNF-alfa krev MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
