Search

Objective: To describe the development of an artificial placenta (AP) system in sheep with learning curve and main bottlenecks to allow survival up to one week. Methods: A total of 28 fetal sheep were transferred to an AP system at 110–115 days of gestation. The survival goal in the AP system was increased progressively in three consecutive study groups: 1–3 h (n = 8), 4–24 h (n = 10) and 48–168 h (n = 10). Duration of cannulation procedure, technical complications, pH, lactate, extracorporeal circulation (EC) circuit flows, fetal heart rate, and outcomes across experiments were compared. Results: There was a progressive reduction in cannulation complications (75%, 50% and 0%, p = 0.004), improvement in initial pH (7.20 ± 0.06, 7.31 ± 0.04 and 7.33 ± 0.02, p = 0.161), and increment in the rate of experiments reaching survival goal (25%, 70% and 80%, p = 0.045). In the first two groups, cannulation accidents, air bubbles in the extracorporeal circuit, and thrombotic complications were the most common cause of AP system failure. Conclusions: Achieving a reproducible experimental setting for an AP system is extremely challenging, time- and effort-consuming, and requires a highly multidisciplinary team. As a result of the learning curve, we achieved reproducible transition and survival up to 7 days. Extended survival requires improving instrumentation with custom-designed devices. [ABSTRACT FROM AUTHOR]

Objective: To describe the development of an artificial placenta (AP) system in sheep with learning curve and main bottlenecks to allow survival up to one week. Methods: A total of 28 fetal sheep were transferred to an AP system at 110–115 days of gestation. The survival goal in the AP system was increased progressively in three consecutive study groups: 1–3 h (n = 8), 4–24 h (n = 10) and 48–168 h (n = 10). Duration of cannulation procedure, technical complications, pH, lactate, extracorporeal circulation (EC) circuit flows, fetal heart rate, and outcomes across experiments were compared. Results: There was a progressive reduction in cannulation complications (75%, 50% and 0%, p = 0.004), improvement in initial pH (7.20 ± 0.06, 7.31 ± 0.04 and 7.33 ± 0.02, p = 0.161), and increment in the rate of experiments reaching survival goal (25%, 70% and 80%, p = 0.045). In the first two groups, cannulation accidents, air bubbles in the extracorporeal circuit, and thrombotic complications were the most common cause of AP system failure. Conclusions: Achieving a reproducible experimental setting for an AP system is extremely challenging, time- and effort-consuming, and requires a highly multidisciplinary team. As a result of the learning curve, we achieved reproducible transition and survival up to 7 days. Extended survival requires improving instrumentation with custom-designed devices.

Background: We created abdominal wall defects (AWD) in fetal lambs to investigate possible causes of scoliosis. Methods: We incised the upper abdominal wall (including Rectus) in 60-day gestation fetal lambs, from the midline to either the right (Group A) or left (Group B) costal margin, in 14 lambs carried by 7 ewes. They were delivered by cesarean section at term (about 145 days). Scoliosis was evaluated by anterio-posterior X-rays, determining the Cobb angle. Results: Four fetuses in Group A and 3 in Group B survived. There were 3 successful AWD lambs Group A and 2 in Group B. One lamb in each group survived with the AWD covered with a thick capsule. The convexity of spinal curve was the direction of scoliosis. Right scoliosis was only seen in the 4 Group A lambs. Left scoliosis was only seen in Group B lambs (2/3, 67%). The mean Cobb angle was 41.7 ± 11.5° in Group A and in Group B the Cobb angles were 59.6o and 60.6°. Overall, 4/5 lambs with organ prolapse (80%) and both lambs without organ prolapse had scoliosis. Conclusion: Muscle imbalance may contribute to the development of scoliosis in a fetal lamb AWD model. [ABSTRACT FROM AUTHOR]

Background and Aims: The Extra-Uterine Environment for Neonatal Development (EXTEND) aims to avoid the complications of prematurity, such as NEC. Our goal was to determine if bowel development occurs normally in EXTEND-supported lambs, with specific emphasis on markers of immaturity associated with NEC. Methods: We compared terminal ileum from 17 pre-term lambs supported on EXTEND for 2- 4 weeks to bowel from age-matched fetal lambs that developed in utero. We evaluated morphology, markers of epithelial integrity and maturation, enteric nervous system structure, and bowel motility. Results: EXTEND-supported lamb ileum had normal villus height, crypt depth, density of mucin-containing goblet cells, and enteric neuron density. Expression patterns for I-FABP, activated caspase-3 and EGFR were normal in bowel epithelium. Transmural resistance assessed in Ussing chambers was normal. Bowel motility was also normal as assessed by ex vivo organ bath and video imaging. However, Peyer’s patch organization did not occur normally in EXTEND ileum, resulting in fewer circulating B cells in experimental animals. Conclusion: EXTEND supports normal ileal epithelial and enteric nervous system maturation in pre-term lambs. The classic morphologic changes and cellular expression profiles associated with NEC are not seen. However, immune development within the EXTEND supported lamb bowel does not progress normally.

In recent decades, new research into the developmental defects and pathophysiological basis of congenital diaphragmatic hernia has revealed opportunities for the development of innovative therapies. Importantly, the use of animal models to represent this anomaly in the laboratory has resulted in the discovery of many important genetic, epigenetic and other molecular contributors to this condition. In this review, the most commonly used and newly devised animal models of CDH are presented to familiarize the reader with the latest innovations in the basic sciences.

Despite advances in neonatology, the hypoxic–ischemic injury in the perinatal period remains the single most important cause of brain injury in the newborn, leading to death or lifelong sequelae. Using a sheep model of intrauterine asphyxia, we evaluated the correlation between reactive oxygen species (ROS) overproduction, cytokine expression, and apoptotic cell death. Fetal lambs were assigned to sham group, nonasphyctic animals; and hypoxia–ischemia (HI) group, lambs subjected to 60 minutes of HI) by partial cord occlusion and sacrificed 3 hours later. Different brain regions were separated to quantify the number of apoptotic cells and the same territories were dissociated for flow cytometry studies. Our results suggest that the overproduction of ROS and the early increase in cytokine production after HI in fetal lambs correlate in a significant manner with the apoptotic index, as well as with each brain region evaluated. [ABSTRACT FROM AUTHOR]

Abstract: The objective was to investigate the relationship between fetal aortic distension waveforms and fetal hypoxemia and/or acidosis. Aortic distension waveforms were recorded using an echo-tracking system in nine late-gestation catheterized fetal lambs. Under hypoxic conditions induced by inhalation of gas mixture, fetal blood pressure and aortic distension waveforms were recorded. Four parameters, namely peak systolic and end diastolic diameter, amplitude (ΔD) and ratio of ΔD to end diastolic diameter (%ΔD), were obtained, and correlations between these parameters and partial arterial oxygen tension (PaO2) and pH of fetal blood gas were analyzed. These four parameters were compared between the control, hypoxemic and asphyxic groups. The ΔD and %ΔD were significantly correlated with PaO2 and pH using linear regression analysis. Both the ΔD and %ΔD decreased significantly, in a stepwise fashion, in the hypoxemic and asphyxiated groups compared with controls using repeated measured analysis of variance. It was concluded that fetal aortic distension waveforms proved to be a useful tool to detect the deterioration in the fetal circulation secondary to intrauterine hypoxemia/asphyxia. (E-mail: satohs@oitakenbyo.jp) [Copyright &y& Elsevier]

Aim: To investigate which part of the fetal aortic distension waveform is mainly influenced by changes in fetal cardiac contractility and aortic blood pressure. Methods: In acute preparation, aortic distension waveforms were recorded using an echo-tracking system, and aortic and left ventricular pressure waveforms were obtained from six late-gestation catheterized fetal lambs. Dobutamine and angiotensin II were separately infused and the correlations between the maximum value in the first derivative of left ventricle pressure waveforms (Max dP/dt) and fetal blood pressure, and the parameters obtained from aortic distension waveforms were analyzed using linear regression analysis. Results: With a change in cardiac contractility, the maximum value of first derivative of the systolic rising slope in the aortic distension waveform had a positive correlation with Max dP/dt ( r = 0.93, P < 0.0001). With changes in fetal blood pressure, both the amplitude and the ratio of increase from the end diastolic diameter obtained from the aortic distension waveform had a significant positive correlation with aortic blood pressure amplitude ( r = 0.60, P < 0.01; r = 0.61, P < 0.01, respectively). Conclusion: The maximum first derivative of the systolic rising slope in the aortic distension waveform and the amplitude in the aortic distension waveform enable us to non-invasively substitute for fetal cardiac contractility and aortic blood pressure amplitude, respectively. [ABSTRACT FROM AUTHOR]

Objectives: The objectives of this study are firstly to clarify how the Doppler isovolumetric contraction time (ICT) is influenced by arterial oxygen pressure (PaO2) or pH, and secondly to confirm the relationship between the Doppler ICT and myocardial contractility during hypoxemia and/or acidemia in the fetal lamb. Material and Methods: In 12 pregnant ewes, fetal hypoxemia and acidemia were induced by giving ewes a variable mixture of gases for 120 min. The chronological change of fetal PaO2, pH, Doppler ICT and maximum first derivative of the left ventricular pressure waveform (Max dp/dt) was recorded every 30 min. Results: Doppler ICT and PaO2 had no significant regression. On the other hand, Doppler ICT and pH demonstrated a significant negative regression. Moreover, one critical given pH point was indicated with statistical significance at 7.20 and the prolongation of the Doppler ICT was found more markedly in the range below the pH of 7.20 compared with the range above 7.20. A significant negative linear regression was found between the Doppler ICT and the Max dp/dt. Conclusion: The measurement of Doppler ICT enables us to predict severe acidosis and a decrease of myocardial contractility in the fetus. [ABSTRACT FROM AUTHOR]

We measured the velocity and attenuation of audible sound in the isolated lung of the near-term fetal sheep to test the hypothesis that the acoustic properties of the lung provide a measure of the volume of gas it contains. We introduced pseudorandom noise (bandwidth 70 Hz-7 kHz) to one side of the lung and recorded the noise transmitted to the surface immediately opposite, starting with the lung containing only fetal lung liquid and making measurements after stepwise inflation with air until a leak developed. The velocity of sound in the lung fell rapidly from 187 ± 28.2 to 87 ± 3.7 m/s as lung density fell from 0.93 ± 0.01 to 0.75 ± 0.01 g/ml (lung density = lung weight/gas volume plus lung tissue volume). For technical reasons, no estimate of velocity could be made before the first air injection. Thereafter, as lung density fell to 0.35 ± 0.01 g/ml, there was a further decline in velocity to 69.6 ± 4.6 m/s. High-frequency sound was attenuated as lung density decreased from 1.0 to 0.5 g/ml, with little change thereafter down to a density of 0.35 ± 0.01 g/ml. We conclude that both the velocity of audible sound through the lung and the degree to which high-frequency sound is attenuated in the lung provide information on the degree of inflation of the isolated fetal lung, particularly at high lung densities. If studies of sound transmission through the lung in the intact organism were to confirm these findings, the acoustic properties of the lung could provide a means for monitoring lung aeration during mechanical ventilation of newborn infants. [ABSTRACT FROM AUTHOR]

Background: The aim of the current study was to investigate the changes in the power spectral pattern of the heart rate variability of fetal lambs during acute hypoxia and the possible value of power spectral analysis as a quantitative fetal monitoring method.Methods: Acutely instrumented eight fetal lambs in the third trimester of gestation were subjected to reproducible hypoxia by reducing the maternal placental blood flow with complete obstruction of the maternal abdominal aorta for 60 s. Fetal electrocardiographic data 5 min prior to occlusion, 1 min during occlusion and 1, 5, and 10 min after the removal of occlusion were analyzed using power spectral analysis. Differences among the procedural steps were determined by the Friedman test with multiple comparisons using Duncan's multiple-range test. Wilcoxon's rank sum test was used for the comparison between low-and high-frequency power values at each step. p< 0.05 was considered significant.Results: Low-, high-, total-frequency power and low-to-high frequency ratio all significantly increased with hypoxia compared with the baseline state (p < 0.05). High-frequency power remained higher than low-frequency power during the resting state (baseline state, 5 and 10 min after hypoxia).Conclusions: Increased low-frequency power and low-to-high frequency ratio during hypoxia reflects increased sympathetic activity compared with the baseline state. Higher high-frequency power during the resting state compared with low-frequency power reflects active respiratory movement of the fetal lambs near term and increased parasympathetic activity. It appeared possible that power spectral analysis could serve as a useful quantitative tool to monitor the autonomic changes in fetal lambs during hypoxia. [ABSTRACT FROM AUTHOR]

Maternal transplacental administration of sildenafil is being considered for a variety of fetal conditions. Clinical translation also requires evaluation of fetal safety in a higher species, such as the fetal lamb. Experiments with the pregnant ewe are curtailed by minimal transplacental transfer as well as limited access to the fetus. The EXTra-uterine Environment for Neonatal Development (EXTEND) model renders the isolated fetal lamb readily accessible and allows for direct fetal administration of sildenafil. Five fetal lambs were placed on extracorporeal support in the EXTEND device and received continuous intravenous (IV) sildenafil (0.3–0.5–0.7 mg/kg/24hr) for a duration of one to seven days. Plasma sildenafil concentrations were sampled at regular intervals to establish the pharmacokinetic profile using population pharmacokinetic modeling. Serial Doppler ultrasound examination, continuous non-invasive hemodynamic monitoring and blood gas analysis were done to evaluate the pharmacodynamic effects and fetal response. The target concentration range (47–500 ng/mL) was attained with all doses. Sildenafil induced an immediate and temporary reduction of pulmonary vascular resistance, mean arterial pressure and circuit flow, without change in fetal lactate levels and acid-base status. The duration of the systemic effects increased with the dose. Immediate temporary pulmonary vascular and systemic hemodynamic changes induced by sildenafil were biochemically well tolerated by fetal lambs on extracorporeal support, with the 0.5 mg/kg/24 h dose balancing rapid attainment of target concentrations with short-lived systemic effects. None. A literature review was conducted searching online databases (Medline, Embase and Cochrane), using search terms: fetal OR prenatal OR antenatal AND sildenafil, without time-limit and excluding human studies. Where relevant, investigators were contacted in order to avoid duplication of work. Prenatal therapy with sildenafil, a phosphodiesterase-5 inhibitor with vasodilatory and anti-remodeling effects on vascular smooth muscle cells, has been considered for a variety of fetal conditions. One multicenter clinical trial investigating the benefit of sildenafil in severe intrauterine growth restriction (the STRIDER-trial) was halted early due to excess mortality in the sildenafil-exposed arm at one treatment site. Such findings demonstrate the importance of extensive preclinical safety assessment in relevant animal models. Transplacentally administered sildenafil leads to decreased pulmonary arterial muscularization, preventing or reducing the occurrence of pulmonary hypertension in rat and rabbit fetuses with diaphragmatic hernia (DH). Validation of these results in a higher and relevant animal model, e.g. fetal lambs, is the next step to advance clinical translation. We recently demonstrated that, in contrast to humans, transplacental transfer of sildenafil in sheep is minimal, precluding the in vivo study of fetal effects at target concentrations using the conventional pregnant ewe model. We therefore used the extracorporeal support model for fetal lambs, referred to as the EXTra-uterine Environment for Neonatal Development (EXTEND) system, bypassing placental and maternal metabolism, to investigate at what dose the target concentrations are reached, and what the fetal hemodynamic impact and response are. Fetal hemodynamic and metabolic tolerance to sildenafil are a crucial missing element on the road to clinical translation. This is therefore the first study investigating the pharmacokinetics, hemodynamic and biochemical effects of clinical-range concentrations of sildenafil in fetal lambs, free from placental and maternal interference. We demonstrated self-limiting pulmonary vasodilation, a decrease of both systemic arterial pressures and circuit flows, induced by clinical range concentrations of sildenafil, without the development of fetal acidosis. This paves the way for further investigation of prenatal sildenafil in fetal lambs on extracorporeal support. A dose of 0.5 mg/kg/24 h offered the best trade-off between rapid achievement of target concentrations and shortest duration of systemic effects. This is also the first study using the EXTEND as a model for pharmacotherapy during pregnancy. [Display omitted] • Sildenafil was investigated in isolated fetal lambs on extracorporeal support. • Relevant concentrations of sildenafil could be measured. • Sildenafil caused an immediate and temporary pulmonary vasodilation. • Sildenafil was biochemically well-tolerated by the fetal lambs. [ABSTRACT FROM AUTHOR]