Kelly Jane Crossley

Delaying cord clamping until ventilation onset improves cardiovascular function at birth in preterm lambs

•  Delayed cord clamping improves circulatory stability in preterm infants at birth, but the underlying reason is not known. •  In a new preterm lamb study we investigated whether delayed cord clamping until ventilation had been initiated improved pulmonary, cardiovascular and cerebral haemodynamic stability. •  We demonstrated that ventilation prior to cord clamping markedly improves cardiovascular function by increasing pulmonary blood flow before the cord is clamped, thus further stabilising the cerebral haemodynamic transition. •  These results show that delaying cord clamping until after ventilation onset leads to a smoother transition to newborn life, and probably underlies previously demonstrated benefits of delayed cord clamping.

Positive End Expiratory Pressure during Resuscitation of Premature Lambs Rapidly Improves Blood Gases without Adversely Affecting Arterial Pressure

Positive end expiratory pressure (PEEP) is important for neonatal ventilation but is not considered in guidelines for resuscitation. Our aim was to investigate the effects of PEEP on cardiorespiratory parameters during resuscitation of very premature lambs delivered by hysterotomy at ∼125 d gestation (term ∼147 d). Before delivery, they were intubated and lung fluid was drained. Immediately after delivery, they were ventilated with a Dräger Babylog plus ventilator in volume guarantee mode with a tidal volume of 5 mL/kg. Lambs were randomized to receive 0, 4, 8, or 12 cm H2O of PEEP. They were ventilated for a 15-min resuscitation period followed by 2 h of stabilization at the same PEEP. Tidal volume, peak inspiratory pressure, PEEP, arterial pressure, oxygen saturation, and blood gases were measured regularly, and respiratory system compliance and alveolar/ arterial oxygen differences were calculated. Lambs that received 12 cm H2O of PEEP died from pneumothoraces; all others survived without pneumothoraces. Oxygenation was significantly improved by 8 and 12 cm H2O of PEEP compared with 0 and 4 cm H2O of PEEP. Lambs with 0 PEEP did not oxygenate adequately. The compliance of the respiratory system was significantly higher at 4 and 8 cm H2O of PEEP than at 0 PEEP. There were no significant differences in partial pressure of carbon dioxide in arterial blood between groups. Arterial pressure was highest with 8 cm H2O of PEEP, and there was no cardiorespiratory compromise at any level of PEEP. Applying PEEP during resuscitation of very premature infants might be advantageous and merits further investigation.

Early biomarkers and potential mediators of ventilation-induced lung injury in very preterm lambs

BackgroundBronchopulmonary dysplasia (BPD) is closely associated with ventilator-induced lung injury (VILI) in very preterm infants. The greatest risk of VILI may be in the immediate period after birth, when the lungs are surfactant deficient, still partially filled with liquid and not uniformly aerated. However, there have been very few studies that have examined this immediate post-birth period and identified the initial injury-related pathways that are activated. We aimed to determine if the early response genes; connective tissue growth factor (CTGF), cysteine rich-61 (CYR61) and early growth response 1 (EGR1), were rapidly induced by VILI in preterm lambs and whether ventilation with different tidal volumes caused different inflammatory cytokine and early response gene expression.MethodsTo identify early markers of VILI, preterm lambs (132 d gestational age; GA, term ~147 d) were resuscitated with an injurious ventilation strategy (VT 20 mL/kg for 15 min) then gently ventilated (5 mL/kg) for 15, 30, 60 or 120 min (n = 4 in each). To determine if early response genes and inflammatory cytokines were differentially regulated by different ventilation strategies, separate groups of preterm lambs (125 d GA; n = 5 in each) were ventilated from birth with a VT of 5 (VG5) or 10 mL/kg (VG10) for 135 minutes. Lung gene expression levels were compared to levels prior to ventilation in age-matched control fetuses.ResultsCTGF, CYR61 and EGR1 lung mRNA levels were increased ~25, 50 and 120-fold respectively (p < 0.05), within 30 minutes of injurious ventilation. VG5 and VG10 caused significant increases in CTGF, CYR61, EGR1, IL1-β, IL-6 and IL-8 mRNA levels compared to control levels. CTGF, CYR61, IL-6 and IL-8 expression levels were higher in VG10 than VG5 lambs; although only the IL-6 and CYR61 mRNA levels reached significance.ConclusionCTGF, CYR61 and EGR1 may be novel early markers of lung injury and mechanical ventilation from birth using relatively low tidal volumes may be less injurious than using higher tidal volumes.

Dynamic changes in the direction of blood flow through the ductus arteriosus at birth

Major cardiovascular changes occur at birth, including increased pulmonary blood flow (PBF) and closure of the ductus arteriosus (DA), which acts as a low resistance shunt between the fetal pulmonary and systemic circulations. Although the pressure gradient between these circulations reverses after birth, little is known about DA blood flow changes and whether reverse DA flow contributes to PBF after birth. Our aim was to describe the changes in PBF and DA flow before, during and after the onset of pulmonary ventilation at birth. Flow probes were implanted on the left pulmonary artery (LPA) and DA in preterm fetal sheep (n= 8) ∼3 days before they were delivered and ventilated. Blood flow was measured in the LPA and DA, before and after umbilical cord occlusion (UCO) and for 2 h after ventilation onset. Following UCO, DA flow decreased from 534 ± 57 ml min−1 to 237 ± 29 ml min−1 which reflected a similar reduction in right ventricular output. Within 5 min of ventilation onset, PBF increased from 11 ± 6 ml min−1 to 230 ± 13 ml min−1 whereas DA flow decreased to −172 ± 54 ml min−1; negative values indicate reverse DA flow (left‐to‐right shunting). Reverse flow through the DA contributed up to 50% of total PBF at 30 min and a decrease in this contribution accounted for 71 ± 13% of the time‐related decrease in PBF after birth. DA blood flow is very dynamic after birth and depends upon the pressure gradient between the pulmonary and systemic circulations. Following ventilation, reverse DA flow provided a significant contribution to total PBF after birth.

Characterisation of GABAA receptors in fetal, neonatal and adult ovine brain: region and age related changes and the effects of allopregnanolone

Progesterone metabolites acting via GABA(A) receptors suppress central nervous system (CNS) activity. The aim of the present study was to examine binding characteristics of GABA(A) receptors in fetal, newborn and adult sheep brains using [(35)S]TBPS, and to determine the effects of allopregnanolone on this binding. Receptor affinity (K(D)) and density (B(MAX)) in the brainstem were not different in fetal, newborn (1-2 days old) and adult brains. In the hypothalamus K(D) and B(MAX) increased significantly in the fetus between 85 and 128 days gestation, and were then similar to postnatal and adult values. In the frontal cortex K(D) and B(MAX) increased progressively between 85 days and term ( approximately 147 days gestation), and were then not different from postnatal and adult values. The K(i) values for the GABA(A) receptor antagonist picrotoxin was similar at all ages. Allopregnanolone inhibited [(35)S]TBPS binding in the presence of 5 microM GABA, but enhanced binding in the absence of GABA. These results show that (i), functional GABA(A) receptors are present in the fetal brain from at least 85 days gestation; (ii), 3alpha-pregnane steroids modify receptor affinity in the late gestation fetal brain; and (iii) there are region-specific changes in GABA(A) receptor binding parameters. Steroid modulation of the GABA(A) receptor in the fetal brain is likely to influence fetal CNS activity in late gestation.

Intrauterine inflammation causes pulmonary hypertension and cardiovascular sequelae in preterm lambs

Chorioamnionitis increases the risk and severity of persistent pulmonary hypertension of the newborn in preterm infants. Exposure of preterm fetal lambs to intra-amniotic (IA) lipopolysaccharide (LPS) induces chorioamnionitis, causes hypertrophy of pulmonary resistance arterioles, and alters expression of pulmonary vascular growth proteins. We investigated the cardiopulmonary and systemic hemodynamic consequences of IA LPS in preterm lambs. Pregnant ewes received IA injection of LPS (n=6) or saline (controls; n=8) at 122 days gestation, 7 days before exteriorization, instrumentation, and delivery of the fetus with pulmonary and systemic flow probes and catheters at 129 days gestation. Newborn lambs were ventilated, targeting a tidal volume of 6-7 ml/kg and a positive end-expiratory pressure (PEEP) of 4 cmH2O. At 30 min, all lambs underwent a PEEP challenge: PEEP was increased by 2 cmH2O at 10-min intervals to 10 cmH2O and then decreased similarly to 4 cmH2O. Ventilation parameters, arterial blood flows, and pressures were recorded in real-time for 90 min. LPS lambs had higher total protein in bronchoalveolar lavage fluid (P<0.002), increased medial thickness of arteriolar walls (P=0.013), and right ventricular hypertrophy (P=0.012). Compared with controls, LPS lambs had worse oxygenation (P<0.001), decreased pulmonary blood flow (P=0.05), and higher pulsatility index (P<0.001) and pulmonary (P<0.001) and systemic arterial pressures (P=0.005) than controls. Intra-amniotic LPS increased right-to-left shunting across the ductus arteriosus (P=0.018) and decreased left ventricular output (P<0.001). We conclude that inflammation and pulmonary remodeling induced by IA LPS adversely alters pulmonary hemodynamics with subsequent cardiovascular and systemic sequelae, which may predispose the preterm lamb to persistent pulmonary hypertension of the newborn.

Ventilation of the Very Immature Lung In Utero Induces Injury and BPD-Like Changes in Lung Structure in Fetal Sheep

Preterm infants are at high risk of developing ventilator-induced lung injury (VILI), which contributes to bronchopulmonary dysplasia. To investigate causes of VILI, we have developed an animal model of in utero ventilation (IUV). Our aim was to characterize the effects of IUV on the very immature lung, in the absence of nonventilatory factors that could contribute to lung pathology. Fetal sheep were ventilated in utero at 110 d gestation for 1, 6, or 12 h (two groups; n = 5 each). Lung tissue was collected at 12 h after initiating IUV in the 1, 6, and one 12 h IUV groups. Lung liquid was replaced in the second 12 h IUV group and tissues collected at 117 d. Operated, nonventilated 110 and 117 d fetuses were controls. IUV reduced secondary septal crest densities, simplified distal airsacs, caused abnormal collagen and elastin deposition, and stimulated myofibroblast differentiation and cellular proliferation. IUV causes VILI in very immature lungs in the absence of other complicating factors and reproduces bronchopulmonary dysplasia -like changes in lung morphology. IUV offers a novel method for dissociating VILI from other iatrogenic factors that could contribute to altered lung development caused by VILI.

Ventilation onset prior to umbilical cord clamping (physiological-based cord clamping) improves systemic and cerebral oxygenation in preterm lambs.

Background As measurement of arterial oxygen saturation (SpO2) is common in the delivery room, target SpO2 ranges allow clinicians to titrate oxygen therapy for preterm infants in order to achieve saturation levels similar to those seen in normal term infants in the first minutes of life. However, the influence of the onset of ventilation and the timing of cord clamping on systemic and cerebral oxygenation is not known. Aim We investigated whether the initiation of ventilation, prior to, or after umbilical cord clamping, altered systemic and cerebral oxygenation in preterm lambs. Methods Systemic and cerebral blood-flows, pressures and peripheral SpO2 and regional cerebral tissue oxygenation (SctO2) were measured continuously in apnoeic preterm lambs (126±1 day gestation). Positive pressure ventilation was initiated either 1) prior to umbilical cord clamping, or 2) after umbilical cord clamping. Lambs were monitored intensively prior to intervention, and for 10 minutes following umbilical cord clamping. Results Clamping the umbilical cord prior to ventilation resulted in a rapid decrease in SpO2 and SctO2, and an increase in arterial pressure, cerebral blood flow and cerebral oxygen extraction. Ventilation restored oxygenation and haemodynamics by 5–6 minutes. No such disturbances in peripheral or cerebral oxygenation and haemodynamics were observed when ventilation was initiated prior to cord clamping. Conclusion The establishment of ventilation prior to umbilical cord clamping facilitated a smooth transition to systemic and cerebral oxygenation following birth. SpO2 nomograms may need to be re-evaluated to reflect physiological management of preterm infants in the delivery room.

Single Sustained Inflation followed by Ventilation Leads to Rapid Cardiorespiratory Recovery but Causes Cerebral Vascular Leakage in Asphyxiated Near-Term Lambs

Background A sustained inflation (SI) rapidly restores cardiac function in asphyxic, bradycardic newborns but its effects on cerebral haemodynamics and brain injury are unknown. We determined the effect of different SI strategies on carotid blood flow (CaBF) and cerebral vascular integrity in asphyxiated near-term lambs. Methods Lambs were instrumented and delivered at 139 ± 2 d gestation and asphyxia was induced by delaying ventilation onset. Lambs were randomised to receive 5 consecutive 3 s SI (multiple SI; n = 6), a single 30 s SI (single SI; n = 6) or conventional ventilation (no SI; n = 6). Ventilation continued for 30 min in all lambs while CaBF and respiratory function parameters were recorded. Brains were assessed for gross histopathology and vascular leakage. Results CaBF increased more rapidly and to a greater extent during a single SI (p = 0.01), which then decreased below both other groups by 10 min, due to a higher cerebral oxygen delivery (p = 0.01). Blood brain barrier disruption was increased in single SI lambs as indicated by increased numbers of blood vessel profiles with plasma protein extravasation (p = 0.001) in the cerebral cortex. There were no differences in CaBF or cerebral oxygen delivery between the multiple SI and no SI lambs. Conclusions Ventilation with an initial single 30 s SI improves circulatory recovery, but is associated with greater disruption of blood brain barrier function, which may exacerbate brain injury suffered by asphyxiated newborns. This injury may occur as a direct result of the initial SI or to the higher tidal volumes delivered during subsequent ventilation.

Blood gases and pulmonary blood flow during resuscitation of very preterm lambs treated with antenatal betamethasone and/or curosurf : Effect of positive end-expiratory pressure

Resuscitation of very premature lambs with positive end-expiratory pressure (PEEP) improves oxygenation and reduces pulmonary blood flow (PBF). However, the effects of PEEP on blood gases and PBF have not been studied in preterm lambs receiving antenatal corticosteroids or postnatal surfactant. Lambs were delivered at 125 d of gestation (term 147 d) and ventilated with a tidal volume (VT) of 5 mL/kg using different levels of PEEP. Four treatment groups were studied: (1) antenatal betamethasone 24 and 36 h before delivery; (2) postnatal Curosurf; (3) antenatal betamethasone and postnatal Curosurf; (4) untreated controls. Blood gases, PBF, and ventilator parameters were recorded during the first 2 h. Increasing PEEP improved oxygenation even after antenatal betamethasone and postnatal Curosurf, without adverse effects on arterial Pco2. Increasing PEEP reduced PBF; this effect was not altered by betamethasone and/or Curosurf. In very preterm lambs ventilated with fixed VT, increasing levels of PEEP improved oxygenation after antenatal glucocorticoids and/or postnatal surfactant. These treatments do not alter the deleterious effects of high levels of PEEP on PBF.