Dorothy Hehre

Serial determination of pulmonary function in infants with chronic lung disease

Pulmonary function was measured in 39 infants with chronic lung disease who had required mechanical ventilation starting during the first week of life for a median of 9 days (range 1 to 46 days) and supplemental oxygen for a median of 48 days (range 28-162 days). Their mean birth weight was 1140 g (range 550 to 2325 g), and mean gestational age 29.8 weeks (range 26 to 37 weeks). Ventilation was measured by pneumotachography, esophageal pressure through a water-filled feeding tube, and functional residual capacity (FRC) by a modified nitrogen washout technique. Lung compliance, pulmonary conductance, and FRC were determined at 1, 3, 6, 12, 18, 24, and 36 months after birth. Pulmonary function was also determined in 40 normal children, ranging in age from neonates to 5 years, who served as controls. In infants with chronic lung disease, growth in weight and length followed the 10th to 25th percentiles of the normal curve. Minute ventilation and respiratory effort remained elevated throughout the follow-up. FRC per kilogram of body weight was decreased at 1, 3, and 6 months after birth, but thereafter was in the normal range. FRC increased in proportion to weight at the same rate as in the controls. Lung compliance was only half of normal at 1 month, increased with growth in close correlation with weight, and was approximately 80% of normal at the end of follow-up. Pulmonary conductance was 50% of normal at 1 month, increased little during the first 6 months, but reached 85% of normal at 3 years of age. There was no evidence of gas trapping. These results indicate that in infants with chronic lung disease after mechanical ventilation, lung volume increases normally, probably by formation of new alveoli, which also leads to improvement in lung compliance. Airway growth is slow during the first 6 months after birth, but the subsequent faster growth leads to conductance values close to normal at 3 years of age.

Inhibition of the SDF-1/CXCR4 Axis Attenuates Neonatal Hypoxia-Induced Pulmonary Hypertension

Exposure of the neonatal lung to chronic hypoxia produces significant pulmonary vascular remodeling, right ventricular hypertrophy, and decreased lung alveolarization. Given recent data suggesting that stem cells could contribute to pulmonary vascular remodeling and right ventricular hypertrophy, we tested the hypothesis that blockade of SDF-1 (stromal cell–derived factor 1), a key stem cell mobilizer or its receptor, CXCR4 (CXC chemokine receptor 4), would attenuate and reverse hypoxia-induced cardiopulmonary remodeling in newborn mice. Neonatal mice exposed to normoxia or hypoxia were randomly assigned to receive daily intraperitoneal injections of normal saline, AMD3100, or anti–SDF-1 antibody from postnatal day 1 to 7 (preventive strategy) or postnatal day 7 to 14 (therapeutic strategy). As compared to normal saline, inhibition of the SDF-1/CXCR4 axis significantly improved lung alveolarization and decreased pulmonary hypertension, right ventricular hypertrophy, vascular remodeling, vascular cell proliferation, and lung or right ventricular stem cell expressions to near baseline values. We therefore conclude that the SDF-1/CXCR4 axis both prevents and reverses hypoxia-induced cardiopulmonary remodeling in neonatal mice, by decreasing progenitor cell recruitment to the pulmonary vasculature, as well as by decreasing pulmonary vascular cell proliferation. These data offer novel insights into the role of the SDF-1/CXCR4 axis in the pathogenesis of neonatal hypoxia-induced cardiopulmonary remodeling and have important therapeutic implications.

Functional residual capacity in normal neonates and children up to 5 years of age determined by a N2 washout method

ABSTRACT. Functional residual capacity (FRC) was determined in 50 infants by a simplified N2 washout method. Fourteen infants were preterm, four full-term newborns and the rest were 1 month to 5 yr of age. Weight ranged from 1.19 to 25.8 kg. The method gave well reproducible values with a mean coefficient of variation of 3.9%. The FRC values are equally well correlated to weight and length (r = 0.98). The correlation with weight is linear, intercepting the x axis (FRC = 0) at a weight of 480 g, the one with length is best described by a power curve. The course of the regression lines reflects the observation that FRC per kg weight or per cm length is lower in neonates than in larger infants. The FRC measurements are in the same range as values obtained by other investigators using the N2 washout or He-dilution techniques. The values are significantly smaller than thoracic gas volume measurements obtained by plethysmography. This difference may be due to air trapping or to possible methodological problems with the plethysmographic technique. The data demonstrate that FRC can be measured easily and accurately in preterm and older infants using a N2 washout technique.

A simple method for measuring functional residual capacity by N2 washout in small animals and newborn infants

ABSTRACT: An open circuit N2 washout technique is described for the determination of functional residual capacity in infants. Either 100% O2 or any oxygen/helium mixture can be used as the washing gas. The subject breathes the washing gas through a T-tube and the washed out nitrogen is mixed with this gas in a mixing chamber, placed into the exhalation part of the circuit. The N2 concentration of the mixed gas is analyzed continuously, and the concentration signal is electronically integrated over time. Calibration of the system is accomplished by injecting known amounts of nitrogen or room air into the circuit. The gas flow through the system must remain constant and is adjusted to approximate peak inspiratory flow of the infant. In vitro testing of the system showed that the technique gives reproducible values (coefficient of variance <1.0%) and that the integrated signal output has a close linear correlation with the amount of N2 washed out (r = 0.99). In vivo measurements in 10 cats confirmed the accuracy and reproducibility of the method when compared with N2 collection. The technical advantages of the system are simplicity of components, absence of valves, easy calibration, low dead space, and no need to collect or measure expired gases. For the infant this means no added resistance during washout and no risk of hypoxia, hyperoxia, or hypercapnea. In the presence of pulmonary disease and poor gas mixing the washout period can be prolonged as needed. There is no lower limit of weight or size for functional residual capacity measurements in small infants or animals.

Long-term reparative effects of mesenchymal stem cell therapy following neonatal hyperoxia-induced lung injury.

Background:Mesenchymal stem cell (MSC) therapy may prevent neonatal hyperoxia-induced lung injury (HILI). There are, however, no clear data on the therapeutic efficacy of MSC therapy in established HILI, the duration of the reparative effects, and the exact mechanisms of repair. The main objective of this study was to evaluate whether the long-term reparative effects of a single intratracheal (IT) dose of MSCs or MSC-conditioned medium (CM) are comparable in established HILI.Methods:Newborn rats exposed to normoxia or hyperoxia from postnatal day (P)2)–P16 were randomized to receive IT MSCs, IT CM, or IT placebo (PL) on P9. Alveolarization and angiogenesis were evaluated at P16, P30, and P100.Results:At all time periods, there were marked improvements in alveolar and vascular development in hyperoxic pups treated with MSCs or CM as compared with PL. This was associated with decreased expression of inflammatory mediators and an upregulation of angiogenic factors. Of note, at P100, the improvements were more substantial with MSCs as compared with CM.Conclusion:These data suggest that acute effects of MSC therapy in HILI are mainly paracrine mediated; however, optimum long-term improvement following HILI requires treatment with the MSCs themselves or potentially repetitive administration of CM.

Gas trapping with high-frequency ventilation: Jet versus oscillatory ventilation

Gas trapping was evaluated during high-frequency jet ventilation (HFJV) and high-frequency oscillatory ventilation (HFOV) in nine adult rabbits under basal conditions and after instillation of a mixture of 20% human meconium (2 mL/kg). The anesthetized animals underwent tracheostomy and were placed inside a body plethysmograph. Respiratory compliance and resistance were calculated from airway pressure and simultaneous flow, and volume was measured with a pneumotachograph. Gas trapping was measured as the change in volume observed in the plethysmograph after clamping the jet or the oscillatory line at respiratory rates of 10 and 15 Hz and tidal volumes of 1.0 and 2.0 mL/kg. Mean airway pressure was similar with both ventilators. Inspiratory/expiratory ratios were 1:4 at 10 Hz and 1:2 at 15 Hz with HFJV, and 1:1 during HFOV. Under all conditions, gas trapping was significantly greater with HFJV than with HFOV. More gas trapping was observed with higher tidal volume (2 mL/kg) and respiratory rate (15 Hz) during HFJV, before and after meconium instillation. After meconium instillation, gas trapping during HFJV at 15 Hz and tidal volume 2 mL/kg decreased significantly (32.7 +/- 10.4 to 24.9 +/- 10.3; P less than 0.05), compared with basal conditions. This finding may be explained by the shorter time constant of the respiratory system after meconium instillation (0.118 vs 0.083 seconds, P less than 0.01). Thus gas trapping was significantly greater with HFJV than with HFOV, a difference most likely related to the active expiratory phase of HFOV.

Connective Tissue Growth Factor Antibody Therapy Attenuates Hyperoxia-Induced Lung Injury in Neonatal Rats

Despite recent advances in neonatal intensive care and surfactant therapy, bronchopulmonary dysplasia (BPD) continues to be one of the most common long-term pulmonary complications associated with preterm birth. Clinical efforts to prevent and treat BPD have been largely unsuccessful due to its multifactorial nature and poorly understood disease process. Connective tissue growth factor (CTGF) is a matricellular protein that plays an important role in tissue development and remodeling. Previous studies have demonstrated that hyperoxia exposure up-regulates CTGF expression in neonatal rat lungs. Whether CTGF overexpression plays a role in the pathogenesis of BPD, and whether CTGF antagonism has a therapeutic potential for BPD, are unknown. In the present study, we examined CTGF expression in lung autopsy specimens from patients with BPD and control subjects with no BPD. We assessed the effect of a CTGF-neutralizing monoclonal antibody (CTGF Ab) on preventing hyperoxia-induced lung injury in neonatal rats. Our study demonstrates that CTGF expression is increased in BPD lungs. In newborn rats, exposure to 90% oxygen for 14 days resulted in activation of β-catenin signaling, decreased alveolarization and vascular development, and physiological and histological evidence of pulmonary hypertension (PH). However, treatment with CTGF Ab prevented β-catenin signaling activation, improved alveolarization and vascular development, and attenuated PH during hyperoxia. These data indicate that CTGF-β-catenin signaling plays a critical role in the pathogenesis of experimental BPD. CTGF antagonism may offer a novel therapeutic strategy to alleviate BPD and PH in neonates.

CTGF disrupts alveolarization and induces pulmonary hypertension in neonatal mice: implication in the pathogenesis of severe bronchopulmonary dysplasia

The pathological hallmarks of bronchopulmonary dysplasia (BPD), one of the most common long-term pulmonary complications associated with preterm birth, include arrested alveolarization, abnormal vascular growth, and variable interstitial fibrosis. Severe BPD is often complicated by pulmonary hypertension characterized by excessive pulmonary vascular remodeling and right ventricular hypertrophy that significantly contributes to the mortality and morbidity of these infants. Connective tissue growth factor (CTGF) is a multifunctional protein that coordinates complex biological processes during tissue development and remodeling. We have previously shown that conditional overexpression of CTGF in airway epithelium under the control of the Clara cell secretory protein promoter results in BPD-like architecture in neonatal mice. In this study, we have generated a doxycycline-inducible double transgenic mouse model with overexpression of CTGF in alveolar type II epithelial (AT II) cells under the control of the surfactant protein C promoter. Overexpression of CTGF in neonatal mice caused dramatic macrophage and neutrophil infiltration in alveolar air spaces and perivascular regions. Overexpression of CTGF also significantly decreased alveolarization and vascular development. Furthermore, overexpression of CTGF induced pulmonary vascular remodeling and pulmonary hypertension. Most importantly, we have also demonstrated that these pathological changes are associated with activation of integrin-linked kinase (ILK)/glucose synthesis kinase-3β (GSK-3β)/β-catenin signaling. These data indicate that overexpression of CTGF in AT II cells results in lung pathology similar to those observed in infants with severe BPD and that ILK/GSK-3β/β-catenin signaling may play an important role in the pathogenesis of severe BPD.

Targeting glycogen synthase kinase-3β to prevent hyperoxia-induced lung injury in neonatal rats.

The pathological hallmarks of bronchopulmonary dysplasia (BPD), a chronic lung disease of premature infants, include inflammation, arrested alveolarization, and dysregulated angiogenesis. Severe BPD is often complicated by pulmonary hypertension (PH) that significantly increases morbidity and mortality. Glycogen synthase kinase (GSK)-3β plays a pivotal role in embryonic development, cell proliferation and survival, and inflammation by modulating multiple signaling pathways, particularly the nuclear transcription factor, NF-κB, and Wnt/β-catenin pathways. Aberrant GSK-3β signaling is linked to BPD. We tested the hypothesis that inhibition of GSK-3β is beneficial in preventing hyperoxia-induced neonatal lung injury, an experimental model of BPD. Newborn rats were exposed to normoxia or hyperoxia (90% oxygen), and received daily intraperitoneal injections of placebo (DMSO) or SB216763, a specific pharmacological inhibitor of GSK-3β, for 14 days. Hyperoxia exposure in the presence of the placebo increased GSK-3β phosphorylation, which was correlated with increased inflammation, decreased alveolarization and angiogenesis, and increased pulmonary vascular remodeling and PH. However, treatment with SB216763 decreased phosphorylation of NF-κB p65, expression of monocyte chemotactic protein-1, and lung inflammation during hyperoxia. Furthermore, treatment with the GSK-3β inhibitor also improved alveolarization and angiogenesis, and decreased pulmonary vascular remodeling and PH. These data indicate that GSK-3β signaling plays an important role in the pathogenesis of hyperoxia-induced neonatal lung injury, and that inhibition of GSK-3β is beneficial in preventing inflammation and protecting alveolar and vascular structures during hyperoxia. Thus, targeting GSK-3β signaling may offer a novel strategy to prevent and treat preterm infants with BPD.

Brainstem Amino Acid Neurotransmitters and Ventilatory Response to Hypoxia in Piglets

The ventilatory response to hypoxia is influenced by the balance between inhibitory (GABA, glycine, and taurine) and excitatory (glutamate and aspartate) brainstem amino acid (AA) neurotransmitters. To assess the effects of AA in the nucleus tractus solitarius (NTS) on the ventilatory response to hypoxia at 1 and 2 wk of age, inhibitory and excitatory AA were sampled by microdialysis in unanesthetized and chronically instrumented piglets. Microdialysis samples from the NTS area were collected at 5-min intervals and minute ventilation (VE), arterial blood pressure (ABP), and arterial blood gases (ABG) were measured while the animals were in quiet sleep. A biphasic ventilatory response to hypoxia was observed in wk 1 and 2, but the decrease in VE at 10 and 15 min was more marked in wk 1. This was associated with an increase in inhibitory AA during hypoxia in wk 1. Excitatory AA levels were elevated during hypoxia in wk 1 and 2. Changes in ABP, pH, and ABG during hypoxia were not different between weeks. These data suggest that the larger depression in the ventilatory response to hypoxia observed in younger piglets is mediated by predominance of the inhibitory AA neurotransmitters, GABA, glycine, and taurine, in the NTS.