title: Effect of antenatal tracheal occlusion on lung development in the sheep model of congenital diaphragmatic hernia: a morphometric analysis of pulmonary structure and maturity.
abstract: The incidence of congenital diaphragmatic hernia (CDH) is 1:1,207-5,000, and the condition is associated with high mortality and morbidity, attributed principally to associated pulmonary hypoplasia. Repairing the diaphragmatic defect by antenatal surgery has high mortality, mainly due to premature labor. Antenatal tracheal occlusion, which is achievable by less invasive methods, stimulates lung growth (weight and DNA). However, its effectiveness in reversing structural and maturational abnormalities and its optimal timing requires further investigation. We hypothesized that (1) antenatal tracheal occlusion performed in the lamb model of congenital diaphragmatic hernia will stimulate lung growth and structural development and restore lung structure and maturity toward normal levels by term gestation; (2) effects will be detectable by morphometric measurements of the following parameters: lung volume, ratio of parenchyma to nonparenchyma, volume density of connective tissue within nonparenchyma, ratio of gas exchange tissue to airspace in parenchyma, gas exchange surface area, capillary loading, alveolar/airspace density and alveolar perimeter; (3) effects will be seen in all lobes of the lung; and (4) a greater effect will be observed when tracheal occlusion is performed early rather than late in gestation. Fourteen lambs underwent CDH creation at gestation day 72-74 followed by tracheal occlusion at day 101 (n = 7) or 129 (n = 7). They were delivered by Cesarean section at 143 days (term = 145-149). Lungs were obtained at autopsy, inflation fixed, divided into lobes, and sampled; morphometric analysis was performed. Comparisons were made with previously reported results from control lungs of normal lambs and lambs with untreated CDH. In comparison with untreated lungs, antenatal tracheal occlusion at both times resulted in increased volumes for total lung and lobes, increased volume density of parenchyma and of airspace within parenchyma, and increased gas exchange surface areas. Normal values for gas exchange surface area density, and alveolar density and perimeter were attained and the lungs appeared more mature than non-occluded lungs. Tracheal occlusion earlier in gestation produced a greater effect, achieving greater than normal values for lung volumes and volume densities, whereas the capillary loading value was similar to normal lung. Later occlusion achieved less than normal values for lung volumes and volume densities, with a reduced capillary loading value. We conclude that antenatal tracheal occlusion is capable of reversing structural total lung and lobar hypoplasia and immaturity caused by CDH as determined by morphometrically determined parameters. The effect is greater when tracheal occlusion is performed early rather than late in gestation. The results are encouraging for development of treatment methods for humans with antenatally diagnosed CDH.
authors.affiliation: Department of Histopathology, Child Health Research Institute, Women's and Children's Hospital, Adelaide, North Adelaide, South Australia, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Lipsett
authors.forename: J C
authors.initials: JC
authors.lastname: Cool
authors.forename: S I
authors.initials: SI
authors.lastname: Runciman
authors.forename: W D
authors.initials: WD
authors.lastname: Ford
authors.forename: J D
authors.initials: JD
authors.lastname: Kennedy
authors.forename: A J
authors.initials: AJ
authors.lastname: Martin
mesh_terms: D000818:Animals; D002452:Cell Count; D005069:Evaluation Studies as Topic; D005260:Female; D005325:Fetal Organ Maturity; D006548:Hernia, Diaphragmatic / Q000196:embryology*; D065630:Hernias, Diaphragmatic, Congenital; D008168:Lung / Q000166:cytology / Q000196:embryology* / Q000254:growth & development; D009024:Morphogenesis; D011247:Pregnancy; D011659:Pulmonary Gas Exchange; D011897:Random Allocation; D012756:Sheep; D014132:Trachea / Q000601:surgery*