Three dimensional structures of pulmonary elastin; airway vs vascular elastin

Elastin is known to occur in the lung parenchyma and pleura as well as in the pulmonary vessels, but no detailed studies of this elastin's linkage between them have been done in three dimensions. For many years we have known that there is abundant elastin in the mammalian lungs, which may be associated with etiology of causing emphysema. We have developed selective casting methods to allow us to determine the location where elastin is found morphologically. The method involves casting either the vasculature via the right ventricle, or the airways via the trachea in the air sacs. Studies of the vasculature were done with the lung inflated to 80% of the vital capacity. The casted lungs were then put in 0.1 N NaOH at 75 degrees C for 48 hours, turning them frequently. THis method removed all non-elastin tissues. The scanning electron microscopy (SEM) was used to reveal the three dimensional pictures of elastin structures from both lung parenchyma and pulmonary vessels. Elastin was seen as fenestrated sheets and some fibers in both the vessels and the airways. Elastin in the two different locations was often interconnected. Studies on 6 dogs, 8 rabbits, and 2 pigs showed no significant species difference at the level of resolution of the SEM, which was used to study the specimens after they had been freeze-dried.

Morphological evidence of pulmonary vascular leakage through gaps observed with casting methods and S.E.M

We have used selective casting methods to separate pulmonary elastin from vascular elastin in the lungs of rabbits, dogs and pigs. The lungs are digested with 0.1 N NaOH at 75 degrees C for 24 approximately 48 hours with frequent turning as the lungs are filled with air to about 80% of the vital capacity prior to the casting which is done at pressure of 20 approximately 50 mmHg. After vascular injections, we saw many small globular bits of casting material well separated from cast vessels and lying in the pulmonary elastin. Surface forces should make the casting material creep along the vessels even if they are not completely filled, so that the spherical shape is the one expected if the case is extruded into the parenchymal space and the air space. We conclude that this suggests that the pulmonary circulation is partially and temporarily 'open' as seen in the spleen and some other organs, rather than a completely 'closed' one as is generally accepted. At least some of these extravasations may be associated with lymphatics, although we have not proved this.