Developmentally regulated changes in extracellular matrix in endothelial and smooth muscle cells in the ductus arteriosus may be related to intimal proliferation.

In the late gestation fetal lamb ductus arteriosus (DA), intimal proliferation is observed, characterized by smooth muscle migration and proliferation in the subendothelium. The nature of changes in the endothelial and smooth muscle extracellular matrix associated with the development of this feature are not known. We assessed the production of glycoproteins (fibronectin, laminin, and type IV collagen) and glycosaminoglycans (GAGs) (hyaluronic acid, heparan sulfate, and chondroitin sulfate) in endothelial and smooth muscle cells harvested from the DA, aorta (Ao), and pulmonary artery of fetal lambs at 100 days gestation, before the appearance of DA intimal proliferation, and at 138 days, when well-developed intimal 'cushions' are seen. In passage 3 cells, glycoprotein synthesis was measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after 48 hours incubation with [35S]methionine, and GAGs were assessed by labeling with [3H] glucosamine and separation on DEAE ion-exchange high performance liquid chromatography. Analyses were carried out on culture medium, cell layer, and solubilized matrix. Fibronectin secretion by DA smooth muscle cells from 100-day lambs was found to be twice that of Ao or pulmonary artery cells. No significant differences were seen in smooth muscle cells from 138-day lambs or when comparing endothelial cells from each of the vascular sites at both gestational ages. As well, there were no DA-specific differences in laminin or type IV collagen. No significant differences in endothelial GAG secretion were observed comparing each vascular site at both gestational ages. Analysis of endothelial-derived matrices, however, revealed increased incorporation of hyaluronic acid in the DA from 100-day lambs, 10-fold that of the pulmonary artery and Ao, and increased heparan sulfate. These differences were still present in cell matrices from late gestation animals, but were less marked. No differences in GAGs were seen when comparing smooth muscle cells. Incubation of 100-day DA and Ao smooth muscle cells with endothelial conditioned medium however, resulted in a 2-fold increase in chondroitin sulfate in DA, compared with Ao. These results indicate that distinct, developmentally regulated patterns of extracellular matrix production are related to vascular site and specific features appear to precede intimal proliferation in the DA.