Schneider A, Schwalb H, Vlodavsky I, Uretzky G
The Joseph Lunenfeld Cardiac Surgery Research Center, Hadassah University Hospital, Jerusalem, Israel.
Clin Mater. 1993;13(1-4):51-5. doi: 10.1016/0267-6605(93)90090-t.
The feature that clearly distinguishes natural blood vessels from their artificial counterparts is the presence of the endothelial cell lining that, besides being non-thrombogenic, is capable of repair and renewal. This study describes a method of coating vascular grafts with a uniform naturally produced subendothelial extracellular matrix (ECM) prior to implantation. Bovine aortic endothelial cells were seeded at a low density on untreated GORE-TEX, GORE-TEX that was precoated with ECM, or GORE-TEX that was first coated with fibronectin and then with ECM. The cells were maintained in the absence of fibroblast growth factor (FGF) and the cell number determined after 7 days in culture. Poor growth was observed on untreated GORE-TEX (6030 +/- 1030 cells/well), a 10-fold improved growth was observed on ECM coated GORE-TEX (59900 +/- 1560 cells/well) and best results were obtained on fibronectin-ECM precoated GORE-TEX (131000 +/- 21000 cells/well). The thrombogenicity of the ECM was reduced by a mild glutaraldehyde treatment, performed prior to seeding the cells, accompanied by a 40-50% reduced endothelial cell growth. These results indicate that ECM provides a suitable biolayer for endothelial cell adhesion, growth and differentiation. It contains both adhesion glycoproteins (fibronectin, laminin, collagen) and endothelial cell growth factors (basic fibroblast growth factor) that support adhesion and normal growth of suboptimal concentrations of endothelial cells. We suggest that the presence in ECM of both adhesive macromolecules and potent endothelial cell growth promoting factors will make the ECM a promising substrate for vascular grafts.
天然血管与人工血管的明显区别在于其存在内皮细胞内衬，该内衬不仅具有抗血栓形成的特性，还具备修复和更新能力。本研究描述了一种在植入前用均匀的天然产生的内皮下细胞外基质（ECM）包被血管移植物的方法。将牛主动脉内皮细胞以低密度接种在未处理的戈尔特斯（GORE-TEX）、预先用ECM包被的戈尔特斯或先用纤连蛋白包被然后再用ECM包被的戈尔特斯上。细胞在无成纤维细胞生长因子（FGF）的情况下培养，并在培养7天后测定细胞数量。在未处理的戈尔特斯上观察到生长不佳（6030±1030个细胞/孔），在ECM包被的戈尔特斯上观察到生长改善了10倍（59900±1560个细胞/孔），而在用纤连蛋白-ECM预先包被的戈尔特斯上获得了最佳结果（131000±21000个细胞/孔）。在接种细胞之前进行轻度戊二醛处理可降低ECM的血栓形成性，但同时内皮细胞生长减少40 - 50%。这些结果表明，ECM为内皮细胞的黏附、生长和分化提供了合适的生物层。它既包含黏附糖蛋白（纤连蛋白、层粘连蛋白、胶原蛋白），也包含支持内皮细胞在次优浓度下黏附和正常生长的内皮细胞生长因子（碱性成纤维细胞生长因子）。我们认为，ECM中同时存在黏附性大分子和强大的内皮细胞生长促进因子将使ECM成为血管移植物的一种有前景的基质。
