Extracellular matrix ligands promote RPE attachment to inner Bruch’s membrane

Abstract

Purpose. To determine if resurfacing layers of human Bruch’s membrane with extracellular matrix ligands increases adult human retinal pigment epithelium (RPE) attachment. Methods. We removed successive layers of Bruch’s membrane sequentially by mechanical or enzymatic means to expose the RPE basal lamina, inner collagen layer, and elastin layer. Each layer was coated with extracellular matrix proteins (4 µg/cm 2 laminin, 4 µg/cm 2 vitronectin, 20 µg/cm 2 fibronectin, 20 µg/cm 2 collagen IV) for 2 hrs at 37°C. First passage adult human RPE (15 000 cells/6mm explant) were plated onto coated or uncoated Bruch’s membrane layers and RPE attachment was measured 18 hrs later. Results. The reattachment rate to basal lamina derived from younger (age < 50) donors was higher than older (age = 70) donors (56.2 ± 4.8 % versus 46.7 ± 0.7 % respectively; p < 0.05). Coating with laminin or collagen IV increased RPE attachment to older but not younger basal lamina. The reattachment rate to inner collagen derived from younger donors was higher than older donors (51.3 ± 2.7 % versus 37.4 ± 1.7 % respectively, p < 0.05). Coating with laminin, fibronectin or collagen IV increased RPE attachment to older but not younger inner collagen layer. Adding extracellular matrix ligands did not increase RPE attachment to elastin. Conclusions. RPE attachment is increased by adding ligands to the basal lamina or inner collagen layer but not to the elastin layer. This may be important if the elastin layer is exposed in AMD patients after submacular surgery.