Artificial matrix barriers: A diffusion study utilizing dextrans and microspheres

Abstract

Eight artificial matrices (AMs) were evaluated for the ability to restrict the passage of diffusion probes. Three AMs were composed exlusively of interstitial type I collagen (Col I) and differed from each other in thickness only. Four AMs consisted of reconstituted basement membrane (RBM) ‐coated polycarbonate filters (containing 10 μm diameter pores) and also only differed in thickness. One AM consisted of an uncoated 10 μm pore polycarbonate filter. The diffusion probes were uncharged fluorescein isothiocyanate‐labeled dextrans, having molecular weights of 17,900, 42,000, 71,200, and 148,900 and negatively charged latex microspheres, having diameters of 0.08, 0.30, and 0.95 μm. Probes were applied to the AMs, incubated for 72 hr at 37°C, and then analyzed spectrophoto‐metrically. Dextran passage was increasingly restricted for Col I matrices as either molecular weight or collagen thickness increased (range 7% to 0.7%). Thin RBM‐coated filters were more permeable to dextrans (range 100% to 30%) than Col I matrices. The diffusion rate of microspheres for Col I matrices (range 3.5% to 0) was similar to both thick and thin RBM‐coated filters (range 4% to 0). The uncoated filter permitted the most diffusion for both dextrans and microspheres (range 100% to 7%). These data demonstrate that the AMs presented in this study will allow direct observation of the degradative and migratory potential of cells in vitro as they interact with various extracellular matrices.