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The human amniotic basement membrane model was utilized to determine diffusion ratios of dextrans and beads according to size selectivity. Diffusion through both intact and denuded amnions was determined after 24 and 72 h. Four neutrally charged fluorescein isothiocyanate labeled dextrans, having molecular weights of 17,900, 42,000, 71,200 and 156,000, diffused through the amnion. The amnion functioned as a sieve in that the passage of dextrans was increasingly restricted as molecular weight increased. In contrast, uncharged latex microspheres (1.05 micron +/- 0.07 micron (SD] and fluorescent carboxylated microspheres (1.57 micron +/- 0.13 micron (SD] failed to pass through the amnion. Light and electron microscopy revealed no preformed channels through which the 1.05 micron microspheres could pass through the amnion. Statistical analysis of cross-sectional thickness of individual and similarly treated amnions (intact or denuded) showed a difference in thickness (P = 0.05).

The effect of butyric acid (BA) and all trans-retinoic acid (RA) on murine melanoma cells was investigated in vitro and in vivo. The in vitro assays included 3H-IdUR incorporation, adhesion, migration and invasion experiments. Butyric acid decreased 3H-IdUR cellular incorporation within 24 h and increased adhesion as measured by trypsin release of 3H-IdUR labelled cells from either polycarbonate (p.c.) or Matrigel-coated p.c. membranes. Migration and invasion rates after 72 h were quantified by scanning electron microscopy (SEM). The invasion barrier consisted of Matrigel-coated p.c. membranes. Butyric acid significantly enhanced migration and invasion of B16a cells, while RA significantly decreased migration and invasion of B16a and K-1735 cells. Subcutaneous administration of either BA or RA pellets significantly decreased the number of lung nodules in the experimental metastatic assay. The experimental metastatic assay is defined as a tail vein inoculation protocol followed by subsequent lung evaluation.