Figure 1. MTD-induced Permeability changes in EA and HPAEC.
(A) EA cells were seeded onto ECIS cultureware (8W10E+). Various concentrations (×1/100, ×1/50, ×1/25) of MTD were applied to EA cells at time “0” and capacitance changes were determined as described previously [8]. (B) HPAEC were applied to ECIS cultureware as describe in 1A. The magnitude of responses is different from EA cells but MTD (×1/25) markedly increases permeability in HPAEC as well. Protease-treated MTD does not increase permeability. Proteases were always heat inactivated before application of MTD to HPAEC. MTD activity is unaffected by heat. Heat-treated protease itself (HBSS-protease), medium and HBSS have no effect on EC permeability. (C) MTD (×1/25), PMN (2×105/well) from healthy volunteers were applied to EA cells along with MTD or in medium alone (N = 3 for MTD and MTD+PMN, N = 2 for Medium). (D) EA cells treated with medium or unstimulated PMN. These are control condition that apply to all ECIS experiments with EA cells. To avoid repetition, we show the typical medium or PMN (2×105 cells/well) treated EA cell responses. For 1A, B, C: Values were collected every 160 sec at 64,000 Hz as recommended by manufacture. Data were collected from 2–3 wells and each well has 40 electrodes. Thus each data point represents mean and SD values from at least total of 80 electrodes. All experiments were repeated at least three times.