Cytotoxicity and Apoptosis in BME Cells
| . | % Ethidium Homodimer Incorporation Above Control Levels . | % Acridine Orange-Labeled Cells . |
|---|---|---|
| Control | 7.18 ± 0.68 (n = 20) | |
| hPlg | ||
| 2.2 μg/mL | 0.67 ± 0.67 (n = 3) | ND |
| 11.0 μg/mL | ND | 7.55 ± 1.23 (n = 5) |
| 22.0 μg/mL | 0 (n = 4) | 5.99 ± 0.70 (n = 8) |
| 100 μg/mL | 2.3 (n = 1) | ND |
| rhTNF-α | ||
| 10.0 ng/mL | 27.05 ± 21.15 (n = 2) | 49.18 ± 9.51 (n = 6) |
| rmTNF-α | ||
| 1.0 ng/mL | 22.04 ± 6.50 (n = 5) | ND |
| 10.0 ng/mL | 30.79 ± 5.78 (n = 5) | ND |
| 1.0 μg/mL | ND | 77.68 ± 7.14 (n = 9) |
| hTGF-β1 | ||
| 1 ng/mL | 22.60 ± 18.40 (n = 2) | 13.95 ± 1.17 (n = 8) |
| LPS | ||
| 1 μg/mL | 4.90 ± 1.20 (n = 2) | ND |
| 10 μg/mL | 16.30 ± 10.70 (n = 2) | ND |
| ehAst | ||
| 1 μg/mL | 14.45 ± 3.35 (n = 4) | 12.32 ± 0.98 (n = 2) |
| 10 μg/mL | 21.61 ± 3.65 (n = 8) | 19.69 ± 2.49 (n = 8) |
| rhAst (CHO) | ||
| 1 μg/mL | 4.27 ± 2.13 (n = 3) | 13.80 ± 1.44 (n = 8) |
| 10 μg/mL | 8.00 ± 2.06 (n = 4) | ND |
| 50 μg/mL | 24.10 (n = 1) | 17.39 ± 3.54 (n = 6) |
| rhAst (P.p.) | ||
| 1 μg/mL | 4.05 ± 4.05 (n = 2) | ND |
| 10 μg/mL | 13.10 ± 6.50 (n = 2) | ND |
| rmAst (Bac.) | ||
| 1 μg/mL | 4.35 ± 0.85 (n = 2) | ND |
| 10 μg/mL | 12.25 ± 2.05 (n = 2) | 7.16 ± 0.47 (n = 8) |
| 25 μg/mL | ND | 38.75 ± 4.26 (n = 5) |
| . | % Ethidium Homodimer Incorporation Above Control Levels . | % Acridine Orange-Labeled Cells . |
|---|---|---|
| Control | 7.18 ± 0.68 (n = 20) | |
| hPlg | ||
| 2.2 μg/mL | 0.67 ± 0.67 (n = 3) | ND |
| 11.0 μg/mL | ND | 7.55 ± 1.23 (n = 5) |
| 22.0 μg/mL | 0 (n = 4) | 5.99 ± 0.70 (n = 8) |
| 100 μg/mL | 2.3 (n = 1) | ND |
| rhTNF-α | ||
| 10.0 ng/mL | 27.05 ± 21.15 (n = 2) | 49.18 ± 9.51 (n = 6) |
| rmTNF-α | ||
| 1.0 ng/mL | 22.04 ± 6.50 (n = 5) | ND |
| 10.0 ng/mL | 30.79 ± 5.78 (n = 5) | ND |
| 1.0 μg/mL | ND | 77.68 ± 7.14 (n = 9) |
| hTGF-β1 | ||
| 1 ng/mL | 22.60 ± 18.40 (n = 2) | 13.95 ± 1.17 (n = 8) |
| LPS | ||
| 1 μg/mL | 4.90 ± 1.20 (n = 2) | ND |
| 10 μg/mL | 16.30 ± 10.70 (n = 2) | ND |
| ehAst | ||
| 1 μg/mL | 14.45 ± 3.35 (n = 4) | 12.32 ± 0.98 (n = 2) |
| 10 μg/mL | 21.61 ± 3.65 (n = 8) | 19.69 ± 2.49 (n = 8) |
| rhAst (CHO) | ||
| 1 μg/mL | 4.27 ± 2.13 (n = 3) | 13.80 ± 1.44 (n = 8) |
| 10 μg/mL | 8.00 ± 2.06 (n = 4) | ND |
| 50 μg/mL | 24.10 (n = 1) | 17.39 ± 3.54 (n = 6) |
| rhAst (P.p.) | ||
| 1 μg/mL | 4.05 ± 4.05 (n = 2) | ND |
| 10 μg/mL | 13.10 ± 6.50 (n = 2) | ND |
| rmAst (Bac.) | ||
| 1 μg/mL | 4.35 ± 0.85 (n = 2) | ND |
| 10 μg/mL | 12.25 ± 2.05 (n = 2) | 7.16 ± 0.47 (n = 8) |
| 25 μg/mL | ND | 38.75 ± 4.26 (n = 5) |
Confluent monolayers of BME cells in 96-well plates were exposed to the agents indicated above, and cytotoxicity (ethidium homodimer incorporation) and apoptosis (acridine orange labeling) were determined after 72 and 48 hours, respectively. Values are the mean ± SEM. For ethidium homodimer incorporation, n is the number of experiments. For acridine orange labelling, n is the number of photographic fields analyzed; results are pooled from 4 separate experiments and have been normalized to the mean of the mean control values of the four experiments.
Abbreviations: ND, not determined; CHO, Chinese hamster ovary cells;P.p., P. pastoris; Bac., baculovirus.