Table 1.
Effects of low-frequency US at 0.15–7 W/cm2 intensity range on cultured keratinocytes and DC. Keratinocyte and DC cultures were treated with 42 KHz US at the indicated intensities for 7 minutes (1 cycle). Apoptotic cells were determined by annexin V/PI staining and FACS analysis. Apoptotic cell percentage (%) includes PI+, Ann V/PI+, as well as Ann V+ subpopulations. The expression of surface markers was revealed by performing staining with specific Abs and FACS analysis. Membrane molecule positivity is expressed as ΔMFI, which represents the mean fluorescence intensity of samples stained with specific Abs subtracted of the fluorescence of Ig-stained controls. Supernatants from keratinocyte and DC cultures were tested by ELISA to measure chemokine and cytokine contents. Phospho-ERK1/2 were revealed by Western blotting on lysates obtained from US-treated keratinocytes. Phopho-ERK1/2 amounts were calculated by densitometry and are expressed as fold induction (F.I.) in experimental relative to untreated samples, to which were given a value of 1.
| Keratinocytes | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| US intensity (W/cm2) | Apoptotic cells (%) | ICAM-1 (ΔMFI) | HLA-DR (ΔMFI) | α6β4 integrin (ΔMFI) | CXCL10 (pg/106 cells) | CCL2 (pg/106 cells) | CXCL8 (ng/106 cells) | Phopho-ERK1/2 (F.I.) | IL-6 (pg/106 cells) | TNF-α (pg/106 cells) |
| 0 (untreated) | 15.1 ± 1.9% | 5 ± 2 | 4 ± 2 | 61 ± 4 | 75 ± 9 | 61 ± 5 | 3 ± 0.02 | 1 | 0 | 71 ± 9 |
| 0.15 | 14.6 ± 2.3% | 5 ± 3 | 4 ± 1.5 | 62 ± 6 | 95 ± 10 | 70 ± 4 | 3.3 ± 0.17 | 0.9 ± 0.15 | 0 | 66 ± 8.5 |
| 0.66 | 16.05 ± 1.3% | 7 ± 2 | 5 ± 1.5 | 66 ± 3 | 100 ± 11 | 45 ± 7 | 3.4 ± 0.33 | 1.1 ± 0.08 | 0 | 83 ± 7 |
| 1.4 | 15.6 ± 2.2% | 8 ± 3 | 5 ± 3 | 62 ± 3.5 | 85 ± 9.5 | 77 ± 6 | 3.5 ± 0.32 | 1.1 ± 0.09 | 0 | 85 ± 9.5 |
| 2.3 | 16.6 ± 1.7% | 8 ± 1 | 3 ± 1 | 61 ± 5 | 77 ± 22 | 66 ± 9 | 3.70 ± 0.4 | 1.2 ± 0.2 | 0 | 69 ± 5 |
| 5 | 29.1 ± 2.65%§ | 16 ± 3.5# | 6 ± 0.5 | 75 ± 3§ | 81 ± 8.5 | 39 ± 6 | 6.66 ± 0.7# | 1.6 ± 0.07§ | 0 | 79 ± 6.5 |
| 7 | 53.5 ± 4.1%# | 18.5 ± 2# | 4 ± 1 | 81 ± 5§ | 110 ± 13 | 81 ± 11 | 7.5 ± 0.5* | 1.9 ± 0.08# | 0 | 64 ± 9 |
| DC | ||||||||||
| US intensity (W/cm2) | Apoptotic cells (%) | CD1a (ΔMFI) | CD40 (ΔMFI) | CD83 (ΔMFI) | CD80 (ΔMFI) | HLA-DR (ΔMFI) | CD86 (ΔMFI) | IL-12 (pg/mL) | IL-6 (pg/mL) | TNF-α (pg/mL) |
| 0 (untreated) | 19.1 ± 2.05% | 201 ± 7 | 19 ± 0.9 | 15 ± 0.7 | 30 ± 2.3 | 117 ± 6 | 201 ± 16 | 0 | 5 ± 0.1 | 105 ± 10 |
| 0.15 | 17.1 ± 2.5% | 199 ± 9 | 16 ± 1 | 13 ± 0.8 | 31 ± 2.2 | 112 ± 13 | 203 ± 18 | 0 | 7 ± 0.3 | 85 ± 9 |
| 0.66 | 20.2 ± 2.1% | 210 ± 11 | 18 ± 0.9 | 17 ± 0.9 | 40 ± 2.7 | 114 ± 9.5 | 197 ± 19 | 0 | 6 ± 1.6 | 80 ± 9.1 |
| 1.4 | 19.9 ± 2.4% | 200 ± 12 | 20 ± 1.1 | 11 ± 1.1 | 38 ± 2.5 | 110 ± 11.5 | 210 ± 11 | 0 | 5.5 ± 0.6 | 78 ± 7.1 |
| 2.3 | 23.1 ± 2.9% | 198 ± 15 | 23 ± 1.4 | 10 ± 1.1 | 31 ± 3 | 101 ± 8 | 204 ± 14 | 0 | 6.5 ± 0.6 | 125 ± 11 |
| 5 | 37.1 ± 2.8%§ | 170 ± 13 | 35 ± 2.1§ | 16 ± 0.9 | 37 ± 3.1 | 100 ± 16 | 201 ± 19 | 0 | 2 ± 0.9 | 78 ± 9.1 |
| 7 | 61.3 ± 3.8%# | 180 ± 9 | 37 ± 6§ | 18 ± 0.7 | 39 ± 3.1 | 120 ± 9.7 | 173 ± 16 | 0 | 10 ± 1.1 | 92 ± 8.1 |
§P < 0.05, versus untreated cells, #P < 0.01, versus untreated cells, *P < 0.001, versus untreated cells.