Low-Frequency Low-Intensity Ultrasounds Do Not Influence the Survival and Immune Functions of Cultured Keratinocytes and Dendritic Cells
Table 1
Effects of low-frequency US at 0.15–7 W/cm 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 , 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 MFI, 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/cm)
Apoptotic cells (%)
ICAM-1 (MFI)
HLA-DR (MFI)
64 integrin (MFI)
CXCL10 (pg/ cells)
CCL2 (pg/ cells)
CXCL8 (ng/ cells)
Phopho-ERK1/2 (F.I.)
IL-6 (pg/ cells)
TNF- (pg/ 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/cm)
Apoptotic cells (%)
CD1a (MFI)
CD40 (MFI)
CD83 (MFI)
CD80 (MFI)
HLA-DR (MFI)
CD86 (MFI)
IL-12 (pg/mL)
IL-6 (pg/mL)
TNF- (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.
