TY - JOUR
T1 - Effects of negative pressures on epithelial tight junctions and migration in wound healing
AU - Hsu, Chih Chin
AU - Tsai, Wen Chung
AU - Chen, Carl Pai Chu
AU - Lu, Yun Mei
AU - Wang, Jong Shyan
PY - 2010/8
Y1 - 2010/8
N2 - Negative-pressure wound therapy has recently gained popularity in chronic wound care. This study attempted to explore effects of different negative pressures on epithelial migration in the woundhealing process. The electric cell-substrate impedance sensing (ECIS) technique was used to create a 5 × 10-4 cm2 wound in the Madin-Darby canine kidney (MDCK) and human keratinocyte (HaCaT) cells. The wounded cells were cultured in a negative pressure incubator at ambient pressure (AP) and negative pressures of 75 mmHg (NP75), 125 mmHg (NP125), and 175 mmHg (NP 175). The effective time (ET), complete wound healing time (T max), healing rate (Rheal), cell diameter, and wound area over time at different pressures were evaluated. Traditional wound-healing assays were prepared for fluorescent staining of cells viability, cell junction proteins, including ZO-1 and E-cadherin, and actins. Amount of cell junction proteins at AP and NP125 was also quantified. In MDCK cells, the ET (1.25 ± 0.27 h), Tmax (1.76 ±0.32 h), and R heal (2.94 ± 0.62 × 10-4 cm2/h) at NP125 were significantly (P < 0.01) different from those at three other pressure conditions. In HaCaT cells, the Tmax (7.34 ± 0.29 h) and Rheal (6.82 ± 0.26 × 10-5 cm2/h) at NP125 were significantly (P < 0.01) different from those at NP75. Prominent cell migration features were identified in cells at the specific negative pressure. Cell migration activities at different pressures can be documented with the real-time wound-healing measurement system. Negative pressure of 125 mmHg can help disassemble the cell junction to enhance epithelial migration and subsequently result in quick wound closure.
AB - Negative-pressure wound therapy has recently gained popularity in chronic wound care. This study attempted to explore effects of different negative pressures on epithelial migration in the woundhealing process. The electric cell-substrate impedance sensing (ECIS) technique was used to create a 5 × 10-4 cm2 wound in the Madin-Darby canine kidney (MDCK) and human keratinocyte (HaCaT) cells. The wounded cells were cultured in a negative pressure incubator at ambient pressure (AP) and negative pressures of 75 mmHg (NP75), 125 mmHg (NP125), and 175 mmHg (NP 175). The effective time (ET), complete wound healing time (T max), healing rate (Rheal), cell diameter, and wound area over time at different pressures were evaluated. Traditional wound-healing assays were prepared for fluorescent staining of cells viability, cell junction proteins, including ZO-1 and E-cadherin, and actins. Amount of cell junction proteins at AP and NP125 was also quantified. In MDCK cells, the ET (1.25 ± 0.27 h), Tmax (1.76 ±0.32 h), and R heal (2.94 ± 0.62 × 10-4 cm2/h) at NP125 were significantly (P < 0.01) different from those at three other pressure conditions. In HaCaT cells, the Tmax (7.34 ± 0.29 h) and Rheal (6.82 ± 0.26 × 10-5 cm2/h) at NP125 were significantly (P < 0.01) different from those at NP75. Prominent cell migration features were identified in cells at the specific negative pressure. Cell migration activities at different pressures can be documented with the real-time wound-healing measurement system. Negative pressure of 125 mmHg can help disassemble the cell junction to enhance epithelial migration and subsequently result in quick wound closure.
KW - Cell junctions
KW - Cell movement
KW - Electric impedance
KW - Negative-pressure wound therapy
UR - https://www.scopus.com/pages/publications/77955858693
U2 - 10.1152/ajpcell.00504.2009
DO - 10.1152/ajpcell.00504.2009
M3 - 文章
C2 - 20445172
AN - SCOPUS:77955858693
SN - 0363-6143
VL - 299
SP - C528-C534
JO - American Journal of Physiology - Cell Physiology
JF - American Journal of Physiology - Cell Physiology
IS - 2
ER -