IADR Abstract Archives
LINOLEIC ACID INDUCES A GAP CLOSURE MIGRATION OF HUMAN ORAL KERATINOCYTES CELLS BY AN INCREASE OF ACTIVITY OF CONNEXIN-BASED HEMICHANNELS
Objectives: We evaluated the effect of LA on gap closure migration mediated by Cx HCs in human oral keratinocytes
Methods: Human keratinocytes cell line (OKF6) was used. The cells were maintained in regular culture conditions (37°C, 5%CO2) with Keratinocytes-SFM medium supplemented with CaCl2. Mediated mechanical stress (“scratch”), a wound healing assay was evaluated in the absence or presence of LA (100 µM). OKF6 cells in presence (2 – 24 h) of LA were used for PCR assays. Also, in presence of LA the Cx HCs was evaluated by dye uptake assay
Results: In control conditions after 24 h of wound healing assays OKF cells had 25% of gap closure, but 24 h after LA treatment these cells had 65% of gap closure. Then, this increase in gap closure migration by LA was not evidenced in the presence of lanthanum (La3+, 200 µM), a Cx HCs inhibitor. At least, this cell type expresses Cx43 and Panx1. The expression of Cx43 (mRNA) increased after LA treatment (24 h). Panx1 expression did not change with LA treatment. Dye uptake assay showed activity of connexin-based hemichannels in the presence of LA. This activity was blocked in presence of La3+.
Conclusions: LA increases the gap closure migration of human oral keratinocytes through an increase of connexin hemichannels activity.
Methods: Human keratinocytes cell line (OKF6) was used. The cells were maintained in regular culture conditions (37°C, 5%CO2) with Keratinocytes-SFM medium supplemented with CaCl2. Mediated mechanical stress (“scratch”), a wound healing assay was evaluated in the absence or presence of LA (100 µM). OKF6 cells in presence (2 – 24 h) of LA were used for PCR assays. Also, in presence of LA the Cx HCs was evaluated by dye uptake assay
Results: In control conditions after 24 h of wound healing assays OKF cells had 25% of gap closure, but 24 h after LA treatment these cells had 65% of gap closure. Then, this increase in gap closure migration by LA was not evidenced in the presence of lanthanum (La3+, 200 µM), a Cx HCs inhibitor. At least, this cell type expresses Cx43 and Panx1. The expression of Cx43 (mRNA) increased after LA treatment (24 h). Panx1 expression did not change with LA treatment. Dye uptake assay showed activity of connexin-based hemichannels in the presence of LA. This activity was blocked in presence of La3+.
Conclusions: LA increases the gap closure migration of human oral keratinocytes through an increase of connexin hemichannels activity.