Pınar Naile ÖĞÜTEN, Selin ENGÜR ÖZTÜRK, Miriş DİKMEN

Investigation of the Effectiveness of Cl-Amidine on Wound Healing: An In Vitro Study

Objective: Peptidylarginine deiminases (PADs) are enzymes converting the arginine to citrulline. They play a role in embryogenesis and cell signaling activities. But excessive or dysregulated PAD levels were determined to be associated with disorders and to increase in many diseases. It has been shown that Chloramidine (Cl-amidine) used as a PAD inhibitor suppresses increased PAD activity and shows anti-cancer, anti-inflammatory and antioxidant activities. Anti-inflammatory and antioxidant properties play an important role in wound healing. In this study, the possible efficacy of Cl-amidine on wound healing in the keratinocyte cell line was investigated by considering these parameters. Methods: Cell proliferation evaluations of Cl-amidine concentrations (500, 125, 31.25 and 7.81 µM) determined according to the results of MTT method on HaCaT keratinocyte cells were performed using Real-Time Cell Analysis System (RTCA DP). COL1A1 mRNA expression levels were analyzed by RT (Real Time)-PCR (Polymerase Chain Reaction) method at the concentrations where proliferation was achieved (125, 31.25 µM). Migration effects of Cl-amidine on cells were evaluated by performing scratch analysis. MTT results were statistically analyzed with one-way ANOVA and Tukey test, and p<0.05 was accepted as significant. RTCA DP and RT-PCR results were evaluated using device software programs. Results: In the study, it was found that certain concentrations of Cl-amidine had a proliferative effect on HaCaT keratinocyte cells. It was determined that Cl-amidine increased the amount of type 1 collagen, which is an important parameter for wound healing, by RT-PCR method. In addition, according to scratch analysis, it was detected that it positively affected cell migration in relation to wound closure. Conclusion: This research shows that Cl-amidine may have a significant potential for wound healing.

Keywords:

Peptidyl arginine deiminase (PAD) Cl-Amidine, Wound healing, HaCat keratinocyte cell line,

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