Investigation of the effect of proline on wound healing in an in vitro wound model

In this thesis, the effects of the amino acid proline on wound healing in vitro were investigated. Proline is known to play important roles in processes such as collagen synthesis, cell proliferation, and tissue repair in biological systems. The effects of proline on cell viability and wound healing were investigated using the HaCaT cell line. Proline solutions at doses of 0.01 mg/mL, 0.05 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.4 mg/mL, 0.5 mg/mL, 1 mg/mL, and 1.5 mg/mL, prepared by dissolving in sterile purified water, were added to the cell culture medium at a 10% concentration. The effects of different doses of proline solutions on cell viability were analyzed by MTT assay at 24, 48, and 72 hours. According to the VHC control group created by adding water to the culture medium, MTT results showed that proline applications at doses of 0.01 mg/mL (p<0.0001), 0.05 mg/mL (p<0.0001), 0.2 mg/mL (p=0.0031), 0.4 mg/mL (p=0.0035), 0.5 mg/mL (p=0.0003), 1 mg/mL (p<0.0001) and 1.5 mg/mL (p=0.0001) significantly increased viability at the 48th hour, but at the 72nd hour, cell viability decreased significantly at the dose of 0.2 mg/mL (p=0.0122). Based on MTT results, doses of 0.01 mg/mL, 0.05 mg/mL, 0.4 mg/mL, and 1.5 mg/mL, which demonstrated significant viability effects, were used in an in vitro scratch wound model. Microscope images were taken from the same areas of the scratch wound at 0, 24, 48, and 72 hours. Wound closure areas in the images were analyzed with ImageJ. In the VHC group, only doses of 0.4 mg/mL (p=0.0431) at 48 hours and 0.01 mg/mL (p=0.0443) and 0.4 mg/mL (p=0.0136) at 72 hours demonstrated significant wound closure. Statistical analyses were performed using Graphpad Prism, with a p<0.05 level of significance. The findings indicate that proline can positively affect cell viability, depending on dose and time, and has the potential to promote wound healing.