# The Effects of _Anthyllis vulneraria_ Hydroalcoholic Leaf Extract as an Adjuvant in Wound Healing
## 4 Discussion
Additionally, on a rat wound model, an extract from _Eugenia pruniformis_, a plant rich in hyperoside, quercetin, and kaempfer, aided in reepithelialization and wound architecture when compared to the controls, which was quantified microscopically through collagen deposition [65]. Ferulic acid, another substance identified in _A. vulneraria_ leaves, exerts anti-inflammatory effects by decreasing the levels of interleukins 1B and 6 (IL-1B, IL-6), along with tumor necrosis factor \(\alpha\) (TNF-\(\alpha\)). It also inhibited NLRP3 inflammasome activation, a structure closely associated with innate immunity, which leads to the release and activation of various caspases and IL-1B [74]. In one in vivo study on a rat model of radiotherapy skin wounds, ferulic acid reduced inflammation and oxygen radical species, and improved skin blood flow, tissue reconstruction, and collagen deposition [68]. Based on macroscopic findings, the EG1 and EG2 creams showed beneficial effects for the healing process on day 5, while on day 7, only the EG1 cream demonstrated accelerated healing, probably due to different concentrations of polyphenols. Therefore, we could argue that _A. vulneraria_ extract has a beneficial effect on wound healing in the first and early second stages of healing by providing an anti-inflammatory and antioxidant response, as well as encouraging epithelelization. This is also confirmed by the increased levels of antioxidant enzymes and reduced levels of oxidative stress markers and IL-8 amounts detected in the skin samples. After the early healing stages, the effects of _A. vulneraria_ no longer exceed those of the NC group, suggesting that _A. vulneraria_ can be an adjuvant of early healing. The possible loss of effect of EG2 cream on day 7 compared to EG1 cream could be explained by the detrimental effects of polyphenols, if present in a great amount. It may be that high levels of polyphenols can interfere with fibroblast migration and have cytotoxic effects [20; 75; 76], or could diminish the anti-inflammatory response in the wound, a crucial step in healing. This information can serve as an argument for calculating and using optimal concentrations of polyphenols for healing, as other studies have also indicated [20; 40; 41]. Considering the tested oxidative stress markers, for GSH, GSH/GSSG, SOD, and GPX a higher value is indicative of a better healing process, as these enzymes have protective antioxidant effects. For MDA and GSSG, a lower level is indicative of the healing response, as these two are markers of oxidative stress. We concluded that the EG2 group showedthe best biochemical healing process, correlated with the high content of polyphenols including hyperoside, ferulic acid, \(p\)-coumaric acid, gentisic acid, epigallocatechin gallate, and protocatechic acid. The EG1 groups showed similar results. Although these presented a weaker antioxidant response, the results were still comparable to EG2 and sometimes even better than the PC. Regarding IL-8, this cytokine is an indicator of early inflammation and a potent chemoattractant for neutrophils, while also influencing key healing aspects such as angiogenesis and cell migration [77; 78; 79]. One previous study in burn patients determined that high levels of IL-8 correlated with increased inflammation, sepsis, multiorgan failure, and mortality [80]. IL-8 also induced the formation of neutrophil extracellular traps, which can lead to thrombosis and intravascular disseminated coagulation during sepsis, as another study showed [81]. Thus, high levels of IL-8 could be indicators of impaired healing and overall increased mortality and morbidity. CAT catalyzes the conversion of H\({}_{2}\)O\({}_{2}\) into H\({}_{2}\)O; similar values in samples of different groups suggest a possible high activity of SOD and GPX, antioxidant enzymes that work in tandem to neutralize oxidative stress by reducing hydrogen peroxide, superoxide radicals, and organic hydroperoxides, thereby preventing cellular damage and maintaining redox balance [54].