# The Effects of _Anthyllis vulneraria_ Hydroalcoholic Leaf Extract as an Adjuvant in Wound Healing
## 4 Discussion
From a biochemical perspective, hyperoside results from a highly efficient metabolic pathway involving the glycosylation of quercetin by UDP-galactosidaseyltransferase, a reaction that enhances its solubility, stability, and accumulation in vacuoles. Finally, the elevated concentration detected in our study is supported by a validated LC-MS/MS method using a high-purity analytical standard, ensuring both specificity and reliability. However, the absence of certain flavonoids, such as rutin, which has been widely reported in other studies [22; 24], suggests potential variations due to environmental factors, extraction methods, or plant physiology, as well as due to the controlled growth of _A. vulneraria_ specimens in non-native conditions as opposed to wild-type specimens, which were used in the other studies. More studies are needed to determine the origin of these discrepancies, as well as to determine the possible in vitro and in vivo interactions between the active substances of the extract. Interestingly, previous reports have rarely documented the presence of epigallocatechin gallate in _A. vulneraria_ leaves, thereby making our findings novel. The leaf extract, despite its modest overall polyphenol content, still contains quantifiable amounts of gentisic acid and epigallocatechin derivatives, which may provide anti-inflammatory benefits. The presence of catechins in the leaf extract suggests a possible role in plant defense mechanisms, as these compounds exhibit strong radical-scavenging activity and contribute to wound-healing effects through collagen stabilization [62]. The values we obtained for TPC and TFC can be compared to past findings in the literature. [PERSON] et al. obtained values of 82.86 mg gallic acid equivalent/g vegetal material for TPC, and 20.14 mg quercetin equivalent/g vegetal material for TPC [2], while the same group determined different values in another study, namely, 93.27 mg gallic acid equivalent/g vegetal material for TPC and 37.88 mg quercetin equivalent/g vegetal material for TFC [17]. When compared to our values (TPC 20.58 mg gallic acid equivalent/g vegetal material and TPC 31.03 mg quercetin equivalent/g vegetal material), we can see that while TFC has similar values, TPC is significantly diminished, possibly due to different growing and geographic conditions. Considering the antioxidant capacity of the leaf extract, the results we obtained can be compared to other data in the literature. [PERSON] et al. obtained values of 0.026 mM Trolox/g vegetal product for DPPH [63]. [PERSON] et al. obtained an ABTS value of 0.44 mM Trolox/g vegetal product [64], while [PERSON] et al. obtained an ABTS value of 0.82 mM Trolox/g vegetal product, while also finding a higher DPPH value of 0.33 mM Trolox/g vegetal product [2]; this is compared to our values for ABTS of 0.693 mM Trolox/g vegetal product and DPPH of 0.15 mM Trolox/g vegetal product. Overall, our results confirm the antioxidant capacity of the _A. vulneraria_ leaves. Other studies have found that hyperoside [65], ferulic acid [66] and _p_-comamic acid [67] have promising antioxidant, anti-inflammatory, and antimicrobial properties, while ferulic acid [66; 68], gentisic acid [69], and protocatechuic acid [70; 71] have shown good repepthelization and collagen-deposition capabilities, including in in vivo studies. Hyperoside, a common flavonoid found in numerous plants, was found to inhibit the formation of several key inflammatory factors. An in silico study showed that hyperoside has the capacity to bind to substances, such as nuclear factor-kB (NF-kB) and interleukins 17 and 36, thus inactivating their activity [72]. An in vivo/in vitro mouse model study found that hyperoside inhibits TNF-\(\alpha\), IL-6 and the high-mobility group box 1 (HMGB1), a nuclear protein closely involved in the innate immune response that leads to the activation of NF-kB [73].