## 2 Results and Discussion
### Assessment of Anti-Inflammatory Properties  
The anti-inflammatory properties of the developed hydrogels with CBD and TER were assessed using three in vitro assays. As part of these studies, the effect of the studied DDSs on protein (bovine serum albumin) denaturation and the activity of proteinase and lipoxygenase were assessed. The results showed that both CBD and TER hydrogels have anti-inflammatory activity. In contrast, the best properties were obtained when both compounds were loaded simultaneously into the structures of the developed hydrogel carriers. For this hydrogel, an inhibition of protein denaturation of more than 46.0% was achieved when the 5.0% hydrogel concentration was used (Figure 12). Similar results were observed in an evaluation of the inhibition of the activity of two tested enzymes, proteinase and lipoxygenase, for which the inhibition of activity in the case of 5.0% H-CBD-TER was noted by 49.0 and 53.0%, respectively. Hydrogels containing TER and CBD individually also showed anti-inflammatory action; however, the results of inhibition of denaturation and enzyme activity were slightly lower (Figures 13 and 14).  
Figure 12: Inhibition of protein denaturation by tested hydrogels. *** \(p<0.0001\), * \(p=0.0414\) versus the control.  
Many skin conditions are related to inflammation [54]. As a result, developing DDSs containing active ingredients that control the pro-inflammatory response is extremely desirable. The results presented above illustrate the anti-inflammatory capabilities of CBD- and TER-containing hydrogels, which were confirmed in all three tests. Proteases are enzymes that can promote inflammation, mainly by regulating the expression and activity of various pro-inflammatory cytokines or chemokines. Hence, the ability of CBD and TER to inhibit proteinase activity may play an essential role in the anti-inflammatory activity of the developed DDSs [55]. In addition, the possibility of inhibiting lipoxygenase activity by the analyzed DDSs may contribute to reducing the production of pro-inflammatory mediators such as leukotrienes and affect the generation of anti-inflammatory mediators, which are lipoxins [56]. Tissue protein denaturation is a marker of inflammation and induces an inflammatory reaction. Hence, the possibility of inhibiting protein denaturation by the  
Figure 14: Inhibition of lipoxygenase activity by tested hydrogels. *****\(p<0.0001\) versus the control.  
Figure 13: Inhibition of proteinase activity by tested hydrogels. *****\(p<0.0001\) versus the control.  
developed hydrogels shown in this study also confirms the anti-inflammatory properties of the studied biomaterials [57; 58]. The anti-inflammatory effect of the studied DDSs, in addition to inhibiting the activity of pro-inflammatory enzymes and protein denaturation, is related to the multidirectional action of CBD, which can lower the level of pro-inflammatory cytokines, inhibit proliferation and induce apoptosis of T lymphocytes, and significantly reduce migration and adhesion of immune cells [16; 59]. This phytocannabinoid can also inhibit the activation of pro-inflammatory cells, influence the synthesis of pro-inflammatory mediators, and reduce intracellular and mitochondrial oxidative stress [16; 59]. CBD can also reduce reactive lipid peroxidation products and proteins, preventing changes in cell membranes' physical properties and function [16].