## 4 Discussion
Therefore, it plays a role as an \"alarmin\", further enhancing the inflammatory response [21; 22]. In contrast to our results, the serum level of IL-1\(\alpha\) did not differ between pediatric patients with CD and UC in the study conducted by [PERSON] et al. [8]. At the same time, in [PERSON] et al. 's study, the CCL2 serum profile was different between CD and UC pediatric groups, while no such difference was noted in our study. The observed inconsistency between our study and [PERSON] et al. 's may be related to the age of the patients, as [PERSON] et al. included only patients below 17 years old, while the mean age of patients included in our study was 32 in the CD group and 33 in the UC group. Moreover, in [PERSON] et al. 's [8] study, the activity of the disease was assessed with pediatric scales, and patients included in the research had to be in remission for at least 3 months, while patients in our study presented active disease. Therefore, the differences in the patients' inclusion criteria may be responsible for the observed differences between our study and [PERSON] et al. 's [8] study. The differences identified in our study in IL-1\(\alpha\) serum profile between CD and UC patients might be related to the greater role of that cytokine in the pathogenesis of CD among adults; however, this hypothesis should be further confirmed. The results of our study indicated, however, a valid role of IL-1\(\alpha\) in the diagnosis of IBD, as this parameter presented a moderate specificity and sensitivity in differentiating [PERSON]'s disease from ulcerative colitis. This study has some limitations, among which study size can be mentioned. Unfortunately, we were not able to confirm our results by comparing them with an independent cohort or \"in silico\". Expanding the number of patients included in study groups will enable a more precise evaluation of the specificity and sensitivity of the analyzed biomarkers, as well as allowing for the establishment of cut-off values of these biomarkers for differentiating [PERSON]'s disease from ulcerative colitis with higher precision. Considering the role of analyzed biomarkers in the inflammatory process, their serum profile may also be elevated in other inflammatory diseases. Therefore, further studies, including patients with different inflammatory diseases, are needed to elucidate the specificity of these biomarkers towards IBD. The performed ROC curve analysis of serum calprotectin and proinflammatory mediators revealed that E-selectin, P-selectin, IL-1\(\alpha\), and IL-12p70 presented a better specificity and sensitivity in differentiating CD from UC than serum calprotectin. However, pairing each parameter with the serum calprotectin level significantly increased the sensitivity and specificity of the diagnosis for both CD and UC. Therefore, simultaneous measurements of serum calprotectin with E-selectin, P-selectin, IL-1\(\alpha\), or IL-12p70 may present greater usefulness in the differential diagnosis of IBD than the measurement of each parameter individually. Unfortunately, our study did not include patients with unclassified colitis, preventing us from elucidating the usefulness of the analyzed proinflammatory mediators in the intermediate type of IBD. Thus, further research should include patients with unclassified colitis as another study group. One of the main advantages of our study is the simplicity of the methodology, particularly the multiplex test using the Luminex 200 flow cytometry system. This approach can be easily integrated into routine clinical practice because it is not time-consuming and allows the assessment of multiple parameters in a small sample volume.