Clinical, histopathological and serological investigation of the effects of ankaferd blood stopper® in the prevention of post-operative intra-abdominal adhesions: Experimental research on rats

Post-operative intra-abdominal adhesions are an important post-operative problem in surgery and can lead to serious complications which appear in the form of ileus, chronic pelvic pain, and infertility. In this study, we aimed to use and evaluate the effects of Ankaferd Blood Stopper® (ABS) for the prevention of post-operative intra-abdominal adhesions. ABS is an anti-bleeding plant extract obtained from Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum plants. In addition to its local bleeding stopping effect, the extract also has bacteriostatic and antibacterial properties. A total of 120 rats were divided into three operation groups (laparotomy group, laparotomy + cecal abrasion group, laparotomy + cornu suture group). Each treatment group was divided into four subgroups according to the applied solution: the 0.9% NaCl group, the ABS group, the group to which no substance was applied, and the Icodextrin (4%) group. A 3 cm skin incision was made on the ventral abdominal wall of each rat and the abdomen was entered. In the laparotomy group, the selected solutions were administered to all the rats without any procedure to the abdominal organs. In the laparotomy + cecal abrasion group, petechial bleeding was initiated by abrading the antimesenteric border of the cecum and the abdominal wall to which it contacts with standard strength gauze 20 times. Respective solutions were then applied to each subgroup. In the laparotomy + cornu suture group, the right cornu uteri of each subject was incised 1 cm cranially from the corpus uteri and were sutured with simple continuous suture. Before the abdomen was closed, respective solutions were applied into the abdomen of each subgroup. Rats were euthanized 21 days after the procedure. Intra-abdominal adhesions were evaluated quantitatively by previously decribed criteria. Biopsies were taken from the damaged surface and the attachment point of adhesion for histopathological examination. All slides were stained with hematoxylin-eosin and were evaluated for fibroblastic activity in tissues, inflammatory cells, and vascular proliferation. For statistical analysis, comparisons among all groups were expressed as microscopic, macroscopic findings and median (minimum-maximum) ELISA data. Kruskal-Wallis analysis of variance was used for microscopic and macroscopic comparison of ELISA data and adhesion scores of TIMP-1, MMP-9, IL-6, TNF-α, TGF-β among the groups. Dunn-Bonferroni test was used for post-hoc comparisons. The relationships between the variables were analyzed using the Spearman rank correlation coefficient. In statistical analyzes, p=0.05 was taken as the level of significance. At the end of the evaluations, a negative significant difference was found between the ABS and other groups for macroscopic evaluation scores. In terms of the microscopic evaluation of the histopathological examination, a negative significant difference was found between ABS and other groups. Ankaferd Blood Stopper® had no effect in preventing post-operative intra-abdominal adhesions, and also caused organ damage. Further studies are needed to find the mechanisms of adhesion formation and effective preventative agents.