Buffered formalin

Colon macroscopically visible damage was examined blindly, immediately after resection using a 0-5 scale, which takes into account the area of inflammation and the presence or absence of ulcers [25 (link)]. Then, colon sections were collected for the histological examinations. The material was fixed in 4% buffered formalin (ChemPur, Poland), embedded in paraffin, cut into 4 μm thick slices using a rotary microtome (Sakura Accu-Cut SRM, Netherlands), and placed on glass slides. Preparations were stained using three methods: hematoxylin-eosin, Giemsa, and alcian blue for histological assessment of colonic damage, cell infiltration, and mucus content, respectively. Tissue sections were evaluated, and images were taken by standard light microscopy (Olympus CX41 microscope coupled to a DP20 camera, Olympus Soft Imaging Solutions GmbH–cell^D ver. 3.2, Olympus, Germany). Histological damage was assessed in a blinded fashion by the experienced pathologist using the scoring scales, based on the previously presented criteria [26 (link)]. In hematoxylin-eosin staining, the result was the sum of points (0-25) scored by evaluation of each criterion. In Giemsa staining, inflammatory cell infiltration was assessed using a 0-4-point scale. Specimens stained with alcian blue were assessed by description considering goblet cell number and size and amount of sialomucins.

ENX was purchased from Sanofi-Aventis, Gentilly, France. PS, Grade X, was purchased from Sigma-Aldrich, Darmstadt, Germany. Phosphate buffered saline (PBS) and trisodium citrate (≥99%) were purchased from Sigma-Aldrich, Darmstadt, Germany and isoflurane was purchased from Baxter, Unterschleißheim, Germany. Antifactor Xa and IIa assay kits were purchased from BioMedica Diagnostic, Windsor, NS, Canada. Lidocaine spray 10% was purchased from Egis-Pharmaceutical, Budapest, Hungary, and 10% buffered formalin was purchased from Chempur, Piekary Slaskie, Poland. HBC was synthesized as previously described [13 (link)].