Contribution of usage to endoscope working channel damage and bacterial contamination.

BACKGROUND: Biofilm formation has been shown to be associated with damaged areas of endoscope channels. It was hypothesized that the passage of instruments and brushes through endoscope channels during procedures and cleaning contributes to channel damage, bacterial attachment and biofilm formation. AIM: To compare surface roughness and bacterial attachment in used and new endoscope channels in vivo and in vitro. METHODS: Surface roughness of 10 clinically used (retired) and seven new colonoscope biopsy channels was analysed by a surface profiler. For the in-vitro study, a flexible endoscope biopsy forceps was passed repeatedly through a curved 3.0-mm-diameter Teflon tube 100, 200 and 500 times. Atomic force microscopy was used to determine the degree of inner surface damage. The number of Escherichia coli or Enterococcus faecium attached to the inner surface of the new Teflon tube and the tube with 500 forceps passes in 1 h at 37oC was determined by culture. RESULTS: The average surface roughness of the used biopsy channels was found to be 1.5 times greater than that of the new biopsy channels (P=0.03). Surface roughness of Teflon tubes with 100, 200 and 500 forceps passes was 1.05-, 1.12- and 3.2-fold (P=0.025) greater than the roughness of the new Teflon tubes, respectively. The number of E. coli and E. faecium attached to Teflon tubes with 500 forceps passes was 2.9-fold (P=0.021) and 4.3-fold (P=0.004) higher compared with the number of E. coli and E. faecium attached to the new Teflon tubes, respectively. CONCLUSION: An association was found between endoscope usage with damage to the biopsy channel and increased bacterial attachment.

Stable thrombus formation on irradiated microvascular endothelial cells under pulsatile flow: Pre-testing annexin V-thrombin conjugate for treatment of brain arteriovenous malformations.

BACKGROUND: Our goal is to develop a vascular targeting treatment for brain arteriovenous malformations (AVMs). Externalized phosphatidylserine has been established as a potential biomarker on the endothelium of irradiated AVM blood vessels. We hypothesize that phosphatidylserine could be selectively targeted after AVM radiosurgery with a ligand-directed vascular targeting agent to achieve localized thrombosis and rapid occlusion of pathological AVM vessels. OBJECTIVE: The study aim was to establish an in vitro parallel-plate flow chamber to test the efficacy of a pro-thrombotic conjugate targeting phosphatidylserine. METHODS: Conjugate was prepared by Lys-Lys cross-linking of thrombin with the phosphatidylserine-targeting ligand, annexin V. Cerebral microvascular endothelial cells were irradiated (5, 15, and 25 Gy) and after 1 or 3 days assembled in a parallel-plate flow chamber containing whole human blood and conjugate (1.25 or 2.5 µg/mL). Confocal microscopy was used to assess thrombus formation after flow via binding and aggregation of fluorescently-labelled platelets and fibrinogen. RESULTS AND CONCLUSIONS: The annexin V-thrombin conjugate induced rapid thrombosis (fibrin deposition) on irradiated endothelial cells under shear stress in the parallel-plate flow device. Unconjugated, non-targeting thrombin did not induce fibrin deposition. A synergistic interaction between radiation and conjugate dose was observed. Thrombosis was greatest at the highest combined doses of radiation (25 Gy) and conjugate (2.5 µg/mL). The parallel-plate flow system provides a rapid method to pre-test pro-thrombotic vascular targeting agents. These findings validate the translation of the annexin V-thrombin conjugate to pre-clinical studies.