V. M. Baratta1, T. M. Gisinger1,2, M. J. Barahona1, J. Ollodart1, D. Mulligan1, J. P. Geibel1,3 1Yale University School Of Medicine,Department Of Surgery,New Haven, CT, USA 2Paracelsus Medical University,Department Of Medicine,Salzburg, SALZBURG, Austria 3Yale University School of Medicine,Department Of Cellular And Molecular Physiology,New Haven, CT, USA
Introduction: Intestinal failure is the inability to maintain nutritional autonomy due to dysfunction or loss of functional native intestine. In both pediatric and adult populations, a heterogenous list of disorders may lead to intestinal failure and necessitate either total parenteral nutrition (TPN) or intestinal transplantation. Unfortunately, intestinal transplantation remains technically challenging. One of the barriers to successful intestinal transplantation is the progression of ischemia from tissue harvest to implantation. Finding ways to abate small bowel ischemic injury will help prolong graft function and viability during and after procurement. In this study, we demonstrate how exposure of the small intestine to Penicillin G can protect from ischemic injury in a rat model.
Methods: Distal and middle small bowel segments were harvested and attached to an ex-vivo intestinal perfusion device. Each segment was maintained at 37°C and perfused both from the luminal and basolateral side. FITC-Inulin (fluorescein isothiocyanate-inulin), was used to assess the ischemic conditions of the intestinal grafts in real-time, as previously described. A decrease in fluorescence is reflective of the onset of cellular ischemic injury. The perfused bowel segments were bathed in a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), HEPES-Ringer solution. The extraluminal buffer solutions were bubbled with 100% nitrogen to mimic intestinal ischemia. The experimental bowel segments were exposed to intraluminal 5 mM Penicillin G, while the control segments were perfused without Penicillin G.
Results: Control (without Penicillin G) small bowel samples showed significantly greater ischemic injury than experimental segments (with Penicillin G), as reflected by differences in FITC-Inulin fluorescence concentrations (i.e. decreased ischemic injury) compared with experimental segments (47.37 ± 0.7288 μM FITC-Inulin vs. 35.76 ± 4.469 μM FITC-Inulin, p 0.0248, Figure 1).
Conclusion: Penicillin G-exposed small intestines are more resistant to ischemic injury than segments without antibiotic exposure. This was demonstrated by the drop in fluorescent concentrations over time, which reflects cellular fluid secretion and apoptosis. This finding may have numerous applications in the field of transplant surgery, such as prolonging organ viability following perfusion with Penicillin G.
