Heparin and hyperbaric oxygenation in enteric autonomic neuron preservation for transplant.

BACKGROUND: Previous studies have suggested that the addition of heparin to a preservation solution attenuated the autonomic dysfunction observed in rat jejunum and in addition that hypothermic hyperbaric oxygenation may play a role as a preservation technique. However, these studies did not address the lesion indices of the autonomic enteric neurons. We sought to investigate whether the autonomic enteric neurons are injured during cold ischemic preservation and whether administration of heparin or hyperbaric oxygenation prevents this lesion. METHODS: Jejunal segments (2 cm; n = 20) of Wistar rats (12-16 weeks old) were maintained in lactated Ringer's solution without or with heparin (H- and H+, respectively) at 4 degrees C under normobaric conditions. Other jejunal segments (n = 10) were maintained at 4 degrees C in H- under hyperbaric oxygenation conditions (HBO). After preservation for 12 hours, H-, H+, and HBO preparations fixed in 10% formaldehyde were stained with hematoxylin and eosin. The lesion indices were expressed as the mean number of affected neurons (karyorhexis, nuclear dislocation, cytoplasmic vacuolisation) per 100 neurons present in intramural ganglia. Statistical analysis was performed using the Mann-Whitney test (P < .05). RESULTS: The histologic studies showed that enteric autonomic neurons were damaged in H- jejunal segments. The lesion indices observed were: karyorhexis 90/100, nuclear dislocation 85/100, and cytoplasmic vacuolization 82/100. The autonomic neurons in H+ and HBO segments seemed to be normal and significantly well-preserved (P < .001). CONCLUSION: Hypothermic hyperbaric oxygenation and heparin prevented lesions in cold ischemic preservation of enteric autonomic neurons.

Use of hyperbaric oxygenation in small bowel preservation for transplant.

INTRODUCTION: The aim of this work was to study the effects of hyperbaric oxygenation as a preservation technique for small bowel transplantation. METHODS: Twenty 2-month-old male Wistar rats weighting 250 g were divided into two groups: group A (n = 10) in which the small bowel was preserved for 12 hours, and group B (n = 10) in which the small bowel was preserved for 24 hours. After vascular and intraluminal perfusion, 3-cm segments were maintained in Ringer's solution at temperatures between 2 degrees C to 4 degrees C and in normobaric O2 conditions (groups A1, B1) or conditioned in an hyperbaric O2 metal chamber (100% oxygen at 5.5 absolute atmospheres) (groups A2, B2). After this preservation time, we studied intestinal tissue injury and morphometric analysis of the villi. RESULTS: Mucosal injury was significantly greater among group A1 compared to group A2 animals. The grade of the lesions was greater among group B1 compared to group B2 animals. Group A1 showed no difference from Group B1. For lesion grade, the range was smaller in group A2 and group B2 animals. The villi height was significantly smaller in groups A1 and B1 compared to the other groups; whereas it was higher in group A2 as compared with B2. CONCLUSION: Hyperbaric oxygenation may play a role as a preservation technique. Further research is necessary.

Apoptosis and nuclear proliferation in rat small bowel submitted to hypothermic hyperbaric oxygenation for preservation.

UNLABELLED: This study was conducted to assess apoptosis and nuclear proliferation in rat small bowel submitted to hypothermic hyperbaric oxygenation for preservation. METHODS: Twenty two-month-old, male Wistar rats, weighing 250 g were divided into two groups: group I (n = 10), in which the small bowel was preserved for 12 hours, and group II (n = 10) in which the small bowel was preserved for 24 hours. After vascular and intraluminal perfusion, 3-cm segments were maintained in Ringer's solution at 2 degrees to 4 degrees C under normobaric conditions (groups Ia and IIa) or conditioned in a small hyperbaric metal chamber with 100% oxygen at 5.5 absolute atmospheres (groups Ib and IIb). After 12 or 24 hours, apoptotic and mitotic indices were evaluated by immunohistochemical methods. RESULTS: The apoptotic index was significantly higher in small bowel segments in groups Ia and IIa compared with groups Ib and IIb. The mitotic index was significantly higher among group IIb. CONCLUSION: Hypothermic hyperbaric oxygenation reduced intestinal epithelial apoptosis and increased nuclear proliferation during rat small bowel preservation.

A novel system for organ and tissues preservation: the refrigerating hyperbaric chamber.

UNLABELLED: This study was designed to investigate the feasibility of building a simple and inexpensive device to preserve organs or tissues in hyperbaric and hypothermic conditions. METHODS: The device was built on a 40-cm wide, 28-cm long, and 23-cm deep stainless steel chassis. The pressure vessel was built by a 7.8-cm bore stainless steel cylinder put inside another 12-cm cylinder welded together and closed by a steel plate on the top and bottom. The inferior plate was welded, and the superior one was fixed by manual clasp nut. The cooling system is made up of air compressor, condenser, expansion area, and cooling worm that is located between the cylinders. The temperature-controlling device is a computer processor contained in an integrated-circuit chip, with a on-off system to maintain the chamber temperature between 2 degrees to 4 degrees C. The compression of the chamber is performed by lateral coupling with the oxygen cylinder and is maintained at 5.5 absolute atmospheres and controlled by air pressure gauge. The maximal work pressure was evaluated by spreadsheet. Temperature or pressure changes were evaluated by 12- and 24-hour assays. RESULTS: The maximal work pressure permitted was 6.5 absolute atmospheres. Thus, the container was free from danger. The temperature inside the chamber was kept between 2 degrees and 4 degrees C. The production costs of the prototype was US$1000. DISCUSSION: The manufacture of the refrigerating hyperbaric chamber is viable, simple, and inexpensive.