Abstract
The efficient translation of various experimental findings is hampered by interspecies (or even inter-strain) differences that may often invalidate findings, mechanisms or therapeutic approaches. Intestinal preservation/reperfusion injury is a field where rodents were extensively used yet any eventual clinical application would require validation in large animal models. We compared the development of tissue injury in rat and porcine intestines undergoing static cold storage (SCS) in histidine-tryptophane-ketoglutarate (HTK) solution. Methods: Standard procurement and perfusion of the small intestine was performed in Sprague Dawley rats and Landrace pigs (N=7 in each group). At 8h, 14h and 24 hours’ tissue samples were obtained and assessed for tissue damage (Chiu/Park score), apoptosis (active caspase-3) and tissue expression of ZO-1, claudin-3 and villin (immunofluorescence). Results: Rats showed a moderate mucosal injury (median Chiu/Park grade 3) already after 8h while pigs showed normal or near-normal histology (median grade 1). Histology remained significantly superior in pigs after 14h and 24h (see figure). Rats had advanced mucosal injury while only mild/moderate mucosal injury (median grade 2) was noted in pigs after 24h of SCS. Apoptotic enterocytes were abundant in rat but not pig intestines after 14 hours of SCS. After 14 hours of SCS, immunofluorescence revealed the virtual absence of ZO-1 staining in the rat but not in porcine intestines where a strong signal was detected even after 24 hours of SCS. Claudin-3 expression gradually changed from reticular staining on the basolateral membrane to a combined (membrane & cytoplasmic) signal in both species (albeit faster in rats). Conclusions: Pig intestines are significantly more resilient to preservation injury than rat intestines and longer SCS should be considered to attain a visible histological injury. The sequential alterations of certain molecular targets could serve as surrogate injury markers.Figure
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