Raffinose improves 24-hour lung preservation in low potassium dextran glucose solution: a histologic and ultrastructural analysis Stefan Fischer, MD,

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Raffinose improves 24-hour lung preservation in low potassium dextran glucose solution: a histologic and ultrastructural analysis Stefan Fischer, MD, David Hopkinson, Mingyao Liu, MD, Alexandra A MacLean, MD, Vernon Edwards, MSc, Ernest Cutz, Shaf Keshavjee The Annals of Thoracic Surgery Volume 71, Issue 4, Pages 1140-1145 (April 2001) DOI: 10.1016/S0003-4975(01)02426-2

Fig 1 Changes in lung weight (postpreservation edema) after 24 hours of cold ischemic preservation in low potassium dextran (LPD) or raffinose-modified LPD (R-LPD). The correlation coefficient for the LPD regression line was 0.61 (p = 0.073) and for the R-LPD regression line, 0.98 (p = 0.004). The scatterplot demonstrates that the addition of raffinose to LPD contributes significantly to reducing weight gain during preservation as compared with LPD alone. Moreover, the effect increases with initial lung weight. The Annals of Thoracic Surgery 2001 71, 1140-1145DOI: (10.1016/S0003-4975(01)02426-2)

Fig 2 Proportions of dead cells in lungs after 24 hours of cold ischemic preservation in low potassium dextran (LPD) or raffinose-modified LPD (R-LPD) and in control lungs. Data are expressed as mean ± standard deviation. The numbers of dead cells are expressed as a percentage of total cells. However, the R-LPD group showed significantly lower numbers of dead cells (14% ± 1.4% of total lung cells) than lungs in the LPD group (29% ± 0.3%; p < 0.001). The R-LPD lungs also contained a significantly higher percentage of dead cells than control lungs with 0.2% ± 5% (p < 0.001). (One-way analysis of variance, ∗p < 0.001 among all groups.) The Annals of Thoracic Surgery 2001 71, 1140-1145DOI: (10.1016/S0003-4975(01)02426-2)

Fig 3 Representative images of lung tissue sections after using the transvascular trypan blue flush and an eosin counterstain. Dead cells lose their ability to actively exclude trypan blue and consequently stain dark blue. Each dark blue dot represents a dead cell. Control lungs (A) showed none to very little cell death, whereas lungs flushed with and stored in raffinose-modified low potassium dextran solution for 24 hours (B) demonstrated moderate cell death and lungs that were preserved with low potassium dextran solution only (C) showed high numbers of dead cells. The Annals of Thoracic Surgery 2001 71, 1140-1145DOI: (10.1016/S0003-4975(01)02426-2)

Fig 4 Representative medium-power electron micrograph from a control lung that was flushed with low potassium dextran solution without cold storage. Note the overall good preservation of the cells, including the type II pneumocyte (P2) containing multilamellar surfactant bodies (S). Capillaries (c) within the interstitium are normal and show intact endothelial lining (arrows) and well-preserved tight junctions (arrowheads). (×7,450.) The Annals of Thoracic Surgery 2001 71, 1140-1145DOI: (10.1016/S0003-4975(01)02426-2)

Fig 5 Representative electron micrographs from the lungs flushed with low potassium dextran solution and stored for 24 hours. (A) Low-power view demonstrates portions of several alveolar spaces and the lining alveolar cells. Note the cellular debris (arrowheads) and macrophage (M) within the alveolar spaces. Type I and type II pneumocytes are less distinguishable. The interstitium is of variable thickness, shows edema (∗), and contains necrotic cellular debris (D). Capillaries within this interstitium are poorly defined. (×4,630.) (B) High-power view shows an apoptotic cell (A) within an alveolar capillary. Note that this capillary is distorted and contains edema and cell debris (arrowheads). The type I pneumocytes (P1) lining the alveolar space are swollen and edematous in appearance, representing severe cellular injury. (×13,440.) The Annals of Thoracic Surgery 2001 71, 1140-1145DOI: (10.1016/S0003-4975(01)02426-2)

Fig 6 Representative electron micrographs from the lungs flushed with low potassium dextran solution containing raffinose and stored for 24 hours. (A) Low-power electron micrograph demonstrates the overall good preservation of the lung architecture and ultrastructure. Alveolar spaces are lined with a usual complement of type I and type II pneumocytes (P1, P2). The interstitium looks similar to the interstitium of control lungs and contains well-defined capillaries (c) with intact cell junctions (arrowheads). Note the absence of alveolar macrophages, apoptotic, or necrotic cells. (×3,850.) (B) High-power view illustrating the well-preserved ultrastructure of type II pneumocytes (P2) with multilamellar surfactant bodies (S). Alveolar capillaries (c) are within normal limits. The interstitium (I) shows no abnormality. (×14,560.) The Annals of Thoracic Surgery 2001 71, 1140-1145DOI: (10.1016/S0003-4975(01)02426-2)