Murray H. Kown, MDa, Atsushi Yamaguchi, MDa, Christina L

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L -arginine polymers inhibit the development of vein graft neointimal hyperplasia Murray H. Kown, MDa, Atsushi Yamaguchi, MDa, Christina L. Jahncke, a, Douglas Miniati, MDa, Seiichiro Murata, MDa, Jurg Grunenfelder, MDa, Mark L. Koransky, MDa, Jonathan B. Rothbard, PhDb, Robert C. Robbins, MDa The Journal of Thoracic and Cardiovascular Surgery Volume 121, Issue 5, Pages 971-980 (May 2001) DOI: 10.1067/mtc.2001.112532 Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Representative figures of biotinylated L -arginine polymer and PBS-treated rabbit external jugular veins. Vessel lumina are on the right side of each photomicrograph. Nuclei are visible as light green areas , whereas dark brown regions signify areas of L -arginine polymer uptake. There is minimal biotin staining apparent in either PBS or R5 groups in contrast to the darkened nuclei and cytoplasm of R7 and R9 categories. (Original magnification, 400×.) The Journal of Thoracic and Cardiovascular Surgery 2001 121, 971-980DOI: (10.1067/mtc.2001.112532) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Representative figures of biotinylated L -arginine polymer and PBS-treated human saphenous veins. As is the case with rabbit veins, there appears to be little or no uptake of the polymers in either the PBS or R5 groups, whereas considerable expression of biotin is noted in the intima and media of vessels in both the R7 and R9 groups. (Original magnification, 400×.) The Journal of Thoracic and Cardiovascular Surgery 2001 121, 971-980DOI: (10.1067/mtc.2001.112532) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 Quantitative measure of translocation efficiencies on the basis of the number of biotin-staining nuclei (brown) divided by the number of total nuclei (brown plus green) in human saphenous vein samples. Each bar represents the average of 3 random high-power field readings of a cross-section of a single vessel. One hundred micromolar concentration appears to translocate with the highest efficiencies. Average translocation of the PBS and R5 group was significantly lower than that of R7 and R9 (P <.001, Student t test). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 971-980DOI: (10.1067/mtc.2001.112532) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 NO production as measured by nitrite (NO2-) levels. Pretreated venous segments are incubated in culture medium containing LPS (100 μg/mL) and IFN-γ (200 μg/mL) for 48 hours. Nitrite levels are measured directly from culture medium by means of Griess reaction and standardized to weights of venous segments. There is a length-dependent increase in nitrite levels with L -arginine polymer treatment. *P =.033 versus R5 100 μmol/L and P =.085 versus PBS (Bonferonni-corrected value). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 971-980DOI: (10.1067/mtc.2001.112532) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 A , Low-magnification view (100×) of the concentric nature of neointimal hyperplasia visible at 28 days in vein grafts stained with hematoxylin. Arrows delineate the neointimal layer. B , Higher magnification view (400×) of low, medium, and high degrees of neointimal hyperplasia. Arrows depict the border between medial and neointimal layers. The Journal of Thoracic and Cardiovascular Surgery 2001 121, 971-980DOI: (10.1067/mtc.2001.112532) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 6 I/M ratio of treatment and control groups of 28-day-old vein interposition grafts. I/M is lowest in both 100 μmol/L concentrations of R7 and R9. *P <.001 verus PBS, R5 10 μmol/L, and R5 100 μmol/L (Bonferonni-corrected value). The Journal of Thoracic and Cardiovascular Surgery 2001 121, 971-980DOI: (10.1067/mtc.2001.112532) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions