Rupak Mukherjee, PhD, Juozas A. Zavadzkas, MD, Stuart M

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Targeted Myocardial Microinjections of a Biocomposite Material Reduces Infarct Expansion in Pigs Rupak Mukherjee, PhD, Juozas A. Zavadzkas, MD, Stuart M. Saunders, BS, Julie E. McLean, BS, Laura B. Jeffords, BS, Christy Beck, (HTL), BS, Robert E. Stroud, MS, Allyson M. Leone, BS, Christine N. Koval, BS, William T. Rivers, BS, Shubhayu Basu, PhD, Alexander Sheehy, MS, Gene Michal, PhD, Francis G. Spinale, MD, PhD The Annals of Thoracic Surgery Volume 86, Issue 4, Pages 1268-1276 (October 2008) DOI: 10.1016/j.athoracsur.2008.04.107 Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 (A) Intraoperative injection of the fibrin-alginate (Fib-Alg) composite was performed using a 2- × 2-cm template with injection sites arrayed at 0.5-cm intervals within the region of myocardial infarction (MI). (B) At necropsy, the Fib-Alg could be visualized as amorphous densities within the MI region. (LV = left ventricle.) The Annals of Thoracic Surgery 2008 86, 1268-1276DOI: (10.1016/j.athoracsur.2008.04.107) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 There were significant relationships between myocardial infarction (MI) size and left ventricular end-diastolic volume at 28 days after MI in the saline (circles; y = 3.06x + 40.74; r2 = 0.50, p = 0.03) and fibrin-alginate (Fib-Alg; squares) groups (y = 1.62x + 41.69; r2 = 0.44, p = 0.04). The Annals of Thoracic Surgery 2008 86, 1268-1276DOI: (10.1016/j.athoracsur.2008.04.107) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Representative low-power (top panels) and high-power (bottom panels) photomicrographs of hematoxylin and eosin–stained left ventricular sections at 28 days after either saline or fibrin-alginate (Fib-Alg) injection. Increased cellularity, which was focally distributed around amorphous densities, likely that of injected composite material (arrows with green borders), could be readily appreciated in sections from the Fib-Alg group. The edges of the amorphous bodies corresponding to the sites of the fibrin-alginate injections were well defined, suggesting that localized diffusion was minimal. High-power photomicrographs were imaged from the areas represented by the insets. In the Fib-Alg group, the presence of giant cells (arrows with red borders) was noted. Scale bars are 100 μm. The Annals of Thoracic Surgery 2008 86, 1268-1276DOI: (10.1016/j.athoracsur.2008.04.107) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 (A) Representative photomicrographs of immunostaining for lectin-positive capillaries from control left ventricular (top panels) and from the border and myocardial infarction (MI) regions (middle and bottom panels, respectively) after either saline or fibrin-alginate (Fib-Alg) injection within the MI region. Scale bar for all photomicrographs is 50 μm. (B). Summary of capillary density measurements. Compared with respective remote region values, the number of capillaries in the MI region was lower in the saline group, but remained similar in the Fib-Alg group. (C = p < 0.05 vs reference controls; R = p < 0.05 vs treatment-matched remote region). Data are presented as the mean and standard error of the mean. The Annals of Thoracic Surgery 2008 86, 1268-1276DOI: (10.1016/j.athoracsur.2008.04.107) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Zymographic bands for matrix metalloproteinase (MMP)-9 and MMP-2 were detected in all myocardial samples for the myocardial infarct (MI), border, and remote regions at 92 kDa (MMP-9) and at 64 to 72 kDa, consistent with that for the latent and active forms of MMP-2. (Top) MMP-9 levels were increased from reference control values in the border and MI regions of both MI groups. (Bottom) Within the MI region, levels of activated MMP-2 were lower in the fibrin-alginate (Fib-Alg) group than the saline group. (C = p < 0.05 vs reference controls; R = p < 0.05 vs treatment-matched remote region; B = p < 0.05 vs treatment-matched border region; *p < 0.05 vs same region of the saline group). Data are presented as the mean and standard error of the mean. The Annals of Thoracic Surgery 2008 86, 1268-1276DOI: (10.1016/j.athoracsur.2008.04.107) Copyright © 2008 The Society of Thoracic Surgeons Terms and Conditions