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Functional and Biochemical Assessment of Porcine Hearts After Simulated Donation After Circulatory Death
S150
The Journal of Heart and Lung Transplantation, Vol 34, No 4S, April 2015
Results: Bacterial count significantly decreased by 12h of EVLP in the ABx group, compared to controls. TNF-α , IL-1β , MIP-1α and MIP-1β at 12h were significantly lower in the cases where all the bacteria decreased (P ≤ 0.05). Endotoxin (> 2 EU/mL) was detected in 5/8 cases in the ABx group and 3/7 cases in the control group. Endotoxin level at 12h was significantly lower in the ABx group, compared to the control group (3.83±1.54 vs. 397±397 EU/mL; P = 0.0357). Importantly, TNF-α , IL-1β , IL-6, IFN-γ , IL-4, IL-5, IL-9, IL-10, IL-8, G-CSF, MCP-1, MIP-1α and VEGF at 12h were significantly higher in endotoxin positive cases, compared with negative cases (Figure 1; P ≤ 0.05). TNF-α , IL-1β , MIP-1α , MIP-1β , IL-9 and VEGF levels at 12h were strongly correlated with endotoxin levels at 12h (Spearman R > 0.70). Conclusion: BAL bacterial count response to antibiotic therapy and EVLP perfusate levels of key inflammatory mediators (TNF-α , IL-1β , MIP-1α ) were significantly correlated to observed perfusate endotoxin levels.
3( 90) 3( 89) Functional and Biochemical Assessment of Porcine Hearts After Simulated Donation After Circulatory Death M.A. Schechter , K.W. Southerland, B.J. Feger, L. Njoroge, D. Linder Jr., D.E. Bowles, C.A. Milano. Surgery, Duke University Medical Center, Durham, NC. Purpose: Donation after circulatory death (DCD) has the potential to increase the number of hearts available for transplantation. However, the added warm ischemic injury with DCD allografts could be limiting. The purpose of this study was to examine the biochemical and functional impact of DCD using a porcine model. Methods: 70 Kg Yorkshire pigs were used for all experiments, and baseline in vivo cardiac function was established prior to heart procurement. In the DCD group (n = 5), death was induced by cessation of mechanical ventilation, and the hearts were procured after a 10 minute stand-off period following circulatory arrest. In the control group (n = 5), the hearts were procured in a from a beating donor in the conventional fashion. All hearts were flushed with ice cold preservation solution (Celsoir) and placed in cold static storage for two hours. After storage, hearts were perfused on an ex-vivo circuit, which was then placed into working mode. Graft function was assessed with and without dobutamine (5 mcg/kg/min). The concentration of cardiac troponin I while on the ex-vivo circuit was measured and normalized to heart weight. Results: In vivo cardiac function was identical for both groups. The DCD hearts exhibited significantly more loss of function after preservation and were less responsive to high-dose inotropes compared to the control group (Figure 1a). These findings correlated with the biochemical assessment, which showed that while the two groups are fairly similar during the early reperfusion period, once the hearts are put into working mode, the DCD heart have a much larger rise in troponins (Figure 1b). Conclusion: In a porcine model, DCD is associated with increased myocardial damage compared to standard, beating heart donors. However, DCD hearts were able to perform work. This finding suggests that DCD hearts could be a potential source of organs for donation in the future, but may require strategies to mitigate ischemic injury.
Three-Dimensional Human-Heart Derived Scaffolds A.N. Patel , J. Theisen, A.A. Winters, D. Grainger, F. Silva, D. Bull. University of Utah, Salt Lake City, UT. Purpose: Heart failure results from damaged myocardium that can severely reduce function and lead to death with very few options for treatment. Recent studies have injected stem cells into the coronary arteries to improve function. Though the potential of stem cell therapy is great, only limited efficacy has been attained, Extracellular matrix (ECM) proteins derived ECM from bowel or bladder and stem cells in concert may have superior benefits to either alone but the homing cues are still limited for engraftment due to lack of organ specific signaling and xenogenic issues. Our goal was to derive a novel three-dimensional human heart extracellular matrix (ECM)-derived scaffold that could serve as a vehicle to deliver cardiac or stem cells directly to the damaged tissue of the heart and remain at the treatment site. Methods: Scaffolds were created from purified human heart ECM proteins to expedite clinical translation and actively promote and preserve heart cell phenotype. Scaffolds were imparted with pores 250 um in size to enhance diffusion and cell penetration and distribution. Human cardiomyocytes and cardiac derived-iPSCs were seeded into scaffolds 3 mm in diameter and 0.3 mm thick, cultured and subsequently implanted into NOD-SCID mice. Results: The human cardiomyocytes and cardiac derived iPSCs readily adhered to human cardiac-derived ECM protein scaffolds. Cardiomoycytes within the scaffold maintained representative phenotypes including expression of cardiomyocyte-specific markers and remained electrically active, even beating the scaffold in unison in vitro. In vivo, cardiomyocyte-seeded scaffolds spontaneously adhered to and incorporated with murine infarcted tissue. Conclusion: These results indicate that a novel 3D scaffold made of purified human cardiac ECM can be used as a delivery vehicle for human cardiac cells to directly target damaged heart tissue. Optimization of scaffolds and cell interactions need to be studied further in vivo. 3( 91) Porcine B4GALNT2 a Source of New Xenogenic Glycan G.W. Byrne ,1 Z. Du,2 H. Kogelberg,1 C. McGregor.1 1Cardiovascular Science, University College London, London, United Kingdom; 2Department of Surgery, Mayo Clinic, Rochester, MN.
The Journal of Heart and Lung Transplantation, Vol 34, No 4S, April 2015
Results: Bacterial count significantly decreased by 12h of EVLP in the ABx group, compared to controls. TNF-α , IL-1β , MIP-1α and MIP-1β at 12h were significantly lower in the cases where all the bacteria decreased (P ≤ 0.05). Endotoxin (> 2 EU/mL) was detected in 5/8 cases in the ABx group and 3/7 cases in the control group. Endotoxin level at 12h was significantly lower in the ABx group, compared to the control group (3.83±1.54 vs. 397±397 EU/mL; P = 0.0357). Importantly, TNF-α , IL-1β , IL-6, IFN-γ , IL-4, IL-5, IL-9, IL-10, IL-8, G-CSF, MCP-1, MIP-1α and VEGF at 12h were significantly higher in endotoxin positive cases, compared with negative cases (Figure 1; P ≤ 0.05). TNF-α , IL-1β , MIP-1α , MIP-1β , IL-9 and VEGF levels at 12h were strongly correlated with endotoxin levels at 12h (Spearman R > 0.70). Conclusion: BAL bacterial count response to antibiotic therapy and EVLP perfusate levels of key inflammatory mediators (TNF-α , IL-1β , MIP-1α ) were significantly correlated to observed perfusate endotoxin levels.
3( 90) 3( 89) Functional and Biochemical Assessment of Porcine Hearts After Simulated Donation After Circulatory Death M.A. Schechter , K.W. Southerland, B.J. Feger, L. Njoroge, D. Linder Jr., D.E. Bowles, C.A. Milano. Surgery, Duke University Medical Center, Durham, NC. Purpose: Donation after circulatory death (DCD) has the potential to increase the number of hearts available for transplantation. However, the added warm ischemic injury with DCD allografts could be limiting. The purpose of this study was to examine the biochemical and functional impact of DCD using a porcine model. Methods: 70 Kg Yorkshire pigs were used for all experiments, and baseline in vivo cardiac function was established prior to heart procurement. In the DCD group (n = 5), death was induced by cessation of mechanical ventilation, and the hearts were procured after a 10 minute stand-off period following circulatory arrest. In the control group (n = 5), the hearts were procured in a from a beating donor in the conventional fashion. All hearts were flushed with ice cold preservation solution (Celsoir) and placed in cold static storage for two hours. After storage, hearts were perfused on an ex-vivo circuit, which was then placed into working mode. Graft function was assessed with and without dobutamine (5 mcg/kg/min). The concentration of cardiac troponin I while on the ex-vivo circuit was measured and normalized to heart weight. Results: In vivo cardiac function was identical for both groups. The DCD hearts exhibited significantly more loss of function after preservation and were less responsive to high-dose inotropes compared to the control group (Figure 1a). These findings correlated with the biochemical assessment, which showed that while the two groups are fairly similar during the early reperfusion period, once the hearts are put into working mode, the DCD heart have a much larger rise in troponins (Figure 1b). Conclusion: In a porcine model, DCD is associated with increased myocardial damage compared to standard, beating heart donors. However, DCD hearts were able to perform work. This finding suggests that DCD hearts could be a potential source of organs for donation in the future, but may require strategies to mitigate ischemic injury.
Three-Dimensional Human-Heart Derived Scaffolds A.N. Patel , J. Theisen, A.A. Winters, D. Grainger, F. Silva, D. Bull. University of Utah, Salt Lake City, UT. Purpose: Heart failure results from damaged myocardium that can severely reduce function and lead to death with very few options for treatment. Recent studies have injected stem cells into the coronary arteries to improve function. Though the potential of stem cell therapy is great, only limited efficacy has been attained, Extracellular matrix (ECM) proteins derived ECM from bowel or bladder and stem cells in concert may have superior benefits to either alone but the homing cues are still limited for engraftment due to lack of organ specific signaling and xenogenic issues. Our goal was to derive a novel three-dimensional human heart extracellular matrix (ECM)-derived scaffold that could serve as a vehicle to deliver cardiac or stem cells directly to the damaged tissue of the heart and remain at the treatment site. Methods: Scaffolds were created from purified human heart ECM proteins to expedite clinical translation and actively promote and preserve heart cell phenotype. Scaffolds were imparted with pores 250 um in size to enhance diffusion and cell penetration and distribution. Human cardiomyocytes and cardiac derived-iPSCs were seeded into scaffolds 3 mm in diameter and 0.3 mm thick, cultured and subsequently implanted into NOD-SCID mice. Results: The human cardiomyocytes and cardiac derived iPSCs readily adhered to human cardiac-derived ECM protein scaffolds. Cardiomoycytes within the scaffold maintained representative phenotypes including expression of cardiomyocyte-specific markers and remained electrically active, even beating the scaffold in unison in vitro. In vivo, cardiomyocyte-seeded scaffolds spontaneously adhered to and incorporated with murine infarcted tissue. Conclusion: These results indicate that a novel 3D scaffold made of purified human cardiac ECM can be used as a delivery vehicle for human cardiac cells to directly target damaged heart tissue. Optimization of scaffolds and cell interactions need to be studied further in vivo. 3( 91) Porcine B4GALNT2 a Source of New Xenogenic Glycan G.W. Byrne ,1 Z. Du,2 H. Kogelberg,1 C. McGregor.1 1Cardiovascular Science, University College London, London, United Kingdom; 2Department of Surgery, Mayo Clinic, Rochester, MN.