Michael P Francis1, Erick Breathwaite2, Anna A Bulysheva3, Frency Varghese4, Rudy U Rodriguez5, Sucharita Dutta6, Iurii Semenov7, Rebecca Ogle8, Alexander Huber9, Alexandra-Madelaine Tichy10, Silvia Chen11, Christian Zemlin12. 1. LifeNet Health Institute of Regenerative Medicine, Virginia Beach, VA, United States; Eastern Virginia Medical School, Norfolk, VA, United States. Electronic address: mpf3b@virginia.edu. 2. LifeNet Health Institute of Regenerative Medicine, Virginia Beach, VA, United States. Electronic address: erick_breathwaite@lifenethealth.org. 3. Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, United States. Electronic address: abulyshe@odu.edu. 4. Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, United States; Department of Electrical and Computer Engineering, Old Dominion University, Norfolk, VA, United States. Electronic address: fvarg001@odu.edu. 5. LifeNet Health Institute of Regenerative Medicine, Virginia Beach, VA, United States. Electronic address: rudy_rodriguez@lifenethealth.org. 6. Eastern Virginia Medical School, Norfolk, VA, United States. Electronic address: duttasm@evms.edu. 7. Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, United States. 8. LifeNet Health Institute of Regenerative Medicine, Virginia Beach, VA, United States. Electronic address: Rebecca_ogle@lifenethealth.org. 9. LifeNet Health Institute of Regenerative Medicine, Virginia Beach, VA, United States. Electronic address: alexander_huber@lifenethealth.org. 10. FH Campus Wien, Applied Life Sciences, Wien, Austria. 11. LifeNet Health Institute of Regenerative Medicine, Virginia Beach, VA, United States. 12. Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, United States; Department of Electrical and Computer Engineering, Old Dominion University, Norfolk, VA, United States. Electronic address: czemlin@odu.edu.
Abstract
INTRODUCTION: Xenogeneic extracellular matrix (ECM) hydrogels have shown promise in remediating cardiac ischemia damage in animal models, yet analogous human ECM hydrogels have not been well development. An original human placenta-derived hydrogel (hpECM) preparation was thus generated for assessment in cardiomyocyte cell culture and therapeutic cardiac injection applications. METHODS AND RESULTS: Hybrid orbitrap-quadrupole mass spectrometry and ELISAs showed hpECM to be rich in collagens, basement membrane proteins, and regenerative growth factors (e.g. VEGF-B, HGF). Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes synchronized and electrically coupled on hpECM faster than on conventional cell culture environments, as validated by intracellular calcium measurements. In vivo, injections using biotin-labeled hpECM confirmed its spatially discrete localization to the myocardium proximal to the injection site. hpECM was injected into rat myocardium following an acute myocardium infarction induced by left anterior descending artery ligation. Compared to sham treated animals, which exhibited aberrant electrical activity and larger myocardial scars, hpECM injected rat hearts showed a significant reduction in scar volume along with normal electrical activity of the surviving tissue, as determined by optical mapping. CONCLUSION: Placental matrix and growth factors can be extracted as a hydrogel that effectively supports cardiomyocytes in vitro, and in vivo reduces scar formation while maintaining electrophysiological activity when injected into ischemic myocardium. STATEMENT OF SIGNIFICANCE: This is the first report of an original extracellular matrix hydrogel preparation isolated from human placentas (hpECM). hpECM is rich in collagens, laminin, fibronectin, glycoproteins, and growth factors, including known pro-regenerative, pro-angiogenic, anti-scarring, anti-inflammatory, and stem cell-recruiting factors. hpECM supports the culture of cardiomyocytes, stem cells and blood vessels assembly from endothelial cells. In a rat model of myocardial infarction, hpECM injections were safely deliverable to the ischemic myocardium. hpECM injections repaired the myocardium, resulting in a significant reduction in infarct size, more viable myocardium, and a normal electrophysiological contraction profile. hpECM thus has potential in therapeutic cardiovascular applications, in cellular therapies (as a delivery vehicle), and is a promising biomaterial for advancing basic cell-based research and regenerative medicine applications.
INTRODUCTION: Xenogeneic extracellular matrix (ECM) hydrogels have shown promise in remediating cardiac ischemia damage in animal models, yet analogous human ECM hydrogels have not been well development. An original human placenta-derived hydrogel (hpECM) preparation was thus generated for assessment in cardiomyocyte cell culture and therapeutic cardiac injection applications. METHODS AND RESULTS: Hybrid orbitrap-quadrupole mass spectrometry and ELISAs showed hpECM to be rich in collagens, basement membrane proteins, and regenerative growth factors (e.g. VEGF-B, HGF). Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes synchronized and electrically coupled on hpECM faster than on conventional cell culture environments, as validated by intracellular calcium measurements. In vivo, injections using biotin-labeled hpECM confirmed its spatially discrete localization to the myocardium proximal to the injection site. hpECM was injected into rat myocardium following an acute myocardium infarction induced by left anterior descending artery ligation. Compared to sham treated animals, which exhibited aberrant electrical activity and larger myocardial scars, hpECM injected rat hearts showed a significant reduction in scar volume along with normal electrical activity of the surviving tissue, as determined by optical mapping. CONCLUSION: Placental matrix and growth factors can be extracted as a hydrogel that effectively supports cardiomyocytes in vitro, and in vivo reduces scar formation while maintaining electrophysiological activity when injected into ischemic myocardium. STATEMENT OF SIGNIFICANCE: This is the first report of an original extracellular matrix hydrogel preparation isolated from human placentas (hpECM). hpECM is rich in collagens, laminin, fibronectin, glycoproteins, and growth factors, including known pro-regenerative, pro-angiogenic, anti-scarring, anti-inflammatory, and stem cell-recruiting factors. hpECM supports the culture of cardiomyocytes, stem cells and blood vessels assembly from endothelial cells. In a rat model of myocardial infarction, hpECM injections were safely deliverable to the ischemic myocardium. hpECM injections repaired the myocardium, resulting in a significant reduction in infarct size, more viable myocardium, and a normal electrophysiological contraction profile. hpECM thus has potential in therapeutic cardiovascular applications, in cellular therapies (as a delivery vehicle), and is a promising biomaterial for advancing basic cell-based research and regenerative medicine applications.
Authors: Peter A Mollica; Elizabeth N Booth-Creech; John A Reid; Martina Zamponi; Shea M Sullivan; Xavier-Lewis Palmer; Patrick C Sachs; Robert D Bruno Journal: Acta Biomater Date: 2019-06-21 Impact factor: 8.947
Authors: Paula Fratini; Nathia Nathaly Rigoglio; Gustavo de Sá Schiavo Matias; Ana Claudia O Carreira; Rose Eli Grassi Rici; Maria Angelica Miglino Journal: Biores Open Access Date: 2018-07-01