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Ice-free cryopreservation of heart valve allografts: better extracellular matrix preservation in vivo and preclinical results |
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| Authors: | Kelvin G. M. Brockbank Katja Schenke-Layland Elizabeth D. Greene Zhenzhen Chen Olaf Fritze Martina Schleicher Renate Kaulitz Iris Riemann Falko Fend Johannes M. Albes Ulrich A. Stock Milan Lisy |
| Affiliation: | 1. Cell and Tissue Systems, Inc., 2231 Technical Parkway, Suite A, North Charleston, SC, 29406, USA 2. Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USA 3. Deparment of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA 4. Cardiovascular Research Laboratories, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA 5. Deparment of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Stuttgart, Germany 6. Inter-University Centre for Medical Technology Stuttgart-Tübingen (IZST), Eberhard Karls University Tübingen, Tübingen, Germany 7. Deparment of Thoracic, Cardiac and Vascular Surgery, University Hospital Tuebingen, Tuebingen, Germany 8. Deparment of Pediatric Cardiology, University Hospital Tuebingen, Tuebingen, Germany 9. Fraunhofer Institute of Biomedical Technology, St. Ingbert, Germany 10. Deparment of General Pathologie and Pathological Anatomy, University Hospital Tuebingen, Tuebingen, Germany 11. Deparment of Cardiac Surgery, Heart Centre Brandenburg, Bernau, Germany |
| Abstract: | The purpose of this study was evaluation of an ice-free cryopreservation method for heart valves in an allogeneic juvenile pulmonary sheep implant model and comparison with traditionally frozen cryopreserved valves. Hearts of 15 crossbred Whiteface sheep were procured in Minnesota. The valves were processed in South Carolina and the pulmonary valves implanted orthotopically in 12 black faced Heidschnucke sheep in Germany. The ice-free cryopreserved valves were cryopreserved in 12.6?mol/l cryoprotectant (4.65, 4.65, and 3.31?mol/l of dimethylsulfoxide, formamide and 1,2-propanediol) and stored at ?80°C. Frozen valves were cryopreserved by controlled slow rate freezing in 1.4?mol/l dimethylsulfoxide and stored in vapor-phase nitrogen. Aortic valve tissues were used to evaluate the impact of preservation without implantation. Multiphoton microscopy revealed reduced but not significantly damaged extracellular matrix before implantation in frozen valves compared with ice-free tissues. Viability assessment revealed significantly less metabolic activity in the ice-free valve leaflets and artery samples compared with frozen tissues (P?P?=?0.0403, P?=?0.0591). Histo-pathology showed significantly thickened leaflets in the frozen valves (P?P? |
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