Anja Lena Thiebes1, Manuel Armin Reddemann2, Johannes Palmer2, Reinhold Kneer2, Stefan Jockenhoevel1,3, Christian Gabriel Cornelissen1,4. 1. 1 Department of Tissue Engineering and Textile Implants, Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University , Aachen, Germany . 2. 2 Institute of Heat and Mass Transfer, RWTH Aachen University , Aachen, Germany . 3. 3 Aachen-Maastricht Institute for Biobased Materials, Maastricht University at Chemelot Campus , Geleen, The Netherlands . 4. 4 Section for Pneumology, Department for Internal Medicine, RWTH Aachen University , Aachen, Germany .
Abstract
INTRODUCTION: Inoperable airway stenoses are currently treated by placing stents. A major problem of covered stents is missing mucociliary clearance, which is caused by covering the native respiratory epithelium. By coating a stent with respiratory epithelium, this problem can be overcome. However, no methods are available for efficient endoscopic cell seeding. METHODS: We designed a flexible endoscopic spraying device based on a bronchoscope and tested it with respiratory epithelial cells. With this device cells can also be applied in a thin layer of fibrin glue. We evaluated the survival rate directly after spray application with a live-dead staining and the long-term differentiation capacity with histology and electron microscopy. Furthermore, the random distribution of cells when applied in a tube was analyzed and the macroscopic and microscopic characteristics of the endoscopic spray were investigated using high-speed visualization. RESULTS: Spray visualization revealed a polydisperse character of the spray with the majority of droplets larger than epithelial cells. Spray application does not influence the survival rate and differentiation of respiratory epithelial cells. After 4 weeks, cells built up a pseudostratified epithelial layer with cilia and goblet cells. When cells are applied in a thin layer of fibrin gel into a tube, a nearest neighbor index of 1.2 is obtained, which suggests a random distribution of the cells. CONCLUSIONS: This spraying device is a promising tool for application of various cell types onto stents or implants with high survival rates and homogeneous distribution as shown in this study for ovine respiratory epithelial cells. The system could also be used for cell therapy to locally apply cells to the diseased parts of hollow organs. For the first time, the fluid dynamics of a spray device for cells were examined to validate in vitro results.
INTRODUCTION: Inoperable airway stenoses are currently treated by placing stents. A major problem of covered stents is missing mucociliary clearance, which is caused by covering the native respiratory epithelium. By coating a stent with respiratory epithelium, this problem can be overcome. However, no methods are available for efficient endoscopic cell seeding. METHODS: We designed a flexible endoscopic spraying device based on a bronchoscope and tested it with respiratory epithelial cells. With this device cells can also be applied in a thin layer of fibrin glue. We evaluated the survival rate directly after spray application with a live-dead staining and the long-term differentiation capacity with histology and electron microscopy. Furthermore, the random distribution of cells when applied in a tube was analyzed and the macroscopic and microscopic characteristics of the endoscopic spray were investigated using high-speed visualization. RESULTS: Spray visualization revealed a polydisperse character of the spray with the majority of droplets larger than epithelial cells. Spray application does not influence the survival rate and differentiation of respiratory epithelial cells. After 4 weeks, cells built up a pseudostratified epithelial layer with cilia and goblet cells. When cells are applied in a thin layer of fibrin gel into a tube, a nearest neighbor index of 1.2 is obtained, which suggests a random distribution of the cells. CONCLUSIONS: This spraying device is a promising tool for application of various cell types onto stents or implants with high survival rates and homogeneous distribution as shown in this study for ovine respiratory epithelial cells. The system could also be used for cell therapy to locally apply cells to the diseased parts of hollow organs. For the first time, the fluid dynamics of a spray device for cells were examined to validate in vitro results.
Authors: Stefan Weinandy; Lisanne Rongen; Fabian Schreiber; Christian Cornelissen; Thomas Cormac Flanagan; Andreas Mahnken; Thomas Gries; Thomas Schmitz-Rode; Stefan Jockenhoevel Journal: Tissue Eng Part A Date: 2012-06-07 Impact factor: 3.845
Authors: Herve Dutau; Ali I Musani; Jerome Plojoux; Sophie Laroumagne; Philippe Astoul Journal: Expert Rev Respir Med Date: 2014-01-22 Impact factor: 3.772
Authors: Alexander Kaminski; Christian Klopsch; Peter Mark; Can Yerebakan; Peter Donndorf; Ralf Gäbel; Friederike Eisert; Stefan Hasken; Sebastian Kreitz; Aenne Glass; Stefan Jockenhövel; Nan Ma; Guenther Kundt; Andreas Liebold; Gustav Steinhoff Journal: Tissue Eng Part C Methods Date: 2010-11-22 Impact factor: 3.056
Authors: Ludovic Zimmerlin; J Peter Rubin; Melanie E Pfeifer; Linda R Moore; Vera S Donnenberg; Albert D Donnenberg Journal: Cytotherapy Date: 2013-01 Impact factor: 5.414
Authors: Anja Lena Thiebes; Stefanie Albers; Christian Klopsch; Stefan Jockenhoevel; Christian G Cornelissen Journal: Biores Open Access Date: 2015-06-01
Authors: Dana M Schwartz; Meryem O Pehlivaner Kara; Allan M Goldstein; Harald C Ott; Adam K Ekenseair Journal: Tissue Eng Part C Methods Date: 2017-09 Impact factor: 3.056