List of publications Kevin Tröndle

• Journal Articles
• Conference papers

Journal Articles

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2023

• J. Weygant, F. Koch, K. Adam, K. Tröndle, R. Zengerle, G. Finkenzeller, S. Kartmann, P. Koltay, S. Zimmermann
  A Drop-on-Demand Bioprinting Approach to Spatial Arrangement of Multiple Cell Types and Monitoring Their Cell–Cell Interactions towards Vascularization Based on Endothelial Cells and Mesenchymal Stem Cells
  2023 Cells, volume: 12, issue: 4, page: 646

2022

• K. Tröndle, G. Miotto, L. Rizzo, R. Pichler, F. Koch, P. Koltay, R. Zengerle, S. S. Lienkamp, S. Kartmann, S. Zimmermann
  Deep Learning-Assisted Nephrotoxicity Testing with Bioprinted Renal Spheroids
  2022 Int. J. Bioprint, volume: 8, issue: 2, page: 528
• F. Koch, O. Thaden, S. Conrad, K. Tröndle, G. Finkenzeller, R. Zengerle, S. Kartmann, S. Zimmermann, P. Koltay
  Mechanical properties of polycaprolactone (PCL) scaffolds for hybrid 3D-bioprinting with alginate-gelatin hydrogel
  2022 J. Mech. Behav. Biomed. Mat., volume: 130, page: 105219

2021

• F. Koch, O. Thaden, K. Tröndle, R. Zengerle, S. Zimmermann, P. Koltay
  Open-source hybrid 3D-bioprinter for simultaneous printing of thermoplastics and hydrogels
  2021 HardwareX, volume: 10, page: e00230

2020

• F. Koch, K. Tröndle, G. Finkenzeller, R. Zengerle, S. Zimmermann, P. Koltay
  Generic method of printing window adjustment for extrusion-based 3D-bioprinting to maintain high viability of mesenchymal stem cells in an alginate-gelatin hydrogel
  2020 Bioprinting, page: e00094
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  Abstract

  Over the last decade, bioprinting of artificial tissues has been developed into a significant field of research. With an increasing number of printing technologies and bioinks used in bioprinting, its complexity increases as both the printing technology and the properties of the bioink influence the cell biological functionality and printing accuracy of the printed tissue. Therefore, optimization of bioprinting processes often remains a challenge, which could be solved by a smart fine-tuning of the process parameters. We present a novel method to adjust the printing window for extrusion-based bioprinting on the basis of a two-step assessment to determine process parameters such as nozzle size, extrusion flow rate, and printing temperature. First, a suitable printing temperature is deduced from the bioink properties and second nozzle size and extrusion flow rate is selected in a way that the immediate cell damage after printing is reduced. For both steps only basic rheological properties of the bioinks need to be known as well as detailed knowledge of the cell survival in the bioink for different shear stresses. This method is applied to an exemplary alginate-gelatin hydrogel to show how the printing temperature affects the achievable printing accuracy. For this bioink, viability of immortalized mesenchymal stem cells (iMSC) decreases with about 4% per thousand Pascal increase in maximum shear stress. For different combinations of flow rate, nozzle size and nozzle shape it is shown, that only the maximum shear stress experienced by the iMSCs influences average cell viability. Factors like flow rate, nozzle size and shape only play an indirect role by influencing the maximum shear stress and individually have no significant influence on cell viability. The experimental results allow a direct adjustment of printing parameters for the presented combination of hydrogel and cell type but are not limited to it. For other bioinks, the described generic method can be easily used to systematically adjust the printing parameters. For this purpose, only the basic rheological properties and the influence of shear stress on cell survival need to be known and process parameters can be set concerning the respective application.
• P. Rukavina, F. Koch, M. Wehrle, K. Tröndle, G. B. Stark, P. Koltay, S. Zimmermann, R. Zengerle, F. Lampert, S. Strassburg, G. Finkenzeller, F. Simunovic
  In vivo evaluation of bioprinted prevascularized bone tissue
  2020 Biotechnol Bioeng, pages: 1 - 10
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  Abstract

  Bioprinting can be considered as a progression of the classical tissue engineering approach, in which cells are randomly seeded into scaffolds. Bioprinting offers the advantage that cells can be placed with high spatial fidelity within three‐dimensional tissue constructs. A decisive factor to be addressed for bioprinting approaches of artificial tissues is that almost all tissues of the human body depend on a functioning vascular system for the supply of oxygen and nutrients. In this study, we have generated cuboid prevascularized bone tissue constructs by bioprinting human adipose‐derived mesenchymal stem cells (ASCs) and human umbilical vein endothelial cells (HUVECs) by extrusion‐based bioprinting and drop‐on‐demand (DoD) bioprinting, respectively. The computer‐generated print design could be verified in vitro after printing. After subcutaneous implantation of bioprinted constructs in immunodeficient mice, blood vessel formation with human microvessels of different calibers could be detected arising from bioprinted HUVECs and stabilization of human blood vessels by mouse pericytes was observed. In addition, bioprinted ASCs were able to synthesize a calcified bone matrix as an indicator of ectopic bone formation. These results indicate that the combined bioprinting of ASCs and HUVECs represents a promising strategy to produce prevascularized artificial bone tissue for prospective applications in the treatment of critical‐sized bone defects.

2019

• K. Tröndle, F. Koch, G. Finkenzeller, G. B. Stark, R. Zengerle, P. Koltay, S. Zimmermann
  Bioprinting of high cell density constructs leads to controlled lumen formation with self‐assembly of endothelial cells
  2019 Journal of Tissue Engineering and Regenerative Medicine, volume: 13, issue: 10, pages: 1883 - 1895
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  Abstract

  Active nutrient supply and waste product removal are key requirements for the fabrication of long term viable and functional tissue constructs of considerable size. This work aims to contribute to the fabrication of artificial perfusable networks with a bioprinting process, based on drop‐on‐demand (DoD) printing of primary endothelial cell (EC) suspension bioink (25 · 106 ± 3 · 106 cells/ml). The process results in prescribed lumen between two hydrogel layers, allowing its integration in common layering based bioprinting processes. Low volume bioink droplets (appr. 10 nl) as building blocks, were deposited between two fibrin or collagen I layers to realize shapeable, cell‐rich aggregates. Unattainable with manual positioning, DoD printing allowed precise fabrication of various designs, such as spheroidal‐, line‐shaped and Y‐branch cellular structures, with a mean lateral extension of 285 ± 81 μm. For basic characterization, the cell suspension building blocks were systematically compared to preformed spheroids of the same cell type, passage and number. Post printing investigations of initially loose cell arrangements showed self‐assembly and formation of central lumen with a mean cross‐sectional area of Ølumen = 6400 μm2 at day 3, lined by a single layer of CD31 positive ECs, as evaluated by confocal microscopy. Originating from this main lumen smaller, undirected side‐branches (Øbranches = 740 μm2) were formed by sprouting cells, inducing a first step towards a simplistic hierarchically organized network. These lumen could prospectively help for tissue construct perfusion in vitro or, potentially, as niche for angiogenesis of host vascularization in implants.

2017

• L. Benning, L. Gutzweiler, K. Tröndle, J. Riba, R. Zengerle, P. Koltay, S. Zimmermann, G. B. Stark, G. Finkenzeller
  Assessment of hydrogels for bioprinting of endothelial cells
  2017 J Biomed Mater Res A, pages: 935 - 947
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  Abstract

  In tissue engineering applications, vascularization can be accomplished by co-implantation of tissue forming cells and endothelial cells (ECs), whereby the latter are able to form functional blood vessels. The use of three-dimensional (3D) bioprinting technologies has the potential to improve the classical tissue engineering approach because these will allow the generation of scaffolds with high spatial control of endothelial cell allocation. This study focuses on a side by side comparisons of popular commercially available bioprinting hydrogels (matrigel, fibrin, collagen, gelatin, agarose, Pluronic F-127, alginate and alginate/gelatin) in the context of their physicochemical parameters, their swelling/degradation characteristics, their biological effects on vasculogenesis-related EC parameters and their printability. The aim of this study was to identify the most suitable hydrogel or hydrogel combination for inkjet printing of ECs to build pre-vascularized tissue constructs. Most tested hydrogels displayed physicochemical characteristics suitable for inkjet printing. However, Pluronic F-127 and the alginate/gelatin blend were rapidly degraded when incubated in cell culture medium. Agarose, Pluronic F-127, alginate and alginate/gelatin hydrogels turned out to be unsuitable for bioprinting of ECs because of their non-adherent properties and/or their incapability to support EC proliferation. Gelatin was able to support EC proliferation and viability but was unable to support endothelial cell sprouting. Our experiments revealed fibrin and collagen to be most suitable for bioprinting of ECs, because these hydrogels showed acceptable swelling/degradation characteristics, supported vasculogenesis-related EC parameters and showed good printability. Moreover, ECs in constructs of preformed spheroids survived the printing process and formed capillary-like cords.
• L. Benning, L. Gutzweiler, K. Tröndle, J. Riba, R. Zengerle, P. Koltay, S. Zimmermann, G.B. Stark, G. Finkenzeller
  Cytocompatibility testing of hydrogels toward bioprinting of mesenchymal stem cells
  2017 J Biomed Mater Res A, volume: 105, pages: 3231 - 3241
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  Abstract

  Mesenchymal stem cells (MSCs) represent a very attractive cell source for tissue engineering applications aiming at the generation of artificial bone substitutes. The use of three-dimensional bioprinting technologies has the potential to improve the classical tissue engineering approach because bioprinting will allow the generation of hydrogel scaffolds with high spatial control of MSC allocation within the bioprinted construct. In this study, we have performed direct comparisons between commercially available hydrogels in the context of their cytocompatibility toward MSCs and their physicochemical parameters with the aim to identify the most suitable hydrogel for drop-on-demand (DoD) printing of MSCs. In this context, we examined matrigel, fibrin, collagen, gelatin, and gelatin/alginate at various hydrogel concentrations. Matrigel, fibrin, collagen, and gelatin were able to support cell viability, but the latter showed a limited potential to promote MSC proliferation. We concentrated our study on fibrin and collagen hydrogels and investigated the effect of hydroxyapatite (HA) inclusion. The inclusion of HA enhanced proliferation and osteogenic differentiation of MSCs and prevented degradation of fibrin in vitro. According to viscosity and storage moduli measurements, HA-blends displayed physicochemical characteristics suitable for DoD printing. In bioprinting experiments, we confirmed that fibrin and collagen and their respective HA-blends represent excellent hydrogels for DoD-based printing as evidenced by high survival rates of printed MSCs.
• L. Gutzweiler, S. Kartmann, K. Troendle, L. Benning, G. Finkenzeller, R. Zengerle, P. Koltay, B. Stark, S. Zimmermann
  Large scale production and controlled deposition of single HUVEC spheroids for bioprinting applications
  2017 Biofabrication, volume: 9 (2), page: 02502
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  Abstract

  We present 1.) a fast and automated method for large scale production of HUVEC spheroids based on the hanging drop method and 2.) a novel method for well-controlled lateral deposition of single spheroids by drop-on-demand printing. Large scale spheroid production is achieved via printing 1536 droplets of HUVEC cell suspension having a volume of 1 µl each within 3 minutes at a pitch of 2.3 mm within an array of 48 x 32 droplets onto a flat substrate. Printing efficiencies between 97.9% and 100% and plating efficiencies between 87.3% and 100% were achieved. Harvested spheroids (consisting of approx. 250 HUVECs each) appear uniform in size and shape. After incubation and harvesting, the spheroids are deposited individually in user-defined patterns onto hydrogels using an automated drop-on-demand dispenser setup. Controlled by an image detection algorithm focusing the dispenser nozzle, droplets containing exactly one spheroid are printed onto a substrate, while all other droplets are discarded. Using this approach an array of 6 x 3 HUVEC spheroids with intermediate distances of 500 µm embedded in fibrin was generated. Successful progress of spheroid sprouting and merging of neighboring sprouts was observed during the first 72 hours of incubation indicating a good viability of the deposited spheroids.

Conference papers

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2021

• J. Weygant, F. Koch, K. Troendle, G. Finkenzeller, R. Zengerle, S. Kartmann, S. Zimmermann, P. Koltay
  Drop-on-Demand Bioprinting Approach For Precise Alignment and Interaction Studies of Different Cell Types
  2021 International Conference on Biofabrication, Australia, 27. – 29.09.2021 (online)
• K. Tröndle, A. Itani, F. Koch, R. Zengerle, P. Koltay, S. Zimmermann
  Drop-on-demand bioprinting solutions for the fabrication of 3D cell culture systems
  2021 DECHEMA 3D Cell Culture Freiburg (online), 05.-07.05.2021
• F. Koch, K. Tröndle, G. Finkenzeller, P. Rukavina, R. Zengerle, P. Koltay, S. Zimmermann
  Hybrider 3D-Biodruck zur künstlichen Herstellung von Knochen / Using hybrid processes for 3D-bioprinting of artificial bone tissue
  2021 MST-Kongress, Ludwigsburg, 08.-10.11.2021

2020

• K. Tröndle, A. Itani, F. Koch, R. Zengerle, S. Zimmermann, P. Koltay
  Fabrication and fluidic integration of self-assembled cellular microtubules for nephron-on-chip applications
  2020 MicroTAS 2020, 04.-09.10.2020, virtual

2019

• F. Koch, M. Wehrle, K. Tröndle, P. Koltay, G. Finkenzeller, R. Zengerle, S. Zimmermann
  Rapid assessment of combined drop on demand and extrusion-based bioprinting with controlled shear stress and high shape fidelity
  2019 Transducers 2019 - EUROSENSORS XXXIII 23. -27. Juni 2019 - Berlin, Germany
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  Abstract

  We present a novel combination of drop on demand (DoD) and extrusion-based bioprinting to generate highprecision patterns of cells inside large hydrogel volumes. Extrusion-based bioprinting has the great advantage of enabling a fast deposition of high viscous cell-loaded hydrogel with reasonable precision. Compromises between high shape fidelity and cell viability, as well as short process times often require many iterations of optimizing process parameters and varying compositions of the hydrogel. To limit the multitude of parameters during extrusion-based bioprinting, a method for rapid process assessment was developed. This enables to define limits for printing temperature, flow rate and nozzle size from basic rheological measurements with regard to the biological and mechanical requirements. The combination of extrusion-based bioprinting with DoD bioprinting allows for precise deposition of low viscous cell suspension and adjustable concentrations of crosslinking agent. Together, the technologies were used to print a bone replacement model by using the predefined process parameters. Adiposed-derived stem cells (ASC) prone to osteogenic differentiation were homogenously extruded in a cuboid structure of 10x10x5 mm. Human umbilical vein endothelial cells (HUVEC) were printed as highly dense cell suspension lines inside the extruded hydrogel to allow a potential vascularization of the structure in vivo.
• F. Koch, M. Wehrle, K. Tröndle, P. Koltay, G. Finkenzeller, R. Zengerle, S. Zimmermann
  Rapid assessment of extrusion based bioprinting by controlling shear stresses on cells
  2019 Transducers 2019, Berlin, 23.06. - 27.06.2019

2018

• K. Tröndle, S. Kartmann, L. Gutzweiler, R. Zengerle, P. Koltay, S. Zimmermann
  Bioprinting with spheroids: Automated large-scale production and deposition
  2018 3D Cell Culture 2018, 5. - 7. Juni 2018, Freiburg
• K. Tröndle
  Drop-on-Demand Bioprinting Process for Single-Spheroid Deposition and Immobilization
  2018 International Conference on Biofabrication, Würzburg, 28. – 31.10.2018