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Open AccessArticle

Image-Based Histological Evaluation of Scaffold-Free 3D Osteoblast Cultures

Department of Orthopaedics and Sportsorthopaedics, Klinikum rechts der Isar, Technical University of Munich, Ismaninger Strasse 22, 81675 Munich, Germany
Chair of Non-Destructive Testing, Centre for Building Materials, Technical University of Munich, Baumbachstrasse 7, 81245 Munich, Germany
Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove 4059, Australia
Author to whom correspondence should be addressed.
J. Funct. Morphol. Kinesiol. 2017, 2(4), 42;
Received: 31 October 2017 / Revised: 20 November 2017 / Accepted: 20 November 2017 / Published: 24 November 2017
The analysis of tissue network characteristics and cell distribution using histological methods is widely used. However, image analysis still relies on manual evaluation methods, known as semi-quantitative analysis, which are time-consuming and to a certain degree user-specific. For this reason, automated imaging processing methods have an enormous potential to increase sample processing and reduce the variation that is caused by a user-specific evaluation. This work demonstrates the feasibility of using a semi-automated image analysis process based on the open source software framework ImageJ and the plug-in Angiogenesis Analyzer to evaluate the quantitative degree of tissue damage within 3D cell constructs after mechanical loading. Within a proof-of-concept study, the semi-automated approach was applied to calculate the Node-to-Free Ratio (N/F-Ratio) and perform a strut analysis for histological evaluation of mechanically compressed samples of human osteoblast-derived 3D constructs. The N/F-Ratio revealed a median value of 1.29 for the control, whereas the values for the mechanically compressed samples decreased to 0.97 for 20% compression, 0.85 for 40%, and 0.86 for 60%. The strut analysis indicated a decrease of the connected branches with increasing compression rate. The newly developed and time-saving processing workflow was successfully established and can be carried out using available, open source software solutions. View Full-Text
Keywords: bone tissue engineering; 3D cell culture; histomorphometry; open-source software; image-based analysis; ImageJ bone tissue engineering; 3D cell culture; histomorphometry; open-source software; image-based analysis; ImageJ
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MDPI and ACS Style

Eggert, S.; Tuebel, J.; Foehr, P.; Kuntz, L.; Obermeier, A.; Marthen, C.; Grosse, C.U.; Burgkart, R. Image-Based Histological Evaluation of Scaffold-Free 3D Osteoblast Cultures. J. Funct. Morphol. Kinesiol. 2017, 2, 42.

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