Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures
Abstract Three-dimensional (3D) cell cultures are to date the gold standard in biomedical research fields due to their enhanced biological functions compared to conventional two-dimensional (2D) cultures. 3D cell spheroids, as well as organoids, are better suited to replicate tissue functions, which...
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BMC
2023-12-01
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| Series: | Journal of Biological Engineering |
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| Online Access: | https://doi.org/10.1186/s13036-023-00395-z |
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| author | Michael Killinger Adéla Kratochvilová Eva Ingeborg Reihs Eva Matalová Karel Klepárník Mario Rothbauer |
| author_facet | Michael Killinger Adéla Kratochvilová Eva Ingeborg Reihs Eva Matalová Karel Klepárník Mario Rothbauer |
| author_sort | Michael Killinger |
| collection | DOAJ |
| description | Abstract Three-dimensional (3D) cell cultures are to date the gold standard in biomedical research fields due to their enhanced biological functions compared to conventional two-dimensional (2D) cultures. 3D cell spheroids, as well as organoids, are better suited to replicate tissue functions, which enables their use both as in vitro models for basic research and toxicology, as well as building blocks used in tissue/organ biofabrication approaches. Culturing 3D spheroids from bone-derived cells is an emerging technology for both disease modelling and drug screening applications. Bone tissue models are mainly limited by the implementation of sophisticated devices and procedures that can foster a tissue-specific 3D cell microenvironment along with a dynamic cultivation regime. In this study, we consequently developed, optimized and characterized an advanced perfused microfluidic platform to improve the reliability of 3D bone cell cultivation and to enhance aspects of bone tissue maturation in vitro. Moreover, biomechanical stimulation generated by fluid flow inside the arrayed chamber, was used to mimic a more dynamic cell environment emulating a highly vascularized bone we expected to improve the osteogenic 3D microenvironment in the developed multifunctional spheroid-array platform. The optimized 3D cell culture protocols in our murine bone-on-a-chip spheroid model exhibited increased mineralization and viability compared to static conditions. As a proof-of-concept, we successfully confirmed on the beneficial effects of a dynamic culture environment on osteogenesis and used our platform for analysis of bone-derived spheroids produced from primary human pre-osteoblasts. To conclude, the newly developed system represents a powerful tool for studying human bone patho/physiology in vitro under more relevant and dynamic culture conditions converging the advantages of microfluidic platforms with multi-spheroid array technologies. Graphical Abstract |
| format | Article |
| id | doaj-art-c8237eb117c14b73b8ec81e41dc5544a |
| institution | DOAJ |
| issn | 1754-1611 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Biological Engineering |
| spelling | doaj-art-c8237eb117c14b73b8ec81e41dc5544a2025-08-20T03:18:28ZengBMCJournal of Biological Engineering1754-16112023-12-0117111710.1186/s13036-023-00395-zMicrofluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell culturesMichael Killinger0Adéla Kratochvilová1Eva Ingeborg Reihs2Eva Matalová3Karel Klepárník4Mario Rothbauer5Department of Bioanalytical Instrumentation, Institute of Analytical Chemistry, Academy of SciencesLaboratory of Odontogenesis and Osteogenesis, Institute of Animal Physiology and Genetics, Academy of SciencesCell Chip Group, Institute of Applied Synthetic Chemistry, Institute of Chemical Technologies and Analytics, Faculty of Technical Chemistry, Technical University ViennaLaboratory of Odontogenesis and Osteogenesis, Institute of Animal Physiology and Genetics, Academy of SciencesDepartment of Bioanalytical Instrumentation, Institute of Analytical Chemistry, Academy of SciencesCell Chip Group, Institute of Applied Synthetic Chemistry, Institute of Chemical Technologies and Analytics, Faculty of Technical Chemistry, Technical University ViennaAbstract Three-dimensional (3D) cell cultures are to date the gold standard in biomedical research fields due to their enhanced biological functions compared to conventional two-dimensional (2D) cultures. 3D cell spheroids, as well as organoids, are better suited to replicate tissue functions, which enables their use both as in vitro models for basic research and toxicology, as well as building blocks used in tissue/organ biofabrication approaches. Culturing 3D spheroids from bone-derived cells is an emerging technology for both disease modelling and drug screening applications. Bone tissue models are mainly limited by the implementation of sophisticated devices and procedures that can foster a tissue-specific 3D cell microenvironment along with a dynamic cultivation regime. In this study, we consequently developed, optimized and characterized an advanced perfused microfluidic platform to improve the reliability of 3D bone cell cultivation and to enhance aspects of bone tissue maturation in vitro. Moreover, biomechanical stimulation generated by fluid flow inside the arrayed chamber, was used to mimic a more dynamic cell environment emulating a highly vascularized bone we expected to improve the osteogenic 3D microenvironment in the developed multifunctional spheroid-array platform. The optimized 3D cell culture protocols in our murine bone-on-a-chip spheroid model exhibited increased mineralization and viability compared to static conditions. As a proof-of-concept, we successfully confirmed on the beneficial effects of a dynamic culture environment on osteogenesis and used our platform for analysis of bone-derived spheroids produced from primary human pre-osteoblasts. To conclude, the newly developed system represents a powerful tool for studying human bone patho/physiology in vitro under more relevant and dynamic culture conditions converging the advantages of microfluidic platforms with multi-spheroid array technologies. Graphical Abstracthttps://doi.org/10.1186/s13036-023-00395-zBone-on-a-chip3D cell culturesDynamic cultivationMicrofluidicsMicrowells micropillars |
| spellingShingle | Michael Killinger Adéla Kratochvilová Eva Ingeborg Reihs Eva Matalová Karel Klepárník Mario Rothbauer Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures Journal of Biological Engineering Bone-on-a-chip 3D cell cultures Dynamic cultivation Microfluidics Microwells micropillars |
| title | Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures |
| title_full | Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures |
| title_fullStr | Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures |
| title_full_unstemmed | Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures |
| title_short | Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures |
| title_sort | microfluidic device for enhancement and analysis of osteoblast differentiation in three dimensional cell cultures |
| topic | Bone-on-a-chip 3D cell cultures Dynamic cultivation Microfluidics Microwells micropillars |
| url | https://doi.org/10.1186/s13036-023-00395-z |
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