Abstract
Tissue engineering is a promising technique for the repair of bone defects. An efficient and homogeneous distribution of cell seeding into scaffold is a crucial but challenging step in the technique. Murine bone marrow mesenchymal stem cells were seeded into porous hydroxyapatite scaffolds of two morphologies by three methods: static seeding, semi-dynamic seeding, or dynamic perfusion seeding. Seeding efficiency, survival, distribution, and proliferation were quantitatively evaluated. To investigate the performance of the three seeding methods for larger/thicker scaffolds as well as batch seeding of numerous scaffolds, three scaffolds were stacked to form assemblies, and seeding efficiencies and cell distribution were analyzed. The semi-dynamic seeding and static seeding methods produced significantly higher seeding efficiencies, vitalities, and proliferation than did the dynamic perfusion seeding. On the other hand, the semi-dynamic seeding and dynamic perfusion seeding methods resulted in more homogeneous cell distribution than did the static seeding. For stacked scaffold assemblies, the semi-dynamic seeding method also created superior seeding efficiency and longitudinal cell distribution homogeneity. The semi-dynamic seeding method combines the high seeding efficiency of static seeding and satisfactory distribution homogeneity of dynamic seeding while circumventing their disadvantages. It may contribute to improved outcomes of bone tissue engineering.
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Data Availability Statements
The datasets generated during and analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- HA:
-
Hydroxyapatite
- 3D:
-
Three-dimension
- MTT:
-
3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide
- LDH:
-
Lactate dehydrogenase
- MSCs:
-
Mesenchymal stem cells
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Acknowledgements
This research was funded by National Natural Science Foundation of China (2016YFB0700803), Sichuan Science and Technology Program (2020065, 21MZGC0218), Major Project of Education Department in Sichuan Province (18CZ0019), Startup Program of Southwest Normal University (18Q069, 18Q030), Luzhou-SWMU Strategic Cooperation Program (2017LZXNYD-J35).
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Shi, F., Duan, K., Yang, Z. et al. Improved cell seeding efficiency and cell distribution in porous hydroxyapatite scaffolds by semi-dynamic method. Cell Tissue Bank 23, 313–324 (2022). https://doi.org/10.1007/s10561-021-09945-5
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DOI: https://doi.org/10.1007/s10561-021-09945-5