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Positive impact of dynamic seeding of mesenchymal stem cells on bone-like biodegradable scaffolds with increased content of calcium phosphate nanoparticles

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Abstract

One of the main aims of bone tissue engineering, regenerative medicine and cell therapy is development of an optimal artificial environment (scaffold) that can trigger a favorable response within the host tissue, it is well colonized by resident cells of organism and ideally, it can be in vitro pre-colonized by cells of interest to intensify the process of tissue regeneration. The aim of this study was to develop an effective tool for regenerative medicine, which combines the optimal bone-like scaffold and colonization technique suitable for cell application. Accordingly, this study includes material (physical, chemical and structural) and in vitro biological evaluation of scaffolds prior to in vivo study. Thus, porosity, permeability or elasticity of two types of bone-like scaffolds differing in the ratio of collagen type I and natural calcium phosphate nanoparticles (bCaP) were determined, then analyzes of scaffold interaction with mesenchymal stem cells (MSCs) were performed. Simultaneously, dynamic seeding using a perfusion bioreactor followed by static cultivation was compared with standard static cultivation for the whole period of cultivation. In summary, cell colonization ability was estimated by determination of cell distribution within the scaffold (number, depth and homogeneity), matrix metalloproteinase activity and gene expression analysis of signaling molecules and differentiation markers. Results showed, the used dynamic colonization technique together with the newly-developed collagen-based scaffold with high content of bCaP to be an effective combined tool for producing bone grafts for bone implantology and regenerative medicine.

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Acknowledgements

This study was supported by project 15-25813A awarded by the Ministry of Health of the Czech Republic and PROGRES Q26 provided by Charles University, Czech Republic. Special thanks go to Blanka Bilkova for her technical assistance.

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Authors and Affiliations

  1. Institute of Pathological Physiology, 1st Faculty of Medicine, Charles University, U nemocnice 5, Prague, 128 53, Czech Republic

    Pavla Sauerova, Martin Bartos & Marie Hubalek Kalbacova

  2. Biomedical Centre, Faculty of Medicine in Pilsen, Charles University, Alej Svobody 1655/76, Pilsen, 323 00, Czech Republic

    Pavla Sauerova, Tereza Kubikova, Zbynek Tonar & Marie Hubalek Kalbacova

  3. Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University, Karlovarská 48, Plzeň, 301 00, Czech Republic

    Tereza Kubikova & Zbynek Tonar

  4. Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, V Holesovickach 41, Prague, 182 09, Czech Republic

    Tomas Suchy & Monika Supova

  5. Laboratory of Biomechanics, Department of Mechanics, Biomechanics and Mechatronics, Faculty of Mechanical Engineering, Czech Technical University in Prague, Technicka 4, Prague, 166 07, Czech Republic

    Tomas Suchy & Radek Sedlacek

  6. Institute of Dental Medicine, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, Katerinska 32, Prague, 121 08, Czech Republic

    Martin Bartos

  7. Laboratory of Cell Regeneration and Plasticity, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Rumburska 89, Libechov, 277 21, Czech Republic

    Jiri Klima, Jana Juhasova & Stefan Juhas

  8. Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 201 33, Milan, Italy

    Marco Piola, Gianfranco Beniamino Fiore & Monica Soncini

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  1. Pavla Sauerova
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  2. Tomas Suchy
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Correspondence to Marie Hubalek Kalbacova.

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All experiments were carried out according to the guidelines for the care and use of experimental animals and approved by the Resort Professional Commission of the Czech Academy of Sciences for Approval of Projects of Experiments on Animals (Approved protocol No 32/2015 and 53/2015).

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Sauerova, P., Suchy, T., Supova, M. et al. Positive impact of dynamic seeding of mesenchymal stem cells on bone-like biodegradable scaffolds with increased content of calcium phosphate nanoparticles. Mol Biol Rep 46, 4483–4500 (2019). https://doi.org/10.1007/s11033-019-04903-7

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