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Osteogenic cells differentiation on topological surfaces under ultrasound stimulation

  • Materials for life sciences
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Abstract

The current trends in bone tissue engineering aim to fasten the cells osteogenic differentiation by mechanical stimulation. To date, several approaches have proved efficient for this purpose. One is related to changing the shape of the cells nuclei using topological surfaces with appropriate dimensions and stiffness. Another successful method is by low-intensity pulsed ultrasound stimulation (LIPUS) of the cells. The goal of this proof-of-concept study is to introduce and validate, for the first time, the synergistic effect of topological surfaces and LIPUS for improving the osteogenic differentiation of osteoblast-like cells. Cells were grown on topological surfaces consisting of vertical microtubes fabricated by laser direct writing. The flexibility of the topological surfaces was tuned by varying the microtubes’ height. The spatial arrangement and dimensions of the microtubes limited the cell–cell interactions and allowed us to observe individual cells. A finite element model simulation was proposed for explaining the cell–surface interaction details. We monitored the cells nuclei deformations in response to the topological surfaces in conjunction with LIPUS. The topological surfaces alone induced dramatic changes of the shape of the cells nuclei that wrapped around the microtubes. The nuclei deformation was further increased by LIPUS. This synergy between the topological surfaces and LIPUS allowed us to obtain an increase of up to 200% in the cells osteogenic differentiation, as determined by ALP activity and osteocalcin secretion measurements, in comparison with flat surfaces in static regime. A causal relationship between the nuclei deformation and the cells osteogenic differentiation was established.

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Acknowledgements

This work was supported by a grant of the Romanian National Authority for Scientific Research and Innovation, CNCS/CCCDI—UEFISCDI, project number PN-III-P2-2.1-PED-2016-1787. A part of this work was supported by the National Program LAPLAS VI (16N/08.02.2019).

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Authors and Affiliations

  1. Center for Advanced Laser Technologies (CETAL), National Institute for Laser, Plasma and Radiation Physics, 077125, Magurele, Romania

    Irina Alexandra Paun, Bogdan Stefanita Calin & Catalin Romeo Luculescu

  2. Faculty of Applied Sciences, University Politehnica of Bucharest, 060042, Bucharest, Romania

    Irina Alexandra Paun, Bogdan Stefanita Calin, Cosmin Catalin Mustaciosu & Mona Mihailescu

  3. Horia Hulubei National Institute for Physics and Nuclear Engineering IFIN-HH, 077125, Magurele, Romania

    Cosmin Catalin Mustaciosu

  4. Faculty of Medical Engineering, University Politehnica of Bucharest, 011061, Bucharest, Romania

    Cezar Stefan Popovici

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  1. Irina Alexandra Paun
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Correspondence to Irina Alexandra Paun.

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10853_2019_3680_MOESM1_ESM.docx

We provide supplementary information concerning: 1) analysis of the fluorescence images of the cells nuclei using a code built in MATLAB; 2) Finite Element Model simulation performed in SolidWorks® that evaluates the microtubes’ deformation at the contact with the cells; 3) normal vibration modes of the microtubes and of the cells on microtubes; 4) scanning electron micrographs of topological surfaces fabricated by laser direct writing via two photon-polymerization of IP-L780 photopolymer, with microtubes heights above 20 μm; 5) wide area scanning electron micrograph of topological surface fabricated by laser direct writing via two-photon polymerization of IP-L780 photopolymer, for microtubes height of 20 μm (DOCX 6887 kb)

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Paun, I.A., Calin, B.S., Mustaciosu, C.C. et al. Osteogenic cells differentiation on topological surfaces under ultrasound stimulation. J Mater Sci 54, 11213–11230 (2019). https://doi.org/10.1007/s10853-019-03680-9

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