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Physical understanding of axonal growth patterns on grooved substrates: groove ridge crossing versus longitudinal alignment

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Abstract

Surface topographies such as micrometric edges and grooves have been widely used to improve neuron outgrowth. However, finding the mechanism of neuron–surface interactions on grooved substrates remains a challenge. In this work, PC12 cells and chick forebrain neurons (CFNs) were cultured on grooved and smooth polyacrylonitrile substrates. It was found that CFNs showed a tendency of growing across groove ridges; while PC12 cells were only observed to grow in the longitudinal direction of grooves. To further investigate these observations, a 3D physical model of axonal outgrowth was developed. In this model, axon shafts are simulated as elastic 3D beams, accounting for the axon outgrowth as well as the focal contacts between axons and substrates. Moreover, the bending direction of axon tips during groove ridge crossing is governed by the energy minimization principle. Our physical model predicts that axonal groove ridge crossing is contributed by the bending compliance of axons, caused by lower Young’s modulus and smaller diameters. This work will aid the understanding of the mechanisms involved in axonal alignment and elongation of neurons guided by grooved substrates, and the obtained insights can be used to enhance the design of instructive scaffolds for nerve tissue engineering and regeneration applications.

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Acknowledgements

This study was partially supported by the National Key Research and Development Program of China (Grant No. 2018YFA0703000), the Key Research and Development Program of Zhejiang Province (Grant No. 2017C01063), the Science Fund for Creative Research Groups of the National Natural Science Foundation of China (Grant No. 51821093), the National Natural Science Foundation of China (Grant Nos. U1609207, 11672268), ZJU Scholarship for Outstanding Doctoral Candidates and Scholarship Program supported by China Scholarship Council (No. 201906320187).

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Authors and Affiliations

  1. The State Key Laboratory of Fluid Power and Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, Hangzhou, 310028, China

    Deming Zhang, Jun Yin & Jianzhong Fu

  2. Key Laboratory of 3D Printing Process and Equipment of Zhejiang Province, School of Mechanical Engineering, Zhejiang University, Hangzhou, 310028, China

    Deming Zhang, Jun Yin & Jianzhong Fu

  3. School of Automation, Hangzhou Dianzi University, Hangzhou, 310018, China

    Hairui Suo

  4. Key Laboratory of Soft Machines and Smart Devices of Zhejiang Province, Department of Engineering Mechanics, Zhejiang University, Hangzhou, 310027, China

    Jin Qian

  5. Department of Mechanical and Aerospace Engineering, University of Florida, Gainesville, FL, 32611, USA

    Yong Huang

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  1. Deming Zhang
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  2. Hairui Suo
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  3. Jin Qian
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  4. Jun Yin
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Contributions

Deming Zhang carried out the numerical simulations and drafted the manuscript. Hairui Suo performed in vitro cell outgrowth and experimental data analysis. Jin Qian worked on the nanoindentation tests and revised the manuscript. Jun Yin is the principal investigator, designed the numerical model, organized, and revised the manuscript. Yong Huang and Jianzhong Fu helped to revise the manuscript, and coordinate the study.

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Correspondence to Jun Yin.

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Deming Zhang, Hairui Suo, Jin Qian, Jun Yin, Jianzhong Fu, and Yong Huang declare that they have no conflict of interest.

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This article does not contain any studies with human or animal subjects performed by any of the authors.

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Zhang, D., Suo, H., Qian, J. et al. Physical understanding of axonal growth patterns on grooved substrates: groove ridge crossing versus longitudinal alignment. Bio-des. Manuf. 3, 348–360 (2020). https://doi.org/10.1007/s42242-020-00089-1

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