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Nanomechanical Analysis of Extracellular Matrix and Cells in Multicellular Spheroids

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Abstract

Introduction

Over the last decade, atomic force microscopy (AFM) has played an important role in understanding nanomechanical properties of various cancer cell lines. This study is focused on Lewis lung carcinoma cell tumours as 3D multicellular spheroid (MS). Not much is know about the mechanical properties of the cells and the surrounding extracellular matrix (ECM) in rapidly growing tumours.

Methods

Depth-dependent indentation measurements were conducted with the AFM. Force-vs.-indentation curves were used to create stiffness profiles as a function of depth. Here studies were focused on the outer most layer, i.e., proliferation zone of the spheroid.

Results

Both surface and sub-surface stiffness profiles of MS were created. This study revealed three nanomechanical topographies, Type A-high modulus due to collagen fibers, Type B-high stiffness at cell membrane and ECM interface and Type C-increased modulus due to cell lying deep inside matrix at a depth of 1.35 μm. Both Type and Type-B topographies result from collagen-based structures in ECM.

Conclusion

This study has first time revealed mechanical constitution of an MS. Depth-dependent indentation studies have the revealed role of various molecular and cellular components responsible for providing mechanical stability to MS. Nanomechanical heterogeneities revealed in this investigation can shed new light in developing correct dosage regime for collagenase treatment of tumours and designing better controlled artificial extracellular matrix systems for replicating tissue growth in-vitro.

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Author Contributions

Conceived and designed the experiments: VV BDH. Performed the experiments: VV. Analyzed the data: VV BDH. Contributed biological material & cells: MS GGMD. Wrote the paper: VV.

Conflict of interest

Varun Vyas, Melani Solomon, Gerard G.M. D’Souza and Bryan D. Huey declare that they have no conflicts of interest.

Ethical Approval

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

Funding

Provided by the University of Connecticut’s Provosts Fund and NSF Nano-Bio-Mechanics grant 0626231.

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Authors and Affiliations

  1. Materials Science and Engineering Department, University of Connecticut, 97 North Eagleville Road, Unit 3136, Storrs, CT, 06269-3136, USA

    Varun Vyas & Bryan D. Huey

  2. Pharmaceutical Sciences, Massachusetts College of Pharmacy & Health Sciences, Boston, MA, 02115, USA

    Melani Solomon & Gerard G. M. D’Souza

  3. McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, MD, 21205, USA

    Melani Solomon

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  1. Varun Vyas
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  2. Melani Solomon
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Correspondence to Varun Vyas.

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Vyas, V., Solomon, M., D’Souza, G.G.M. et al. Nanomechanical Analysis of Extracellular Matrix and Cells in Multicellular Spheroids. Cel. Mol. Bioeng. 12, 203–214 (2019). https://doi.org/10.1007/s12195-019-00577-0

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