Force spectroscopy reveals membrane fluctuations and surface adhesion of extracellular nanovesicles impact their elastic behavior
Artikel i vetenskaplig tidskrift, 2025

The elastic properties of nanoscale extracellular vesicles (EVs) are believed to influence their cellular interactions, thus having a profound implication in intercellular communication. However, accurate quantification of their elastic modulus is challenging due to their nanoscale dimensions and their fluid-like lipid bilayer. We show that the previous attempts to develop atomic force microscopy-based protocol are flawed as they lack theoretical underpinning as well as ignore important contributions arising from the surface adhesion forces and membrane fluctuations. We develop a protocol comprising a theoretical framework, experimental technique, and statistical approach to accurately quantify the bending and elastic modulus of EVs. The method reveals that membrane fluctuations play a dominant role even for a single EV. The method is then applied to EVs derived from human embryonic kidney cells and their genetically engineered classes altering the tetraspanin expression. The data show a large spread; the area modulus is in the range of 4 to 19 mN/m and the bending modulus is in the range of 15 to 33 kBT, respectively. Surprisingly, data for a single EV, revealed by repeated measurements, also show a spread that is attributed to their compositionally heterogeneous fluid membrane and thermal effects. Our protocol uncovers the influence of membrane protein alterations on the elastic modulus of EVs.

elasticity

lipid bilayer

force spectroscopy

extracellular vesicles

atomic force microscopy

Författare

Fredrik Stridfeldt

Kungliga Tekniska Högskolan (KTH)

Vikash Pandey

Stockholms universitet

Hanna Kylhammar

Kungliga Tekniska Högskolan (KTH)

Moein Talebian Gevari

Uppsala universitet

Prattakorn Metem

Kungliga Tekniska Högskolan (KTH)

Vipin Agrawal

Stockholms universitet

Robert R. McCormick School of Engineering and Applied Science

André Görgens

Karolinska Institutet

Karolinska universitetssjukhuset

Universitats Klinikum Essen und Medizinische Fakultat

Doste R. Mamand

Karolinska Institutet

Karolinska universitetssjukhuset

Jennifer Gilbert

Chalmers, Life sciences, Kemisk biologi

Lukas Palmgren

Student vid Chalmers

Margaret Holme

Chalmers, Life sciences, Kemisk biologi

Oskar Gustafsson

Karolinska universitetssjukhuset

Karolinska Institutet

Samir El Andaloussi

Karolinska universitetssjukhuset

Karolinska Institutet

Dhrubaditya Mitra

Stockholms universitet

Apurba Dev

Uppsala universitet

Kungliga Tekniska Högskolan (KTH)

Proceedings of the National Academy of Sciences of the United States of America

0027-8424 (ISSN) 1091-6490 (eISSN)

Vol. 122 16 e2414174122

Ämneskategorier (SSIF 2025)

Den kondenserade materiens fysik

Biofysik

DOI

10.1073/pnas.2414174122

Relaterade dataset

Atomic Force Microscopy (AFM) Data of Exosomes and Liposomes [dataset]

DOI: https://doi.org/10.5281/zenodo.15113782

Mer information

Senast uppdaterat

2025-05-08