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ORIGINAL RESEARCH article

Front. Bioeng. Biotechnol.
Sec. Tissue Engineering and Regenerative Medicine
Volume 12 - 2024 | doi: 10.3389/fbioe.2024.1416694
This article is part of the Research Topic A New Age for Articular Cartilage: from Bench to Beside of Tissue Engineering and Regenerative Medicine in Cartilage Tissue Repair View all 4 articles

Influence of inflammation on the expression of microRNA-140 in extracellular vesicles from 2D and 3D culture models of synovial membrane-derived stem cells

Provisionally accepted
João Pedro H. Pfeifer João Pedro H. Pfeifer 1,2*Fernanda d. Stievani Fernanda d. Stievani 1Célio J. Fernandes Célio J. Fernandes 3Gustavo Rosa Gustavo Rosa 1Emanuel Vitor P. Apolonio Emanuel Vitor P. Apolonio 1Mariana C. Rossi Mariana C. Rossi 1Willian Zambuzzi Willian Zambuzzi 4Ana Liz G. Alves Ana Liz G. Alves 1
  • 1 School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, Brazil
  • 2 São Leopoldo Mandic School, Campinas, Brazil
  • 3 Department of Physics and Biophysics, Institute of Biosciences, Sao Paulo State University, Botucatu, Sao Paulo, Brazil
  • 4 Department of Chemical and Biological Sciences, Institute of Biosciences, São Paulo State University, Botucatu, Brazil

The final, formatted version of the article will be published soon.

    Background: In osteoarthritis (OA), articular homeostasis is regulated, for instance, by microRNA-140, which inhibits ADAMTS-5 an enzyme that cleaves aggrecan and 2 stimulates the synthesis of other inflammatory mediators. This study aims to evaluate the expression of microRNA-140 in extracellular vesicles (EVs) derived from equine synovial membrane-derived MSCs (eqSMMSCs) cultured in monolayer (2D) and three-dimensional (3D) culture models under an in vitro inflammatory environment.Methods: Four experimental groups of eqSMMSC cultures were defined for EVs isolation. Control groups 2D and 3D were cultured in conventional cell culture medium. Treated groups 2D-OA and 3D-OA were exposed to an osteoarthritis-like medium containing IL-1β and TNFα. Culture media samples were collected at 24h, 72h, and 120h time points for EV isolation and characterized using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). RT-qPCR was employed to assess the expression of microRNA-140 in both cells and EVs. All statistical analyses were conducted at a significance level of 5%.Results: Encapsulation of eqSMMSCs protected cells from the inflammatory medium compared to monolayer culture. EVs were found in higher concentrations in 3D-OA cultures. Additionally, higher expression of miRNA-140 was observed in cells of the 3D-OA group at 24 and 72 hours, whereas miRNA-140 expression in EVs was higher in the 3D group at 72 hours and in the 2D-OA group at 120 hours (P < 0.001).However, 3D-OA cultures showed higher expression of mRNA Adamts5 in EVs at 120 hours.The response of eqSMMSCs to inflammatory stimuli involves intracellular expression of miRNA-140 and its subsequent transport via EVs, with a quicker response observed in 3D cultures compared to 2D cultures. This study shed light into the behavior of stem cells in restoring homeostasis in osteoarthritic joints.

    Keywords: articular homeostasis, Equine, Exosomes, microRNA, Osteoarthritis, Nanotechnology, translational medicine

    Received: 12 Apr 2024; Accepted: 17 Jul 2024.

    Copyright: © 2024 Pfeifer, Stievani, Fernandes, Rosa, Apolonio, Rossi, Zambuzzi and Alves. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: João Pedro H. Pfeifer, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, Brazil

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