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A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis

Lookup NU author(s): Dr Annachiara ScalzoneORCiD, Dr Xiao WangORCiD, Professor Kenneth Dalgarno, Dr Ana Ferreira-DuarteORCiD, Dr Piergiorgio GentileORCiD



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Copyright © 2023 Scalzone, Cerqueni, Wang, Dalgarno, Mattioli-Belmonte, Ferreira-Duarte and Gentile.Given the lack of in vitro models faithfully reproducing the osteoarthritis (OA) disease on-set, this work aimed at manufacturing a reliable and predictive in vitro cytokine-based Articular Cartilage (AC) model to study OA progression. Cell spheroids of primary human fetal chondrocytes (FCs) and h-TERT mesenchymal stem cells differentiated chondrocytes (Y201-C) were analysed in terms of growth kinetics, cells proliferation and apoptosis over 10 days of culture, in healthy condition or in presence of cytokines (interleukin-1ß, −6 and TNF-α). Then, the spheroids were assembled into chondrospheres using a bottom-up strategy, to obtain an in vitro cytokines-induced OA model. The resulting chondrospheres were evaluated for gene expression and anabolic ECM proteins. Compared to the healthy environment, the simulated OA environment induced chondrocyte hyperproliferation and apoptotic pathway, decreased expression of anabolic ECM proteins, and diminished biosynthetic activity, resembling features of early-stage OA. These characteristics were observed for both Y201-C and HC at high and low concentrations of cytokines. Both HC and Y201-C demonstrated the suitability for the manufacturing of a scaffold-free in vitro OA model to facilitate studies into OA pathogenesis and therapeutic strategies. Our approach provides a faithful reproduction of early-stage osteoarthritis, demonstrating the ability of obtaining different disease severity by tuning the concentration of OA-related cytokines. Given the advantages in easy access and more reproducible performance, Y201-C may represent a more favourable source of chondrocytes for establishing more standardized protocols to obtain OA models.

Publication metadata

Author(s): Scalzone A, Cerqueni G, Wang XN, Dalgarno K, Mattioli-Belmonte M, Ferreira-Duarte AM, Gentile P

Publication type: Article

Publication status: Published

Journal: Frontiers in Bioengineering and Biotechnology

Year: 2023

Volume: 11

Online publication date: 09/05/2023

Acceptance date: 27/04/2023

Date deposited: 06/06/2023

ISSN (electronic): 2296-4185

Publisher: Frontiers Media S.A.


DOI: 10.3389/fbioe.2023.1167623


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Funder referenceFunder name
21156VERSUS Arthritis (formerly Arthritis Research UK)