A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis
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-T...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1167623/full |
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| author | Annachiara Scalzone Annachiara Scalzone Giorgia Cerqueni Xiao Nong Wang Kenny Dalgarno Monica Mattioli-Belmonte Ana M. Ferreira-Duarte Piergiorgio Gentile |
| author_facet | Annachiara Scalzone Annachiara Scalzone Giorgia Cerqueni Xiao Nong Wang Kenny Dalgarno Monica Mattioli-Belmonte Ana M. Ferreira-Duarte Piergiorgio Gentile |
| author_sort | Annachiara Scalzone |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-09T13:45:27Z |
| format | Article |
| id | doaj.art-f0e2bdce08b54e4facbd4ed18c96c640 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-09T13:45:27Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-f0e2bdce08b54e4facbd4ed18c96c6402023-05-09T05:08:03ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-05-011110.3389/fbioe.2023.11676231167623A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesisAnnachiara Scalzone0Annachiara Scalzone1Giorgia Cerqueni2Xiao Nong Wang3Kenny Dalgarno4Monica Mattioli-Belmonte5Ana M. Ferreira-Duarte6Piergiorgio Gentile7School of Engineering, Newcastle University, Newcastle UponTyne, United KingdomCenter for Advanced Biomaterials for Healthcare@CRIB Istituto Italiano di Tecnologia, Napoli, ItalyDepartment of Clinical and Molecular Sciences (DISCLIMO), Università Politecnica delle Marche, Ancona, ItalyTranslational and Clinical Research Institute, Newcastle University, Newcastle UponTyne, United KingdomSchool of Engineering, Newcastle University, Newcastle UponTyne, United KingdomDepartment of Clinical and Molecular Sciences (DISCLIMO), Università Politecnica delle Marche, Ancona, ItalySchool of Engineering, Newcastle University, Newcastle UponTyne, United KingdomSchool of Engineering, Newcastle University, Newcastle UponTyne, United KingdomGiven 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.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1167623/fullin vitro modelarticular cartilageosteoarthritiscytokineschondrocytes |
| spellingShingle | Annachiara Scalzone Annachiara Scalzone Giorgia Cerqueni Xiao Nong Wang Kenny Dalgarno Monica Mattioli-Belmonte Ana M. Ferreira-Duarte Piergiorgio Gentile A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis Frontiers in Bioengineering and Biotechnology in vitro model articular cartilage osteoarthritis cytokines chondrocytes |
| title | A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis |
| title_full | A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis |
| title_fullStr | A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis |
| title_full_unstemmed | A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis |
| title_short | A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis |
| title_sort | cytokine induced spheroid based in vitro model for studying osteoarthritis pathogenesis |
| topic | in vitro model articular cartilage osteoarthritis cytokines chondrocytes |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1167623/full |
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