Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation

Osteoarthritis (OA) is a musculoskeletal degenerative disease characterized by alterations in cartilage and subchondral bone leading to impaired joint function. OA disproportionally affects females more than males, yet the molecular mechanisms underlying these biological sex differences remain elusi...

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Main Authors: Francisco Conceição, João Meneses, Filipa Lebre, Malin Becker, Nuno Araújo-Gomes, Rianne Vos, Ana R. Ribeiro, Ernesto Alfaro-Moreno, Jeroen Leijten, Liliana Moreira Teixeira
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Language:English
Published: Elsevier 2025-06-01
Series:Materials Today Bio
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Online Access:http://www.sciencedirect.com/science/article/pii/S259000642500287X
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author Francisco Conceição
João Meneses
Filipa Lebre
Malin Becker
Nuno Araújo-Gomes
Rianne Vos
Ana R. Ribeiro
Ernesto Alfaro-Moreno
Jeroen Leijten
Liliana Moreira Teixeira
author_facet Francisco Conceição
João Meneses
Filipa Lebre
Malin Becker
Nuno Araújo-Gomes
Rianne Vos
Ana R. Ribeiro
Ernesto Alfaro-Moreno
Jeroen Leijten
Liliana Moreira Teixeira
author_sort Francisco Conceição
collection DOAJ
description Osteoarthritis (OA) is a musculoskeletal degenerative disease characterized by alterations in cartilage and subchondral bone leading to impaired joint function. OA disproportionally affects females more than males, yet the molecular mechanisms underlying these biological sex differences remain elusive. Current therapeutic strategies to halt the progression of OA are still lacking, in part due to the limited predictive potential of standard models which often do not account for sex disparities. Herein, an organ-on-chip microfluidic platform was developed to model the osteochondral unit, composed of adjacent bone and cartilage culture chambers, and capture sex-specific hallmarks of OA. Sex-stratified human primary chondrocytes and osteoblasts were compartmentalized within biomimetic hydrogels emulating the bone-cartilage interface, which were subjected to inflammatory triggers to mimic the onset of OA. We confirmed that interleukin-1β and Tumor Necrosis Factor-α stimulation triggered upregulation of pro-inflammatory cytokines and matrix metalloproteinases related genes in all donors, with marginal trends for increased expression in female cells. In addition, metabolic labeling coupled with confocal imaging revealed that inflammatory stimulation modulated extracellular matrix deposition by human chondrocytes in a sex-specific fashion. Not only matrix deposition but also matrix remodeling was altered upon inflammation, leading to a significant reduction in matrix stiffness in both cartilage and bone compartments. Overall, sex-stratified osteochondral unit on-chips offer novel insights into sex-specific cellular responses to inflammatory insults, demonstrating the importance of incorporating sex stratification in emergent organ-on-chip models. Thus, this platform provides a physiologically relevant 3D microenvironment to further investigate sex-specific drivers of OA, paving the way for targeted therapies.
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spelling doaj-art-a1361f21eaff441ba2bcc0d9beeda67a2025-08-20T01:54:15ZengElsevierMaterials Today Bio2590-00642025-06-013210172810.1016/j.mtbio.2025.101728Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulationFrancisco Conceição0João Meneses1Filipa Lebre2Malin Becker3Nuno Araújo-Gomes4Rianne Vos5Ana R. Ribeiro6Ernesto Alfaro-Moreno7Jeroen Leijten8Liliana Moreira Teixeira9Department of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the NetherlandsDepartment of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the NetherlandsNanosafety Group, International Iberian Nanotechnology Laboratory, 4715-330, Braga, PortugalOptics11 Life, 1101 BM, Amsterdam, the NetherlandsDepartment of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the NetherlandsDepartment of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the NetherlandsNanosafety Group, International Iberian Nanotechnology Laboratory, 4715-330, Braga, PortugalNanosafety Group, International Iberian Nanotechnology Laboratory, 4715-330, Braga, PortugalDepartment of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the NetherlandsDepartment of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the Netherlands; Organ-on-Chip Centre Twente, TechMed Centre, MESA+, University of Twente, 7522 NB, Enschede, the Netherlands; Corresponding author. Department of Bioengineering Technologies, Faculty of Science and Technology, TechMedCentre, University of Twente, 7522 NB, Enschede, the Netherlands.Osteoarthritis (OA) is a musculoskeletal degenerative disease characterized by alterations in cartilage and subchondral bone leading to impaired joint function. OA disproportionally affects females more than males, yet the molecular mechanisms underlying these biological sex differences remain elusive. Current therapeutic strategies to halt the progression of OA are still lacking, in part due to the limited predictive potential of standard models which often do not account for sex disparities. Herein, an organ-on-chip microfluidic platform was developed to model the osteochondral unit, composed of adjacent bone and cartilage culture chambers, and capture sex-specific hallmarks of OA. Sex-stratified human primary chondrocytes and osteoblasts were compartmentalized within biomimetic hydrogels emulating the bone-cartilage interface, which were subjected to inflammatory triggers to mimic the onset of OA. We confirmed that interleukin-1β and Tumor Necrosis Factor-α stimulation triggered upregulation of pro-inflammatory cytokines and matrix metalloproteinases related genes in all donors, with marginal trends for increased expression in female cells. In addition, metabolic labeling coupled with confocal imaging revealed that inflammatory stimulation modulated extracellular matrix deposition by human chondrocytes in a sex-specific fashion. Not only matrix deposition but also matrix remodeling was altered upon inflammation, leading to a significant reduction in matrix stiffness in both cartilage and bone compartments. Overall, sex-stratified osteochondral unit on-chips offer novel insights into sex-specific cellular responses to inflammatory insults, demonstrating the importance of incorporating sex stratification in emergent organ-on-chip models. Thus, this platform provides a physiologically relevant 3D microenvironment to further investigate sex-specific drivers of OA, paving the way for targeted therapies.http://www.sciencedirect.com/science/article/pii/S259000642500287XOrgan-on-chipOsteoarthritisSex differencesMetabolic labelingChondrocytesInflammation
spellingShingle Francisco Conceição
João Meneses
Filipa Lebre
Malin Becker
Nuno Araújo-Gomes
Rianne Vos
Ana R. Ribeiro
Ernesto Alfaro-Moreno
Jeroen Leijten
Liliana Moreira Teixeira
Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation
Materials Today Bio
Organ-on-chip
Osteoarthritis
Sex differences
Metabolic labeling
Chondrocytes
Inflammation
title Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation
title_full Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation
title_fullStr Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation
title_full_unstemmed Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation
title_short Sex-stratified osteochondral organ-on-chip model reveals sex-specific responses to inflammatory stimulation
title_sort sex stratified osteochondral organ on chip model reveals sex specific responses to inflammatory stimulation
topic Organ-on-chip
Osteoarthritis
Sex differences
Metabolic labeling
Chondrocytes
Inflammation
url http://www.sciencedirect.com/science/article/pii/S259000642500287X
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