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Compartment Specific Mechanical Loading of Bi-Layered Osteochondral Micro-Tissues in a Joint-on-Chip System suggests routes for Osteoarthritis pathogenesis investigation

ABSTRACT: In this work we engineered an osteochondral unit (OCU) on-chip model where osteoarthritis (OA) phenotype is recapitulated through the application of hyper-physiological mechanical loading. After a functional and biological validation of the device, we aimed to better characterize the diversity of the human articular chondrocytes (hACs) population present in our in vitro model and identify subpopulations responsible for the pathological loading response. As a method, we applied single-cell RNA sequencing (scRNA-seq) to hACs from both on-chip single cultures and co-cultures with bone marrow derived mesenchymal stromal cells (bmMSCs), i.e. in osteochondral construct, and either exposed or not to cyclic mechanical loading.

ORGANISM(S): Homo sapiens

PROVIDER: GSE237786 | GEO | 2025/09/17

REPOSITORIES: GEO

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Compartment Specific Mechanical Loading of Bi-Layered Osteochondral Micro-Tissues in a Joint-on-Chip System suggests routes for Osteoarthritis pathogenesis investigation

Project description:Compartment Specific Mechanical Loading of Bi-Layered Osteochondral Micro-Tissues in a Joint-on-Chip System suggests routes for Osteoarthritis pathogenesis investigation

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Project description:Osteochondral tissue engineering poses the challenge of combining both cartilage and bone tissue engineering fundamentals. In this study, a sphere-templating technique was applied to fabricate an integrated bi-layered scaffold based on degradable poly(hydroxyethyl methacrylate) hydrogel. One layer of the integrated scaffold was designed with a single defined, monodispersed pore size of 38 μm and pore surfaces coated with hydroxyapatite particles to promote regrowth of subchondral bone while the second layer had 200 μm pores with surfaces decorated with hyaluronan for articular cartilage regeneration. Mechanical properties of the construct as well as cyto-compatibility of the scaffold and its degradation products were elucidated. To examine the potential of the biphasic scaffold for regeneration of osteochondral tissue the designated cartilage and bone layers of the integrated bi-layered scaffold were seeded with chondrocytes differentiated from human mesenchymal stem cells and primary human mesenchymal stem cells, respectively. Both types of cells were co-cultured within the scaffold in standard medium without soluble growth/differentiation factors over four weeks. The ability of the integrated bi-layered scaffold to support simultaneous matrix deposition and adequate cell growth of two distinct cell lineages in each layer during four weeks of co-culture in vitro in the absence of soluble growth factors was demonstrated.

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Single-cell RNA sequencing for osteochondral fusions

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2025-02-21 | GSE275943 | GEO

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GNAS/PKA pathway promotes aberrant osteochondral differentiation of Gli1+ tendon sheath progenitors

Project description:We have employed a single cell sequencing approach using 10x Genomics scRNAseq to study the identity and fate determination of Gli1+ tendon sheath progenitors in heterotopic ossification(HO) formation following tenotomy. Gli1 is a signature gene for several tissue-specific Mesenchymal stem cells (MSCs). Aberrant osteochondral differentiation of MSCs in soft tissues leads to incorrect tissue regeneration into HO. This study demonstrated a subpopulation of Gli1+ tendon sheath cells that function as tendon stem/progenitors and can initially differentiate into normal or osteochondral tendon-lineage progenitors, with the latter subsequently giving rise to chondrocytes and osteocytes in development of tendon HO.

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Tissue Catabolism and Donor-Specific Dexamethasone Response in a Human Osteochondral Model of Post-Traumatic Osteoarthritis

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The equine carpal osteochondral fragment model

Project description:Synovial fluid was obtained weekly from nine horses throughout a 70-day study period and subjected to small RNA sequencing. Nine skeletally mature Standardbred trotters (seven mares and two geldings, 2.5-7 years old, weighing 397-528 kg) were included in this study. OA was surgically induced in the left middle carpal joint and the right middle carpal joint underwent sham surgery. Under arthroscopic guidance, an osteochondral fragment was created in the left middle carpal joint by chiselling off a fragment on the dorsodistal surface of the radial carpal bone using an 8 mm curved osteotome. The fragment was left adherent to the joint capsule proximally, and an incongruent defect bed of approximately 15 mm in width was created in the exposed subchondral bone between the fragment and radial carpal bone by drilling with an arthroscopic burr. Debris from the procedure was left inside the joint. Sham surgery (arthroscopy alone) was performed in the right middle carpal joint. All portals were sutured using 2-0 propylene (Surgipro, Medtronic Danmark A/S, Copenhagen, Denmark). Both carpi were bandaged with sterile dressings. Bandages were changed at day 2 and removed at day 5. Sutures were removed 10 days after surgery. From 2 weeks following OA induction and onwards, horses were exercised in 2 min of trot (4.4-5.3 m/sec) , 2 min of fast trot/gallop (9 m/sec) and 2 min of trot (4.4-5.3 m/sec) 5 days a week on a treadmill. The horses were euthanized on day 71 or 72 with an overdose of pentobarbital sodium (140 mg/kg, Euthasol Vet, Dechra Veterinary Products, Uldum, Denmark). One horse, Horse 5, was euthanized according to predefined humane endpoints at day 49, as it became lame at the walk.

2023-03-01 | E-MTAB-11840 | biostudies-arrayexpress

Functional performance of a bi-layered chitosan-nano-hydroxyapatite osteochondral scaffold: a pre-clinical <i>in vitro</i> tribological study.

Project description:Osteochondral grafts are used for repair of focal osteochondral lesions. Autologous grafts are the gold standard treatment; however, limited graft availability and donor site morbidity restrict use. Therefore, there is a clinical need for different graft sources/materials which replicate natural cartilage function. Chitosan has been proposed for this application. The aim of this study was to assess the biomechanics and biotribology of a bioresorbable chitosan/chitosan-nano-hydroxyapatite osteochondral construct (OCC), implanted in an in vitro porcine knee experimental simulation model. The OCC implanted in different surgical positions (flush, proud and inverted) was compared to predicate grafts in current clinical use and a positive control consisting of a stainless steel graft implanted proud of the cartilage surface. After 3 h (10 800 cycles) wear simulation under a walking gait, subsidence occurred in all OCC samples irrespective of surgical positioning, but with no apparent loss of material and low meniscus wear. Half the predicate grafts exhibited delamination and scratching of the cartilage surfaces. No graft subsidence occurred in the positive controls but wear and deformation of the meniscus were apparent. Implanting a new chitosan-based OCC either optimally (flush), inverted or proud of the cartilage surface resulted in minimal wear, damage and deformation of the meniscus.

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Mechanical loading and gene expression - Microarray study

Project description:Skeletal integrity in humans and animals is maintained by daily mechanical loading. It has been widely accepted that osteocytes function as mechanosensors. Many biochemical signaling molecules are involved in the response of osteocytes to mechanical stimulation. The aim of this study was to identify genes involved in the translation of mechanical stimuli into bone formation. The four-point bending model was used to induce a single period of mechanical loading (comprising 300 cycles (2 Hz) using a peak magnitude of 60 N) on the right tibia, while the contra lateral left tibia served as control. Six hours after loading, the effects of mechanical loading on gene-expression were determined with microarray analysis. Protein expression of differentially regulated genes was evaluated with immunohistochemistry. Nine genes were found to exhibit a significant differential gene expression in LOAD compared to control. MEPE, Garnl1, V2R2B, and QFG TN1 olfactory receptor were up-regulated, and creatine kinase (muscle form), fibrinogen-B beta-polypeptide, monoamine oxidase A, troponin-C and kinesin light chain-C were down-regulated. Validation with real-time RT-PCR analysis confirmed the up regulation of MEPE and the down-regulation of creatine kinase (muscle form) and troponin-C in the loaded tibia. Immunohistochemistry showed that the increase of MEPE protein expression was already detectable six hours after mechanical loading. In conclusion, these genes probably play a role during translation of mechanical stimuli six hours after mechanical loading. The modulation of MEPE expression may indicate a connection between bone mineralization and bone formation after mechanical stimulation. Two groups: LOAD vs contralateral control and SHAM vs contralateral control (n=5/group)

2013-11-21 | E-GEOD-50707 | biostudies-arrayexpress

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