Development of an <i>Ex Vivo</i> Murine Osteochondral Repair Model

Schaik, Thomas J. A. van; Gaul, Florian; Dorthé, Erik W.; Lee, Emily E.; Grogan, Shawn P.; D’Lima, Darryl D.

doi:10.1177/1947603518809402

Cited by 5 publications

(7 citation statements)

References 48 publications

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“…Defects were generated from bovine stifle joint explants using a sterilized 4 mm trephine drill (Brutsch–Ruegger, Urdorf, CH) to centrally remove a full-thickness circular cylindrical cartilage biopsy in dimensions of 4 mm in diameter and 3 mm in depth, as previously described (Figs. 1 C, 2 A) 24 – 26 . Afterward, the obtained explants were cultured in DMEM–HG containing 10% FCS, 1% penicillin/streptomycin at 37°C, and 5% CO 2 for 24 h to ensure the sterility of the explants.…”

Section: Mscs Culture In Hydrogelmentioning

confidence: 99%

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Cartilage tissue engineering using decellularized biomatrix hydrogel containing TGF-β-loaded alginate microspheres in mechanically loaded bioreactor

Bordbar,

Li,

Lotfibakhshaiesh

et al. 2024

Sci Rep

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Physiochemical tissue inducers and mechanical stimulation are both efficient variables in cartilage tissue fabrication and regeneration. In the presence of biomolecules, decellularized extracellular matrix (ECM) may trigger and enhance stem cell proliferation and differentiation. Here, we investigated the controlled release of transforming growth factor beta (TGF-β1) as an active mediator of mesenchymal stromal cells (MSCs) in a biocompatible scaffold and mechanical stimulation for cartilage tissue engineering. ECM-derived hydrogel with TGF-β1-loaded alginate-based microspheres (MSs) was created to promote human MSC chondrogenic development. Ex vivo explants and a complicated multiaxial loading bioreactor replicated the physiological conditions. Hydrogels with/without MSs and TGF-β1 were highly cytocompatible. MSCs in ECM-derived hydrogel containing TGF-β1/MSs showed comparable chondrogenic gene expression levels as those hydrogels with TGF-β1 added in culture media or those without TGF-β1. However, constructs with TGF-β1 directly added within the hydrogel had inferior properties under unloaded conditions. The ECM-derived hydrogel group including TGF-β1/MSs under loading circumstances formed better cartilage matrix in an ex vivo osteochondral defect than control settings. This study demonstrates that controlled local delivery of TGF-β1 using MSs and mechanical loading is essential for neocartilage formation by MSCs and that further optimization is needed to prevent MSC differentiation towards hypertrophy.

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Section: Mscs Culture In Hydrogelmentioning

confidence: 99%

“…DNA content was measured using the QUANT-iT Picogreen, DS assay kit (Molecular Probes, Life Technologies). The total amounts of sulfated glycosaminoglycan (s-GAG) were determined by the dimethyl-methylene blue (DMMB) dye-binding assay 25 , 26 .…”

Section: Mscs Culture In Hydrogelmentioning

confidence: 99%

Cartilage tissue engineering using decellularized biomatrix hydrogel containing TGF-β-loaded alginate microspheres in mechanically loaded bioreactor

Bordbar,

Li,

Lotfibakhshaiesh

et al. 2024

Sci Rep

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“… 110 , 111 After a period of culture in vitro, methods for OA induction can be performed later on such explants. 112 , 113 The construction and shape of the model barely changed under pressure several times, so it can represent the joint in the different parts and different periods of OA severity. Grenier et al 114 constructed cartilage explants to simulate early OA injury.…”

Section: Engineered Oa Modelsmentioning

confidence: 99%

Osteoarthritis models: From animals to tissue engineering

Dou

Wang

et al. 2023

J Tissue Eng

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Osteoarthritis (OA) is a chronic degenerative osteoarthropathy. Although it has been revealed that a variety of factors can cause or aggravate the symptoms of OA, the pathogenic mechanisms of OA remain unknown. Reliable OA models that accurately reflect human OA disease are crucial for studies on the pathogenic mechanism of OA and therapeutic drug evaluation. This review first demonstrated the importance of OA models by briefly introducing the OA pathological features and the current limitations in the pathogenesis and treatment of OA. Then, it mainly discusses the development of different OA models, including animal and engineered models, highlighting their advantages and disadvantages from the perspective of pathogenesis and pathology analysis. In particular, the state-of-the-art engineered models and their potential were emphasized, as they may represent the future direction in the development of OA models. Finally, the challenges in obtaining reliable OA models are also discussed, and possible future directions are outlined to shed some light on this area.

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“…Paraffin embedded sections (4 μm thick) were mounted on glass slides for staining with Safranin O-fast green or immunohistochemistry for collagen types I and II as previously described (van Schaik et al, 2021). Briefly, paraffin cut sections were deparaffinized and treated with pepsin for 9 min at 37 °C in a humidified chamber (Digest-All 3, Thermo Fisher Scientific).…”

Section: Histology and Immunohistochemistrymentioning

confidence: 99%

Pneumatospinning Biomimetic Scaffolds for Meniscus Tissue Engineering

Dorthé

Williams²,

Grogan

et al. 2022

Front. Bioeng. Biotechnol.

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Nanofibrous scaffolds fabricated via electrospinning have been proposed for meniscus tissue regeneration. However, the electrospinning process is slow, and can only generate scaffolds of limited thickness with densely packed fibers, which limits cell distribution within the scaffold. In this study, we explored whether pneumatospinning could produce thicker collagen type I fibrous scaffolds with higher porosity, that can support cell infiltration and neo-fibrocartilage tissue formation for meniscus tissue engineering. We pneumatospun scaffolds with solutions of collagen type I with thicknesses of approximately 1 mm in 2 h. Scanning electron microscopy revealed a mix of fiber sizes with diameters ranging from 1 to 30 µm. The collagen scaffold porosity was approximately 48% with pores ranging from 7.4 to 100.7 µm. The elastic modulus of glutaraldehyde crosslinked collagen scaffolds was approximately 45 MPa, when dry, which reduced after hydration to 0.1 MPa. Mesenchymal stem cells obtained from the infrapatellar fat pad were seeded in the scaffold with high viability (>70%). Scaffolds seeded with adipose-derived stem cells and cultured for 3 weeks exhibited a fibrocartilage meniscus-like phenotype (expressing COL1A1, COL2A1 and COMP). Ex vivo implantation in healthy bovine and arthritic human meniscal explants resulted in the development of fibrocartilage-like neotissues that integrated with the host tissue with deposition of glycosaminoglycans and collagens type I and II. Our proof-of-concept study indicates that pneumatospinning is a promising approach to produce thicker biomimetic scaffolds more efficiently that electrospinning, and with a porosity that supports cell growth and neo-tissue formation using a clinically relevant cell source.

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Development of an Ex Vivo Murine Osteochondral Repair Model

Cited by 5 publications

References 48 publications

Cartilage tissue engineering using decellularized biomatrix hydrogel containing TGF-β-loaded alginate microspheres in mechanically loaded bioreactor

Cartilage tissue engineering using decellularized biomatrix hydrogel containing TGF-β-loaded alginate microspheres in mechanically loaded bioreactor

Osteoarthritis models: From animals to tissue engineering

Pneumatospinning Biomimetic Scaffolds for Meniscus Tissue Engineering

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