Scaffold Free Microtissue Formation for Enhanced Cartilage Repair

Moor, Lise De; Beyls, Elien; Declercq, Heidi

doi:10.1007/s10439-019-02348-4

Cited by 51 publications

(46 citation statements)

References 56 publications

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“…However, this procedure often results in the creation of reparative tissue, with inferior quality and lacking durability when compared to the native tissue. This can be attributed to chondrocytes having the tendency to dedifferentiate during expansion in 2D culture (Dewan et al, 2014;Deng et al, 2016;Onofrillo et al, 2018;De Moor et al, 2019). Moreover, the application of individual cells fails to mimic the complex cellcell and cell-matrix interactions present within a 3D tissue, which hinders the accurate imitation of the early events in tissue development (Gionet-Gonzales and Leach, 2018).…”

Section: Discussionmentioning

confidence: 99%

“…Another commonly used approach is Matrixassisted ACI (MACI) which is an ex vivo engineered hybrid construct, where isolated autologous chondrocytes are seeded onto a biomaterial, a bovine-derived type I/III collagen scaffold, before implantation in the defect (Foldager et al, 2012;Basad et al, 2015). However, the harvesting procedure to obtain chondrocytes can induce donor-site morbidity and the 2D expansion of articular chondrocytes is characterized by long in vitro culture periods and chondrocyte dedifferentiation to a more fibroblast-like phenotype featuring a decrease in collagen II, aggrecan, and glycosaminoglycans (GAG) (Caron et al, 2012;De Moor et al, 2019). This results in the generation of repair tissue which is biochemically and hence biomechanically inferior compared to the native cartilage (Knutsen et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…Injection of spheroids can be used to treat cartilage defects instead of injecting a single cell suspension, whose dedifferentiation leads to inferior fibrous tissue. Spheroid maturation already starts in vitro, establishing a tissue specific ECM provoked by their high cellularity and cell density (Huang et al, 2016;De Moor et al, 2019). An example used in clinical trials are chondrospheres, 500-800 µm spheroids generated from autologous chondrocytes.…”

Section: Introductionmentioning

confidence: 99%

“…Though for bioprinting, small diameter spheroids with uniform shapes and sizes are necessary for compatibility with the print needle. In our previous study, we described the biofabrication of printable high quality cartilage microtissues starting from porcine chondrocytes (De Moor et al, 2019). However, to enhance clinical translation, we describe the high-throughput creation of small diameter human cartilage spheroids (<200 µm) by chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSC), applying chondrogenic culture medium and low oxygen tension (5%).…”

Section: Introductionmentioning

confidence: 99%

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Hybrid Bioprinting of Chondrogenically Induced Human Mesenchymal Stem Cell Spheroids

Moor

Fernandez

Vercruysse

et al. 2020

Front. Bioeng. Biotechnol.

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To date, the treatment of articular cartilage lesions remains challenging. A promising strategy for the development of new regenerative therapies is hybrid bioprinting, combining the principles of developmental biology, biomaterial science, and 3D bioprinting. In this approach, scaffold-free cartilage microtissues with small diameters are used as building blocks, combined with a photo-crosslinkable hydrogel and subsequently bioprinted. Spheroids of human bone marrow-derived mesenchymal stem cells (hBM-MSC) are created using a high-throughput microwell system and chondrogenic differentiation is induced during 42 days by applying chondrogenic culture medium and low oxygen tension (5%). Stable and homogeneous cartilage spheroids with a mean diameter of 116 ± 2.80 µm, which is compatible with bioprinting, were created after 14 days of culture and a glycosaminoglycans (GAG)and collagen II-positive extracellular matrix (ECM) was observed. Spheroids were able to assemble at random into a macrotissue, driven by developmental biology tissue fusion processes, and after 72 h of culture, a compact macrotissue was formed. In a directed assembly approach, spheroids were assembled with high spatial control using the bio-ink based extrusion bioprinting approach. Therefore, 14-day spheroids were combined with a photo-crosslinkable methacrylamide-modified gelatin (gelMA) as viscous printing medium to ensure shape fidelity of the printed construct. The photo-initiators Irgacure 2959 and Li-TPO-L were evaluated by assessing their effect on bio-ink properties and the chondrogenic phenotype. The encapsulation in gelMA resulted in further chondrogenic maturation observed by an increased production of GAG and a reduction of collagen I. Moreover, the use of Li-TPO-L lead to constructs with lower stiffness which induced a decrease of collagen I and an increase in GAG and collagen II production. After 3D bioprinting, spheroids remained viable and the cartilage phenotype was maintained. Our findings demonstrate that hBM-MSC spheroids are able to differentiate into cartilage microtissues and display a geometry compatible with 3D bioprinting. Furthermore, for hybrid bioprinting of these spheroids,

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Hybrid Bioprinting of Chondrogenically Induced Human Mesenchymal Stem Cell Spheroids

Moor

Fernandez

Vercruysse

et al. 2020

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

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“…Musculoskeletal research of the joint classically relies on distinct in-vitro and in-vivo approaches. In vitro models are based on cell lines (6) monolayer cultures (7) and cultures providing a three-dimensional (3D) cell surrounding (8) (9). While being the most economic and ethically supportable choice for investigation, all these models do not su ciently represent the native situation.…”

mentioning

confidence: 99%

Osteochondral Live Slices as Physiological in-vitro Model of the Human Joint for High-throughput Applications

Spinnen

Rendenbach

Kühl

et al. 2020

Preprint

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Background: Current advances in musculoskeletal research yielded numerous new insights and therapy options for diseases of the joints but preclinical in-vitro testing is currently limited to self-manufactured 2D and 3D cell cultures only remotely mimicking the in-vivo situation in regards of tissue composition and cell configuration. While in-vivo animal models are a lot more lifelike, they are inherently connected with ethical concerns, translation to the human situation, logistic efforts and high costs. Here we explored the use of native human life slice cultures from explanted tibial plateaus upon their feasibility to serve as a highly lifelike and easy-to-handle model of the human joint which can be obtained in masses from few tissue samples.Methods: Osteochondral slices (1cm x 1cm x 500µm) were prepared with a special microtome from 23 human tibial plateaus and subsequently cultivated in hanging inserts over the course of 7 or 21 days. Short-term cultivated slices were stimulated with either 800 pM/1.2 nM TNF-α or 800 pM TGF-β3. During cultivation, viability of tissue cells was assessed via laser microscopy / resazurin assay. After cultivation, gene expression of cartilage ECM Marker proteins was quantified with RT-PCR. TNF-α stimulated slices and their controls were stained with Safranin-O and analyzed via histomorphometry to quantify tissue proteoglycan content. Results: Tissue cell viability remained >90% over the three weeks. Laser scanning microscopy revealed highly conserved spatial alignment of cells inside the cartilage. Incubation of the slice cultures with TNF-α showed a significant, dose dependent decrease in mRNA expression of cartilage proteins collagen II, cartilage oligomeric matrix protein and aggrecan while incubation with TGF-β3 caused a significant increase of early bone formation proteins collagen I and also cartilage oligomeric matrix protein. Histologically, TNF-α incubation caused a 32% reduction of proteoglycans, detachment of collagen fibers and cell swelling.Conclusion: In summary, native osteochondral slice cultures provide a stable and manipulable, physiological model of human joint bone and cartilage biology. The setup is easy to produce and handle, scalable, and contributes to 3R principles in biomedical research, suggesting itself as a platform for preclinical testing especially regarding high-throughput applications.

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Hyaluronic Acid‐based Cell‐free Composite Scaffold Promotes Osteochondral Repair In Vitro by Upregulating Osteogenic‐ and Chondrogenic‐Specific Gene Expressions

Pathmanapan,

Muthuramalingam,

Pandurangan

et al. 2024

Adv Eng Mater

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Osteochondral defects pose a significant challenge in clinical practice, leading to increased medical care, diminished quality of life for patients, and elevated economic burden. The restoration of osteochondral defects, particularly in cartilage, is limited by its finite repair capacity and complex architecture. Treatment based on regenerative therapies acclaims the favorable alternative to contemporary procedures. Studies reveal that engineered biomaterials hold the significant importance in stimulating tissue repair with minimal cost and risks. In this study, we developed a porous composite scaffold, where host cells infiltrate and creates the microenvironment for cellular adhesion and enhanced differentiation. Thus, we fabricated a composite scaffold composed of natural glycosaminoglycan (GAG) hyaluronic acid (HA), the protein component fibrin, bio‐ceramic nano hydroxy apatite (nHAP) and graphene oxide (GO). Scanning electron microscopy (SEM) observation and physiochemical characterization revealed an interconnected pore structure, optimum swelling potential, high porosity (80.14%), controlled biodegradation, high mechanical properties and mineralization. Evaluation of scaffold’s biocompatibility using osteoblast ‐ like MG‐63 cells, showed adequate viability, cell adhesion, osteoinductive potential. The upregulated expressions of osteogenic and chondrogenic genes OCN, ALP, COL1A1, ACAN, SOX9 and COL2A1 promotes mineralization and ECM formation. These findings suggest that the composite scaffold HA‐GO‐F‐nHAP exhibits promising potential for osteochondral repair.This article is protected by copyright. All rights reserved.

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Scaffold Free Microtissue Formation for Enhanced Cartilage Repair

Cited by 51 publications

References 56 publications

Hybrid Bioprinting of Chondrogenically Induced Human Mesenchymal Stem Cell Spheroids

Hybrid Bioprinting of Chondrogenically Induced Human Mesenchymal Stem Cell Spheroids

Osteochondral Live Slices as Physiological in-vitro Model of the Human Joint for High-throughput Applications

Hyaluronic Acid‐based Cell‐free Composite Scaffold Promotes Osteochondral Repair In Vitro by Upregulating Osteogenic‐ and Chondrogenic‐Specific Gene Expressions

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