☆ 4.4 Article

High-throughput bone and cartilage micropellet manufacture, followed by assembly of micropellets into biphasic osteochondral tissue

CELL AND TISSUE RESEARCH (2015)

期刊

CELL AND TISSUE RESEARCH

卷 361, 期 3, 页码 755-768

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SPRINGER

DOI: 10.1007/s00441-015-2159-y

关键词

Articular cartilage; Tissue engineering; Osteochondral defect repair; Mesenchymal stem/stromal cells; Micropellets; Induction media; Human

类别

Cell Biology

资金

National Health and Medical Research Council of Australia (NHMRC)

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摘要

Engineered biphasic osteochondral tissues may have utility in cartilage defect repair. As bone-marrow-derived mesenchymal stem/stromal cells (MSC) have the capacity to make both bone-like and cartilage-like tissues, they are an ideal cell population for use in the manufacture of osteochondral tissues. Effective differentiation of MSC to bone-like and cartilage-like tissues requires two unique medium formulations and this presents a challenge both in achieving initial MSC differentiation and in maintaining tissue stability when the unified osteochondral tissue is subsequently cultured in a single medium formulation. In this proof-of-principle study, we used an in-house fabricated microwell platform to manufacture thousands of micropellets formed from 166 MSC each. We then characterized the development of bone-like and cartilage-like tissue formation in the micropellets maintained for 8-14 days in sequential combinations of osteogenic or chondrogenic induction medium. When bone-like or cartilage-like micropellets were induced for only 8 days, they displayed significant phenotypic changes when the osteogenic or chondrogenic induction medium, respectively, was swapped. Based on these data, we developed an extended 14-day protocol for the pre-culture of bone-like and cartilage-like micropellets in their respective induction medium. Unified osteochondral tissues were formed by layering 12,000 osteogenic micropellets and 12,000 chondrogenic micropellets into a biphasic structure and then further culture in chondrogenic induction medium. The assembled tissue was cultured for a further 8 days and characterized via histology. The micropellets had amalgamated into a continuous structure with distinctive bone-like and cartilage-like regions. This proof-of-concept study demonstrates the feasibility of micropellet assembly for the formation of osteochondral-like tissues for possible use in osteochondral defect repair.

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Comparison between articular chondrocytes and mesenchymal stromal cells for the production of articular cartilage implants

Nadine Frerker, Tommy A. A. Karlsen, Maria Stensland, Tuula A. A. Nyman, Simon Rayner, Jan E. E. Brinchmann

Summary: Focal lesions of articular cartilage can cause pain and reduced joint function, leading to osteoarthritis if untreated. In vitro generated, scaffold-free autologous cartilage discs made from articular chondrocytes were compared to those made from bone marrow-derived mesenchymal stromal cells. Articular chondrocytes produced more extracellular matrix and contained more articular cartilage proteins, while mesenchymal stromal cells had more proteins associated with cartilage hypertrophy and bone formation. MicroRNA analysis suggested that differential expression of microRNAs played a key role in the differential synthesis of proteins. Therefore, articular chondrocytes are preferred for engineering articular cartilage.

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