Optimization of Chondrocyte Isolation and Phenotype Characterization for Cartilage Tissue Engineering

Lau, Ting Ting; Peck, Yvonne; Huang, Weiliang; Wang, Dong‐An

doi:10.1089/ten.tec.2014.0159

Cited by 20 publications

(12 citation statements)

References 22 publications

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“…Female articular chondrocytes (AC) were isolated from healthy donors as per published method 26, 27 . The AC cells were cultured in 24 well plates and treated with OA and NON-OA synovial fluid derived EVs (40 µg/ml) separately with 1% FBS (exosome free) media for 48 hrs.…”

Section: Methodsmentioning

confidence: 99%

Gender-specific differential expression of exosomal miRNA in synovial fluid of patients with osteoarthritis

Kolhe

Hunter

Liu

et al. 2017

Sci Rep

174

171

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The pathogenesis of osteoarthritis (OA) is poorly understood, and therapeutic approaches are limited to preventing progression of the disease. Recent studies have shown that exosomes play a vital role in cell-to-cell communication, and pathogenesis of many age-related diseases. Molecular profiling of synovial fluid derived exosomal miRNAs may increase our understanding of OA progression and may lead to the discovery of novel biomarkers and therapeutic targets. In this article we report the first characterization of exosomes miRNAs from human synovial fluid. The synovial fluid exosomes share similar characteristics (size, surface marker, miRNA content) with previously described exosomes in other body fluids. MiRNA microarray analysis showed OA specific exosomal miRNA of male and female OA. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis identified gender-specific target genes/signaling pathways. These pathway analyses showed that female OA specific miRNAs are estrogen responsive and target TLR (toll-like receptor) signaling pathways. Furthermore, articular chondrocytes treated with OA derived extracellular vesicles had decreased expression of anabolic genes and elevated expression of catabolic and inflammatory genes. In conclusion, synovial fluid exosomal miRNA content is altered in patients with OA and these changes are gender specific.

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Section: Methodsmentioning

confidence: 99%

Gender-specific differential expression of exosomal miRNA in synovial fluid of patients with osteoarthritis

Kolhe

Hunter

Liu

et al. 2017

Sci Rep

174

171

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“…Comparing our data to this report, suggests that the cartilage assessed includes both non‐degenerate and degenerate cartilage. It should be noted that tissue isolation, processing, and enzymatic digestion protocols play a significant role in the outcomes . [Cells] isolated from cartilage reported in literature vary by 30‐fold, from as low as 0.17 million cells/gram up to 5 million cells/gram .…”

Section: Discussionmentioning

confidence: 99%

Progenitor cells from different zones of human cartilage and their correlation with histopathological osteoarthritis progression

Mantripragada

Bova

Boehm

et al. 2017

Journal Orthopaedic Research

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Cell-based therapies development for the treatment of osteoarthritis (OA) requires an understanding of the disease progression and attributes of the cells resident in cartilage. This study focused on quantitative assessment of the concentration and biological potential of stem and progenitor cells resident in different zones of cartilage displaying macroscopic Outerbridge grade 1-2 OA, and their correlation with OA progression based on established histologic scoring system. Lateral femoral condyles were collected from 15 patients with idiopathic OA and varus knees undergoing total knee arthroplasty. Superficial(C , top ∼ 500 µm) and deep cartilage(C ) was separated. Chondrogenic Connective Tissue Progenitors (CTP-C) were assayed by standardized Colony-Forming-Unit assay using automated image analysis (Colonyze ) based on ASTM standard F-2944-12. Cell concentration (cells/mg) was significantly greater in C (median: 7,000; range: 3,440-17,600) than C (median: 5,340; range: 3,393-9,660), p = 0.039. Prevalence (CTPs/million cells) was not different between C (median: 1,274; range: 0-3,898) and C (median:1,365; range:0-6,330), p = 0.42. In vitro performance of CTP-C progeny varied widely within and between patients, manifest by variation in colony size and morphology. Mean histopathological Mankin score was 4.7 (SD = 1.2), representing mild to moderate OA. Tidemark breach by blood vessels was associated with lower C cell concentration (p = 0.02). Matrix degradation was associated with lower C cell and CTP-C concentration (p = 0.015 and p = 0.095, respectively), independent of articular surface changes. These findings suggest that the initiation of OA may occur in either superficial or deep zones. The pathological changes affect CTP-Cs in C and C cartilage zones differently. The heterogeneity among the available CTP-Cs in C and C suggests performance-based selection to optimize cell-sourcing strategies for therapy. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1728-1738, 2018.

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“…Various methods have been utilized in order to achieve maximum harvest yield with optimal cell viability while preserving the chondrocyte phenotype [11][12][13]. Majority of these are implemented on animal chondrocytes that are usually used as control cells in the experiments with engineered constructs.…”

Section: Cells As Biomimetic System Componentmentioning

confidence: 99%

Mimetic Hierarchical Approaches for Osteochondral Tissue Engineering

Gadjanski

2018

Osteochondral Tissue Engineering

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In order to engineer biomimetic osteochondral (OC) construct, it is necessary to address both the cartilage and bone phase of the construct, as well as the interface between them, in effect mimicking the developmental processes when generating hierarchical scaffolds that show gradual changes of physical and mechanical properties, ideally complemented with the biochemical gradients. There are several components whose characteristics need to be taken into account in such biomimetic approach, including cells, scaffolds, bioreactors as well as various developmental processes such as mesenchymal condensation and vascularization, that need to be stimulated through the use of growth factors, mechanical stimulation, purinergic signaling, low oxygen conditioning, and immunomodulation. This chapter gives overview of these biomimetic OC system components, including the OC interface, as well as various methods of fabrication utilized in OC biomimetic tissue engineering (TE) of gradient scaffolds. Special attention is given to addressing the issue of achieving clinical size, anatomically shaped constructs. Besides such neotissue engineering for potential clinical use, other applications of biomimetic OC TE including formation of the OC tissues to be used as high-fidelity disease/ healing models and as in vitro models for drug toxicity/efficacy evaluation are covered. Highlights Biomimetic OC TE uses "smart" scaffolds able to locally regulate cell phenotypes and dual-flow bioreactors for two sets of conditions for cartilage/ bone Protocols for hierarchical OC grafts engineering should entail mesenchymal condensation for cartilage and vascular component for bone Immunomodulation, low oxygen tension, purinergic signaling, time dependence of stimuli application are important aspects to consider in biomimetic OC TE Mimetic Hierarchical Approaches for Osteochondral Tissue Engineering Ivana GadjanskiAbstract In order to engineer biomimetic osteochondral (OC) construct, it is necessary to address both the cartilage and bone phase of the construct, as well as the interface between them, in effect mimicking the developmental processes when generating hierarchical scaffolds that show gradual changes of physical and mechanical properties, ideally complemented with the biochemical gradients. There are several components whose characteristics need to be taken into account in such biomimetic approach, including cells, scaffolds, bioreactors as well as various developmental processes such as mesenchymal condensation and vascularization, that need to be stimulated through the use of growth factors, mechanical stimulation, purinergic signaling, low oxygen conditioning, and immunomodulation. This chapter gives overview of these biomimetic OC system components, including the OC interface, as well as various methods of fabrication utilized in OC biomimetic tissue engineering (TE) of gradient scaffolds. Special attention is given to addressing the issue of achieving clinical size, anatomically shaped constructs. Besides such neotissue en...

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Optimization of Chondrocyte Isolation and Phenotype Characterization for Cartilage Tissue Engineering

Cited by 20 publications

References 22 publications

Gender-specific differential expression of exosomal miRNA in synovial fluid of patients with osteoarthritis

Gender-specific differential expression of exosomal miRNA in synovial fluid of patients with osteoarthritis

Progenitor cells from different zones of human cartilage and their correlation with histopathological osteoarthritis progression

Mimetic Hierarchical Approaches for Osteochondral Tissue Engineering

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