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2 results on '"Wendy Koevoet"'

1. Differences in Cartilage-Forming Capacity of Expanded Human Chondrocytes from Ear and Nose and Their Gene Expression Profiles

Author

Catharine A. Hellingman, Eugène T. P. Verwiel, Inez Slagt, Wendy Koevoet, René M. L. Poublon, Gilbert J. Nolst-Trenité, Robert J. Baatenburg De Jong, Holger Jahr, and Gerjo J. V. M. Van Osch

Subjects
Medicine
Abstract

The aim of this study was to evaluate the potential of culture-expanded human auricular and nasoseptal chondrocytes as cell source for regeneration of stable cartilage and to analyze the differences in gene expression profile of expanded chondrocytes from these specific locations. Auricular chondrocytes in monolayer proliferated less and more slowly (two passages took 26.7 ± 2.1 days and were reached in 4.37 ± 0.30 population doublings) than nasoseptal chondrocytes (19.3 ± 2.5 days; 5.45 ± 0.20 population doublings). However, auricular chondrocytes produced larger pellets with more cartilage-like matrix than nasoseptal chondrocytes (2.2 ± 0.71 vs. 1.7 ± 0.13 mm in diameter after 35 days of culture). Although the matrix formed by auricular and nasoseptal chondrocytes contained collagen X, it did not mineralize in an in vitro model or after in vivo subcutaneous implantation. A DNA microarray study on expanded auricular and nasoseptal chondrocytes from the same donors revealed 1,090 differentially expressed genes. No difference was observed in the expression of known markers of chondrogenic capacity (e.g., collagen II, FGFR3, BMP2, and ALK1). The most striking differences were that the auricular chondrocytes had a higher expression of anabolic growth factors BMP5 and IGF1, while matrix-degrading enzymes MMP13 and ADAMTS5 were higher expressed in nasoseptal chondrocytes. This might offer a possible explanation for the observed higher matrix production by auricular chondrocytes. Moreover, chondrocytes isolated from auricular or nasoseptal cartilage had specific gene expression profiles even after expansion. These differently expressed genes were not restricted to known characterization of donor site subtype (e.g., elastic), but were also related to developmental processes.

Published
2011
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2. In Vitro Model to Study Chondrogenic Differentiation in Tendinopathy

Author

Gerjo J.V.M. van Osch, Wendy Koevoet, Nicole Kops, Hans T. M. van Schie, Jan A N Verhaar, Holger Jahr, Marieke de Mos, Orthopedics and Sports Medicine, and Otorhinolaryngology and Head and Neck Surgery

Subjects
Adult, Male, medicine.medical_specialty, Pathology, Cellular differentiation, medicine.medical_treatment, Physical Therapy, Sports Therapy and Rehabilitation, In Vitro Techniques, Achilles Tendon, Glycosaminoglycan, Young Adult, Chondrocytes, Tendinitis, SDG 3 - Good Health and Well-being, Transforming Growth Factor beta, Internal medicine, medicine, Humans, Orthopedics and Sports Medicine, Aged, Base Sequence, Models, Genetic, business.industry, Reverse Transcriptase Polymerase Chain Reaction, Growth factor, Cell Differentiation, Middle Aged, medicine.disease, Chondrogenesis, Tendon, Reverse transcription polymerase chain reaction, medicine.anatomical_structure, Endocrinology, Tendinopathy, Tissue and Organ Harvesting, Female, business
Abstract

Background Treatment of midportion Achilles tendinopathy is hampered by limited knowledge of the pathophysiology. Hypothesis Chondrogenic differentiation of tendon cells might take place in midportion Achilles tendinopathy and could be used as a target for drug treatment. An in vitro model for chondrogenic differentiation would be useful to evaluate existing and future treatment opportunities. Study Design Descriptive and controlled laboratory study. Methods Perioperatively harvested tissue from human midportion Achilles tendinotic lesions and healthy Achilles tendons was analyzed by microscopy and real-time reverse transcription polymerase chain reaction. In vitro chondrogenic differentiation of tendon explants was induced using transforming-growth-factor beta. This model was modulated by removing the chondrogenic stimulus or adding triamcinolone or platelet-rich plasma. Results Midportion Achilles tendinotic lesions had increased glycosaminoglycan staining and more rounded cell nuclei. Chondrogenic markers (sex-determining region Y)–box9, aggrecan, collagen 2, and RUNT-related transcription factor 2 were upregulated, but collagen 10 was not. Nondegenerative tendon explants cultured on chondrogenic medium had higher expression of aggrecan, collagen 2, and collagen 10 but not (sex-determining region Y)–box9 and RUNT-related transcription factor 2. Removing the chondrogenic stimulus decreased expression of aggrecan, collagen 2, and collagen 10. Both triamcinolone and platelet-rich plasma influenced the chondrogenic gene expression pattern in the in vitro model. Conclusion Chondrogenic differentiation is present in midportion Achilles tendinopathy. An in vitro model to study this chondrogenic differentiation was developed. Clinical Relevance This model can be used to investigate chondrogenic differentiation as a possible target for drug treatment, contributing to the development of more successful mechanism-based treatment opportunities.

Published
2009
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