Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in vitro redifferentiation.
Abstract: If chondrocytes from the superficial, middle, and deep zones of articular cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. Objective: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture). Methods: Controlled laboratory study. Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes). Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone. Conclusions: Chondrocytes isolated from the 3 zones of equine cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion. Conclusions: Appreciation of the zonal differences can lead to important advances in cartilage tissue engineering. Findings support the use of hydrogels such as alginate for engineering zonal cartilage constructs.
Publication Date: 2009-10-21 PubMed ID: 19846691DOI: 10.1177/0363546509350978Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study investigates the possibility of maintaining or regaining specific characteristics of chondrocytes, cells found in different zones of articular cartilage, during in vitro culture. The research shows that these cells lose their zone-specific traits during 2D expansion but can reacquire them in a 3D environment. Specifically, culturing chondrocytes in a material called alginate allows them to regain their zone-specific qualities, which can enhance cartilage tissue engineering approaches.
Research Methodology
- The researchers used controlled laboratory study methods for this investigation.
- Articular chondrocytes from three different zones (superficial, middle, and deep) of equine cartilage were expanded using a method known as monolayer culture.
- After the monolayer culture phase, the chondrocytes underwent redifferentiation in two types of cultures: pellet and alginate bead, for a period of up to four weeks.
- The researchers examined various aspects of these cells, including quantifying the levels of glycosaminoglycans and DNA, and assessing the expression of different zonal markers through immunohistochemical testing. These markers included the cartilage oligomeric protein, indicative of cells from deeper zones, and clusterin, expressed specifically by superficial chondrocytes.
Results and Findings
- There was a variation in the yield of cells obtained from different cartilage zones, but the rates of cell proliferation didn’t differ significantly.
- All of the zonal markers under evaluation were lost when the chondrocytes underwent expansion.
- Following redifferentiation, the cells cultured in pellet forms produced more glycosaminoglycans per DNA amount compared to those cultured in alginate beads.
- However, unlike cells in pellet cultures, the chondrocytes in alginate beads were able to regain zonal differences characterized by the reappearance of zone-specific markers, such as the cartilage oligomeric protein and clusterin. In addition, cells from the deep zone produced significantly more glycosaminoglycans compared to those from the superficial zone.
Conclusions and Implications
- The findings suggest that when cultured in vitro in alginate after expansion, chondrocytes sourced from the three zones of equine cartilage have the ability to restore their zonal matrix expression.
- This appreciation of zonal differences can potentially drive significant advancements in the field of cartilage tissue engineering, leading to better and more efficient techniques.
- Overall, the study recommends the use of hydrogels like alginate to engineer zonal cartilage constructs due to their ability to promote redifferentiation and recreate zonal differences in cartilage cells.
Cite This Article
APA
Schuurman W, Gawlitta D, Klein TJ, ten Hoope W, van Rijen MH, Dhert WJ, van Weeren PR, Malda J.
(2009).
Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in vitro redifferentiation.
Am J Sports Med, 37 Suppl 1, 97S-104S.
https://doi.org/10.1177/0363546509350978 Publication
Researcher Affiliations
- Department of Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands.
MeSH Terms
- Animals
- Biomarkers
- Cartilage / growth & development
- Cartilage, Articular / cytology
- Cell Culture Techniques / methods
- Cell Differentiation / physiology
- Cell Proliferation
- Chondrocytes / cytology
- Chondrocytes / metabolism
- Chondrogenesis / physiology
- Extracellular Matrix / genetics
- Extracellular Matrix / metabolism
- Glycosaminoglycans / genetics
- Glycosaminoglycans / metabolism
- Horses
- Immunohistochemistry
- Tissue Engineering / methods
Citations
This article has been cited 18 times.- Liu W, Madry H, Cucchiarini M. Application of Alginate Hydrogels for Next-Generation Articular Cartilage Regeneration.. Int J Mol Sci 2022 Jan 20;23(3).
- Naghieh S, Lindberg G, Tamaddon M, Liu C. Biofabrication Strategies for Musculoskeletal Disorders: Evolution towards Clinical Applications.. Bioengineering (Basel) 2021 Sep 10;8(9).
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- Peiffer QC, de Ruijter M, van Duijn J, Crottet D, Dominic E, Malda J, Castilho M. Melt electrowriting onto anatomically relevant biodegradable substrates: Resurfacing a diarthrodial joint.. Mater Des 2020 Oct;195:109025.
- Owida HA, Yang R, Cen L, Kuiper NJ, Yang Y. Induction of zonal-specific cellular morphology and matrix synthesis for biomimetic cartilage regeneration using hybrid scaffolds.. J R Soc Interface 2018 Jun;15(143).
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