Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage - PubMed (original) (raw)

Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage

Liming Bian et al. Tissue Eng Part A. 2011 Apr.

Abstract

Mesenchymal stem cells (MSCs) are being recognized as a viable cell source for cartilage repair; however, it still remains a challenge to recapitulate the functional properties of native articular cartilage using only MSCs. Additionally, MSCs may exhibit a hypertrophic phenotype under chondrogenic induction, resulting in calcification after ectopic transplantation. With this in mind, the objective of this study was to assess whether the addition of chondrocytes to MSC cultures influences the properties of tissue-engineered cartilage and MSC hypertrophy when cultured in hyaluronic acid hydrogels. Mixed cell populations (human MSCs and human chondrocytes at a ratio of 4:1) were encapsulated in the hydrogels and exhibited significantly higher Young's moduli, dynamic moduli, glycosaminoglycan levels, and collagen content than did constructs seeded with only MSCs or chondrocytes. Furthermore, the deposition of collagen X, a marker of MSC hypertrophy, was significantly lower in the coculture constructs than in the constructs seeded with MSCs alone. When MSCs and chondrocytes were cultured in distinct gels, but in the same wells, there was no improvement in biomechanical and biochemical properties of the engineered tissue, implying that a close proximity is essential. This approach can be used to improve the properties and prevent calcification of engineered cartilage formed from MSC-seeded hydrogels with the addition of lower fractions of chondrocytes, leading to improved clinical outcomes.

PubMed Disclaimer

Figures

FIG. 1.

FIG. 1.

Cell types (human MSCs and chondrocytes) and seeding densities of experimental groups. MSCs, mesenchymal stem cells.

FIG. 2.

FIG. 2.

Viability staining of cells in HA hydrogels on days 14 and 35. Green: live cells; red: dead cells; scale bar = 100 μm. HA, hyaluronic acid. Color images available online at

www.liebertonline.com/tea

.

FIG. 3.

FIG. 3.

Peak stress (A), dynamic modulus (1 Hz) (B), Young's modulus (C), GAG (D), total collagen (E), and DNA (F) content of all experimental groups (data presented as mean ± standard deviation). ND: not detectable, *p < 0.05 versus all other groups at the same culture time; †p < 0.05 versus MSC at the same culture time (n = 4). The Chon20 group was not tested for mechanical properties due to poor early cell viability. GAG, glycosaminoglycan.

FIG. 4.

FIG. 4.

Gene expression (in fold change) of selected chondrogenic and hypertrophic markers with time in culture (data presented as mean ± standard deviation). Only the Mixed and MSC groups were compared due to limited viability in other experimental groups. *p < 0.05 versus MSC group (n = 3).

FIG. 5.

FIG. 5.

Immunohistochemical staining for chondroitin sulfate (CS), type II collagen, and type I collagen on day 42; bar in inset = 50 μm. Color images available online at

www.liebertonline.com/tea

.

FIG. 6.

FIG. 6.

Immunohistochemical staining for collagen 10A1 on day 42; bar in inset = 25 μm. Color images available online at

www.liebertonline.com/tea

.

FIG. 7.

FIG. 7.

Young's modulus (A), DNA (B), GAG (C), and total collagen (D) content of all groups on day 42. The entities 19:1, 16:4, 10:10, and 20:0 represent ratios of MSCs and chondrocytes seeded in mixture in terms of million cells/mL (i.e., 19:1 indicates a seeding density of 19 million MSCs and 1 million chondrocytes per mL of hydrogel and cell mixture, data presented as mean ± standard deviation). *p < 0.05 versus 19:1 and 16:4 group (n = 4).

Similar articles

Cited by

References

    1. Williams C.G. Kim T.K. Taboas A. Malik A. Manson P. Elisseeff J. In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel. Tissue Eng. 2003;9:679. - PubMed
    1. Erickson I.E. Huang A.H. Chung C. Li R.T. Burdick J.A. Mauck R.L. Differential maturation and structure-function relationships in mesenchymal stem cell- and chondrocyte-seeded hydrogels. Tissue Eng Part A. 2009;15:1041. - PMC - PubMed
    1. Friedenstein A.J. Piatetzky S., II Petrakova K.V. Osteogenesis in transplants of bone marrow cells. J Embryol Exp Morphol. 1966;16:381. - PubMed
    1. Pittenger M.F. Mackay A.M. Beck S.C. Jaiswal R.K. Douglas R. Mosca J.D., et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143. - PubMed
    1. Chung C. Beecham M. Mauck R.L. Burdick J.A. The influence of degradation characteristics of hyaluronic acid hydrogels on in vitro neocartilage formation by mesenchymal stem cells. Biomaterials. 2009;30:4287. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources