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Nonviral gene transfer into human meniscal cells. Part II: effect of three-dimensional environment and overexpression of human fibroblast growth factor 2

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Abstract

Purpose

Our aim was to study the effect of three-dimensional (3D) environment and overexpression of human fibroblast growth factor 2 (FGF-2) on meniscal fibrochondrocytes in vitro.

Methods

Human meniscal fibrochondrocytes were transfected with expression plasmid vectors carrying the Photinus pyralis luciferase gene, the Escherichia coli β-galactosidase gene or a human FGF-2 cDNA. Modified fibrochondrocytes were cultivated in 3D alginate hydrogel or cell pellets or in 2D monolayer culture.

Results

The levels of luciferase activity showed a peak at day two and returned to baseline levels by day 11, regardless of the type of cultivation. Both 3D environments supported the secretion of human FGF-2 protein upon FGF-2 transfection. Overexpression of human FGF-2 by genetically modified human meniscal fibrochondrocytes stimulated proliferation but not glycosaminoglycan synthesis only in 3D culture. Culture in alginate spheres resulted in a larger difference in cell numbers compared with pellet cultures.

Conclusions

Three-dimensional alginate spheres are well suited for the culture of genetically modified human meniscal fibrochondrocytes. These data are of value for cell-based approaches to meniscal repair using genetically modified human meniscal fibrochondrocytes overexpressing human FGF-2.

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Acknowledgment

We thank the members of the Department of Orthopaedic Surgery for providing human meniscal tissue.

Conflict of interest

The authors declare that they have no conflict of interest. The authors alone are responsible for the content and writing of the paper.

Authors’ contributions

HPL participated in the design of the study, carried out all studies and performed the statistical analysis. ARR participated in its design and draft. MC participated in the study design and draft. HM conceived and designed the study, co-ordinated it and drafted the manuscript. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Center of Experimental Orthopaedics, Saarland University, 66421, Homburg, Germany

    Hsiao-Ping Lee, Ana Rey-Rico, Magali Cucchiarini & Henning Madry

Authors
  1. Hsiao-Ping Lee
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  2. Ana Rey-Rico
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  3. Magali Cucchiarini
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  4. Henning Madry
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Corresponding author

Correspondence to Henning Madry.

Additional information

Presented in part at the 55th Annual Meeting of the Orthopaedic Research Society, 2009, Las Vegas, NV, USA

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Lee, HP., Rey-Rico, A., Cucchiarini, M. et al. Nonviral gene transfer into human meniscal cells. Part II: effect of three-dimensional environment and overexpression of human fibroblast growth factor 2. International Orthopaedics (SICOT) 38, 1931–1936 (2014). https://doi.org/10.1007/s00264-014-2405-z

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  • DOI: https://doi.org/10.1007/s00264-014-2405-z

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