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Influence of different commercial scaffolds on the in vitro differentiation of human mesenchymal stem cells to nucleus pulposus-like cells

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Abstract

Introduction

Cell-based therapies for regeneration of the degenerated intervertebral disc (IVD) are an alternative to current surgical intervention. Mesenchymal stem cells (MSCs), in combination with a scaffold, might be ideal candidates for regenerating nucleus pulposus (NP), the pressure-distributing part of the IVD. While the use of growth factors for MSCs differentiation currently receives major attention, in this study we compare the performance of sponge-like matrixes in supporting cell differentiation into NP-like cells.

Materials and methods

Four types matrixes approved as medical devices for other applications were tested as scaffolds for MSCs: two made of equine or porcine collagen, one of gelatin and one of chitosan. Bone marrow-derived human MSCs were seeded in these scaffolds or embedded in alginate, as a three-dimensional control. After five weeks in culture, NP-like differentiation of the cell-scaffold constructs was analyzed by qRT-PCR, histology, total DNA quantification, proteoglycan accumulation and immunohistochemistry.

Results

MSCs in collagen matrixes and gelatin produced more mRNA and proteins of the chondrogenic markers collagen type I, collagen type II (COL2) and aggrecan (ACAN), when compared with cells embedded in alginate or chitosan. Proteoglycan accumulation and cell survival were also higher in collagen and gelatin matrixes. Gene expression results were also confirmed by histological and immunohistochemical staining. In contrast to alginate control, the gene expression of the undesired bone marker osteopontin was lower in all tested groups. In porcine collagen supports, MSC expression ratio between COL2/ACAN closely resembled the expression of nucleus pulposus cells, but gene expression of recently described NP markers keratin19, PAX1 and FOXF1 was lower.

Conclusions

Collagen supports provide a readily available, medically approved and effective scaffold for chondrogenic differentiation in vitro, but the phenotype of differentiated MSCs is not yet completely equivalent to that of NP cells.

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Acknowledgments

This work was supported by the Swiss Paraplegic Foundation. We thank Mr. Beat Haenni, from the Institute of Anatomy of Bern, for technical assistance with scanning electron microscopy.

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Authors and Affiliations

  1. Swiss Paraplegic Research, 6207, Nottwil, Switzerland

    Alessandro Bertolo, Marco Mehr & Jivko V. Stoyanov

  2. Swiss Paraplegic Center, Nottwil, Switzerland

    Niklaus Aebli

  3. School of Medicine, Griffith University, Brisbane, QLD, Australia

    Niklaus Aebli

  4. Cantonal Hospital of Lucerne, Lucerne, Switzerland

    Martin Baur

  5. Institute for Surgical Technology and Biomechanics, University of Bern, Stauffacherstrasse 78, Bern, Switzerland

    Alessandro Bertolo, Marco Mehr, Stephen J. Ferguson & Jivko V. Stoyanov

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  1. Alessandro Bertolo
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Correspondence to Jivko V. Stoyanov.

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Bertolo, A., Mehr, M., Aebli, N. et al. Influence of different commercial scaffolds on the in vitro differentiation of human mesenchymal stem cells to nucleus pulposus-like cells. Eur Spine J 21 (Suppl 6), 826–838 (2012). https://doi.org/10.1007/s00586-011-1975-3

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  • DOI: https://doi.org/10.1007/s00586-011-1975-3

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