Influence of different commercial scaffolds on the in vitro differentiation of human mesenchymal stem cells to nucleus pulposus-like cells
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- 作者：Alessandro Bertolo (1) (5)
Marco Mehr (1) (5)
Niklaus Aebli (2) (3)
Martin Baur (4)
Stephen J. Ferguson (5)
Jivko V. Stoyanov (1) (5) - 关键词：Mesenchymal stem cells ; Chondrogenesis ; Intervertebral disc ; Nucleus pulposus and three ; dimensional cultures
- 刊名：European Spine Journal
- 出版年：2012
- 出版时间：August 2012
- 年：2012
- 卷：21
- 期：6/suppl
- 页码：826-838
- 全文大小：712KB
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Marco Mehr (1) (5)
Niklaus Aebli (2) (3)
Martin Baur (4)
Stephen J. Ferguson (5)
Jivko V. Stoyanov (1) (5)
1. Swiss Paraplegic Research, 6207, Nottwil, Switzerland
5. Institute for Surgical Technology and Biomechanics, University of Bern, Stauffacherstrasse 78, Bern, Switzerland
2. Swiss Paraplegic Center, Nottwil, Switzerland
3. School of Medicine, Griffith University, Brisbane, QLD, Australia
4. Cantonal Hospital of Lucerne, Lucerne, Switzerland - ISSN：1432-0932
文摘
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.