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Role of polyamines in hypertrophy and terminal differentiation of osteoarthritic chondrocytes

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Abstract

Polyamines are naturally occurring, positively charged polycations which are able to control several cellular processes in different cell types, by interacting with negatively charged compounds and structures within the living cell. Functional genomics in rodents targeting key biosynthetic or catabolic enzymes have revealed a series of phenotypic changes, many of them related to human diseases. Several pieces of evidence from the literature point at a role of polyamines in promoting chondrocyte differentiation, a process which is physiological in growth plate maturation or fracture healing, but has pathological consequences in articular chondrocytes, programmed to keep a maturational arrested state. Inappropriate differentiation of articular chondrocytes results in osteoarthritis. Thus, we have studied the effects of exogenously added spermine or spermidine in chondrocyte maturation recapitulated in 3D cultures, to tease out the effects on gene and protein expression of key chondrogenesis regulatory transcription factors, markers and effectors, as well as their posttranscriptional regulation. The results indicate that both polyamines are able to increase the rate and the extent of chondrogenesis, with enhanced collagen 2 deposition and remodeling with downstream generation of collagen 2 bioactive peptides. These were able to promote nuclear localization of RUNX-2, the pivotal transcription factor in chondrocyte hypertrophy and osteoblast generation. Indeed, samples stimulated with polyamines showed an enhanced mineralization, along with increased caspase activity, indicating increased chondrocyte terminal differentiation. In conclusion these results indicate that the polyamine pathway can represent a potential target to control and correct chondrocyte inappropriate maturation in osteoarthritis.

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Abbreviations

AR-S:

Alizarin red-S

DDR:

Discoidin domain receptors

DFMO:

α-Difluoromethylornithine

ECM:

Extra-cellular matrix

FBS:

Fetal bovine serum

GAG:

Glycosaminoglycans

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

KD:

Knockdown

MMP:

Matrix metalloproteinase

OA:

Osteoarthritis

ODC:

Ornithine decarboxylase

PCR:

Polymerase chain reaction

PTH:

Parathormone

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Acknowledgments

This work was supported by grants from Università di Bologna (R.F.O.), Istituto Ortopedico Rizzoli, MIUR (PRIN and FIRB), Carisbo Foundation and Fondi cinque per mille, Ministero della Salute, Italy. We acknowledge the valuable contribution of Prof. Kenneth B. Marcu, Stony Brook University, USA, to support us in the retroviral delivery of IKKα shRNA to primary chondrocytes.

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Authors and Affiliations

  1. Dipartimento di Biochimica “G. Moruzzi”, University of Bologna, Bologna, Italy

    A. Facchini, S. Cetrullo & F. Flamigni

  2. Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, Bologna, Italy

    R. M. Borzì, E. Olivotto, D. Platano & S. Pagani

  3. Dipartimento RIT, Laboratorio RAMSES, Istituto Ortopedico Rizzoli, Bologna, Italy

    R. M. Borzì & E. Olivotto

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  1. A. Facchini
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  2. R. M. Borzì
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  3. E. Olivotto
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  4. D. Platano
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  5. S. Pagani
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  6. S. Cetrullo
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  7. F. Flamigni
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Correspondence to F. Flamigni.

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Facchini, A., Borzì, R.M., Olivotto, E. et al. Role of polyamines in hypertrophy and terminal differentiation of osteoarthritic chondrocytes. Amino Acids 42, 667–678 (2012). https://doi.org/10.1007/s00726-011-1041-9

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  • DOI: https://doi.org/10.1007/s00726-011-1041-9

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