Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration

F.C. Bach, S.A.H. de Vries, F.M. Riemers, J. Boere, F.W.M. van Heel, M. van Doeselaar, S.S. Goerdayal, P.G.J. Nikkels, K. Benz, L. B. Creemers, A.F.M. Altelaar, B.P. Meij, K. Ito, M.A. Tryfonidou

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Abstract

During intervertebral disc (IVD) maturation, notochordal cells (NCs) are replaced by chondrocyte-like cells (CLCs) in the nucleus pulposus, suggesting that NCs play a role in maintaining tissue health. Affirmatively, NC-conditioned medium (NCCM) exerts regenerative effects on CLC proliferation and extracellular matrix (ECM) production. The aim of this study was to identify NC-secreted substances that stimulate IVD regeneration. By mass spectrometry of porcine, canine and human NCCM, 149, 170 and 217 proteins were identified, respectively, with 66 proteins in common. Mainly ECM-related proteins were identified, but also organelle-derived and membrane-bound vesicle proteins. To determine whether the effect of NCCM was mediated by soluble and/or pelletable factors, porcine and canine NCCM were separated into a soluble (NCCM-S; peptides and proteins) and pelletable (NCCM-P; protein aggregates and extracellular vesicles) fraction by ultracentrifugation, and tested on bovine and canine CLCs in vitro, respectively. In each model, NCCM-S exerted a more pronounced anabolic effect than NCCM-P. However, glycosaminoglycan (GAG) uptake from the medium into the carrier gel prevented more definite conclusions. While the effect of porcine NCCM-P on bovine CLCs was negligible, canine NCCM-P appeared to enhance GAG and collagen type II deposition by canine CLCs. In conclusion, porcine and canine NCCM exerted their anabolic effects mainly through soluble factors, but also the pelletable NCCM factors showed moderate regenerative potential. Although the regenerative potential of NCCM-P should not be overlooked, future studies should focus on unraveling the protein-based regenerative mechanism from NCCM produced from isolated NCs, e.g. by NCCM fractionation and pathway blocking studies.
Original languageEnglish
Pages (from-to)163-180
JournalEuropean Cells and Materials
Volume32
DOIs
Publication statusPublished - 2016

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Intervertebral Disc
Conditioned Culture Medium
Canidae
Regeneration
Swine
Proteins
Chondrocytes
Anabolic Agents
Cell proliferation
Fractionation
Collagen
Glycosaminoglycans
Peptides
Mass spectrometry
Gels
Health
Tissue
Membranes
Collagen Type II
Extracellular Matrix Proteins

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Bach, F. C., de Vries, S. A. H., Riemers, F. M., Boere, J., van Heel, F. W. M., van Doeselaar, M., ... Tryfonidou, M. A. (2016). Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration. European Cells and Materials, 32, 163-180. https://doi.org/10.22203/eCM.v032a11
Bach, F.C. ; de Vries, S.A.H. ; Riemers, F.M. ; Boere, J. ; van Heel, F.W.M. ; van Doeselaar, M. ; Goerdayal, S.S. ; Nikkels, P.G.J. ; Benz, K. ; Creemers, L. B. ; Altelaar, A.F.M. ; Meij, B.P. ; Ito, K. ; Tryfonidou, M.A. / Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration. In: European Cells and Materials. 2016 ; Vol. 32. pp. 163-180.
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abstract = "During intervertebral disc (IVD) maturation, notochordal cells (NCs) are replaced by chondrocyte-like cells (CLCs) in the nucleus pulposus, suggesting that NCs play a role in maintaining tissue health. Affirmatively, NC-conditioned medium (NCCM) exerts regenerative effects on CLC proliferation and extracellular matrix (ECM) production. The aim of this study was to identify NC-secreted substances that stimulate IVD regeneration. By mass spectrometry of porcine, canine and human NCCM, 149, 170 and 217 proteins were identified, respectively, with 66 proteins in common. Mainly ECM-related proteins were identified, but also organelle-derived and membrane-bound vesicle proteins. To determine whether the effect of NCCM was mediated by soluble and/or pelletable factors, porcine and canine NCCM were separated into a soluble (NCCM-S; peptides and proteins) and pelletable (NCCM-P; protein aggregates and extracellular vesicles) fraction by ultracentrifugation, and tested on bovine and canine CLCs in vitro, respectively. In each model, NCCM-S exerted a more pronounced anabolic effect than NCCM-P. However, glycosaminoglycan (GAG) uptake from the medium into the carrier gel prevented more definite conclusions. While the effect of porcine NCCM-P on bovine CLCs was negligible, canine NCCM-P appeared to enhance GAG and collagen type II deposition by canine CLCs. In conclusion, porcine and canine NCCM exerted their anabolic effects mainly through soluble factors, but also the pelletable NCCM factors showed moderate regenerative potential. Although the regenerative potential of NCCM-P should not be overlooked, future studies should focus on unraveling the protein-based regenerative mechanism from NCCM produced from isolated NCs, e.g. by NCCM fractionation and pathway blocking studies.",
author = "F.C. Bach and {de Vries}, S.A.H. and F.M. Riemers and J. Boere and {van Heel}, F.W.M. and {van Doeselaar}, M. and S.S. Goerdayal and P.G.J. Nikkels and K. Benz and Creemers, {L. B.} and A.F.M. Altelaar and B.P. Meij and K. Ito and M.A. Tryfonidou",
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Bach, FC, de Vries, SAH, Riemers, FM, Boere, J, van Heel, FWM, van Doeselaar, M, Goerdayal, SS, Nikkels, PGJ, Benz, K, Creemers, LB, Altelaar, AFM, Meij, BP, Ito, K & Tryfonidou, MA 2016, 'Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration', European Cells and Materials, vol. 32, pp. 163-180. https://doi.org/10.22203/eCM.v032a11

Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration. / Bach, F.C.; de Vries, S.A.H.; Riemers, F.M.; Boere, J.; van Heel, F.W.M. ; van Doeselaar, M.; Goerdayal, S.S.; Nikkels, P.G.J.; Benz, K.; Creemers, L. B.; Altelaar, A.F.M.; Meij, B.P.; Ito, K.; Tryfonidou, M.A.

In: European Cells and Materials, Vol. 32, 2016, p. 163-180.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Soluble and pelletable factors in porcine, canine and human notochordal cell-conditioned medium: implications for IVD regeneration

AU - Bach, F.C.

AU - de Vries, S.A.H.

AU - Riemers, F.M.

AU - Boere, J.

AU - van Heel, F.W.M.

AU - van Doeselaar, M.

AU - Goerdayal, S.S.

AU - Nikkels, P.G.J.

AU - Benz, K.

AU - Creemers, L. B.

AU - Altelaar, A.F.M.

AU - Meij, B.P.

AU - Ito, K.

AU - Tryfonidou, M.A.

PY - 2016

Y1 - 2016

N2 - During intervertebral disc (IVD) maturation, notochordal cells (NCs) are replaced by chondrocyte-like cells (CLCs) in the nucleus pulposus, suggesting that NCs play a role in maintaining tissue health. Affirmatively, NC-conditioned medium (NCCM) exerts regenerative effects on CLC proliferation and extracellular matrix (ECM) production. The aim of this study was to identify NC-secreted substances that stimulate IVD regeneration. By mass spectrometry of porcine, canine and human NCCM, 149, 170 and 217 proteins were identified, respectively, with 66 proteins in common. Mainly ECM-related proteins were identified, but also organelle-derived and membrane-bound vesicle proteins. To determine whether the effect of NCCM was mediated by soluble and/or pelletable factors, porcine and canine NCCM were separated into a soluble (NCCM-S; peptides and proteins) and pelletable (NCCM-P; protein aggregates and extracellular vesicles) fraction by ultracentrifugation, and tested on bovine and canine CLCs in vitro, respectively. In each model, NCCM-S exerted a more pronounced anabolic effect than NCCM-P. However, glycosaminoglycan (GAG) uptake from the medium into the carrier gel prevented more definite conclusions. While the effect of porcine NCCM-P on bovine CLCs was negligible, canine NCCM-P appeared to enhance GAG and collagen type II deposition by canine CLCs. In conclusion, porcine and canine NCCM exerted their anabolic effects mainly through soluble factors, but also the pelletable NCCM factors showed moderate regenerative potential. Although the regenerative potential of NCCM-P should not be overlooked, future studies should focus on unraveling the protein-based regenerative mechanism from NCCM produced from isolated NCs, e.g. by NCCM fractionation and pathway blocking studies.

AB - During intervertebral disc (IVD) maturation, notochordal cells (NCs) are replaced by chondrocyte-like cells (CLCs) in the nucleus pulposus, suggesting that NCs play a role in maintaining tissue health. Affirmatively, NC-conditioned medium (NCCM) exerts regenerative effects on CLC proliferation and extracellular matrix (ECM) production. The aim of this study was to identify NC-secreted substances that stimulate IVD regeneration. By mass spectrometry of porcine, canine and human NCCM, 149, 170 and 217 proteins were identified, respectively, with 66 proteins in common. Mainly ECM-related proteins were identified, but also organelle-derived and membrane-bound vesicle proteins. To determine whether the effect of NCCM was mediated by soluble and/or pelletable factors, porcine and canine NCCM were separated into a soluble (NCCM-S; peptides and proteins) and pelletable (NCCM-P; protein aggregates and extracellular vesicles) fraction by ultracentrifugation, and tested on bovine and canine CLCs in vitro, respectively. In each model, NCCM-S exerted a more pronounced anabolic effect than NCCM-P. However, glycosaminoglycan (GAG) uptake from the medium into the carrier gel prevented more definite conclusions. While the effect of porcine NCCM-P on bovine CLCs was negligible, canine NCCM-P appeared to enhance GAG and collagen type II deposition by canine CLCs. In conclusion, porcine and canine NCCM exerted their anabolic effects mainly through soluble factors, but also the pelletable NCCM factors showed moderate regenerative potential. Although the regenerative potential of NCCM-P should not be overlooked, future studies should focus on unraveling the protein-based regenerative mechanism from NCCM produced from isolated NCs, e.g. by NCCM fractionation and pathway blocking studies.

U2 - 10.22203/eCM.v032a11

DO - 10.22203/eCM.v032a11

M3 - Article

VL - 32

SP - 163

EP - 180

JO - European Cells and Materials

JF - European Cells and Materials

SN - 1473-2262

ER -