Differential Production of Cartilage ECM in 3D Agarose Constructs by Equine Articular Cartilage Progenitor Cells and Mesenchymal Stromal Cells.
Abstract: Identification of articular cartilage progenitor cells (ACPCs) has opened up new opportunities for cartilage repair. These cells may be used as alternatives for or in combination with mesenchymal stromal cells (MSCs) in cartilage engineering. However, their potential needs to be further investigated, since only a few studies have compared ACPCs and MSCs when cultured in hydrogels. Therefore, in this study, we compared chondrogenic differentiation of equine ACPCs and MSCs in agarose constructs as monocultures and as zonally layered co-cultures under both normoxic and hypoxic conditions. ACPCs and MSCs exhibited distinctly differential production of the cartilaginous extracellular matrix (ECM). For ACPC constructs, markedly higher glycosaminoglycan (GAG) contents were determined by histological and quantitative biochemical evaluation, both in normoxia and hypoxia. Differential GAG production was also reflected in layered co-culture constructs. For both cell types, similar staining for type II collagen was detected. However, distinctly weaker staining for undesired type I collagen was observed in the ACPC constructs. For ACPCs, only very low alkaline phosphatase (ALP) activity, a marker of terminal differentiation, was determined, in stark contrast to what was found for MSCs. This study underscores the potential of ACPCs as a promising cell source for cartilage engineering.
Publication Date: 2020-09-25 PubMed ID: 32992847PubMed Central: PMC7582568DOI: 10.3390/ijms21197071Google Scholar: Lookup
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Summary
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This research study explores the potential of Articular Cartilage Progenitor Cells (ACPCs) as a viable source for cartilage repair. The study specifically compares the chondrogenic differentiation and production of cartilaginous extracellular matrix (ECM) of ACPCs and Mesenchymal Stromal Cells (MSCs) when cultured in gel-like structures.
Origins and Aims
- The discovery of ACPCs has presented new possibilities in term of cartilage repair. The study therefore aims to explore these potentials, particularly conccerning their use alone or alongside MSCs for cartilage engineering.
- In this research, equine ACPCs and MSCs were cultured in three-dimensional agarose constructs, both separately and together in co-cultures, under both normal and low oxygen conditions.
Findings and Results
- Both ACPCs and MSCs produced distinctly different amounts of ECM. Of the two, ACPC constructs had noticeably higher amounts of glycosaminoglycan (GAG), critical for the support and binding of cells in the cartilage.
- The high levels of GAG were consistent in both normal and low oxygen conditions and the co-cultures reflected similar GAG production patterns.
- Both cell types had similar staining results for type II collagen, a protein found in human cartilage. However, ACPCs showed remarkably less staining for type I collagen, which is not typically found in cartilage and is therefore undesirable.
Implications
- ACPC constructs displayed only low levels of Alkaline Phosphatase (ALP) activity. ALP is a marker of terminal differentiation, the process by which cells become specialised, and high levels could indicate potential cellular senescence, or aging. The contrastingly high readings for MSCs signified a pronounced divergence between the two cell types.
- The research emphasizes the potential of ACPCs as a promising option for cartilage engineering. This finding could have lasting implications in the field of human medicine, particularly concerning treatments for damaged or degenerating cartilage, such as in cases of arthritis.
Cite This Article
APA
Schmidt S, Abinzano F, Mensinga A, Teßmar J, Groll J, Malda J, Levato R, Blunk T.
(2020).
Differential Production of Cartilage ECM in 3D Agarose Constructs by Equine Articular Cartilage Progenitor Cells and Mesenchymal Stromal Cells.
Int J Mol Sci, 21(19), 7071.
https://doi.org/10.3390/ijms21197071 Publication
Researcher Affiliations
- Department of Trauma, Hand, Plastic and Reconstructive Surgery, University of Würzburg, Oberdürrbacher Str. 6, 97080 Würzburg, Germany.
- Department of Orthopedics, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands.
- Department of Orthopedics, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands.
- Department for Functional Materials in Medicine and Dentistry and Bavarian Polymer Institute, University of Würzburg, Pleicherwall 2, 97070 Würzburg, Germany.
- Department for Functional Materials in Medicine and Dentistry and Bavarian Polymer Institute, University of Würzburg, Pleicherwall 2, 97070 Würzburg, Germany.
- Department of Orthopedics, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands.
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 CM Utrecht, The Netherlands.
- Department of Orthopedics, University Medical Center Utrecht, 3508 GA Utrecht, The Netherlands.
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 CM Utrecht, The Netherlands.
- Department of Trauma, Hand, Plastic and Reconstructive Surgery, University of Würzburg, Oberdürrbacher Str. 6, 97080 Würzburg, Germany.
MeSH Terms
- Animals
- Cartilage, Articular / cytology
- Cell Differentiation
- Cells, Cultured
- Chondrogenesis
- Horses
- Mesenchymal Stem Cells / cytology
- Stem Cells / cytology
- Tissue Engineering
Grant Funding
- 309962 / Seventh Framework Programme
- 326998133 (SFB TRR 225) / Deutsche Forschungsgemeinschaft
Conflict of Interest Statement
The authors declare no conflict of interest.
References
This article includes 68 references
- Caldwell KL, Wang J. Cell-based articular cartilage repair: the link between development and regeneration.. Osteoarthritis Cartilage 2015 Mar;23(3):351-62.
- Richter DL, Schenck RC Jr, Wascher DC, Treme G. Knee Articular Cartilage Repair and Restoration Techniques: A Review of the Literature.. Sports Health 2016 Mar-Apr;8(2):153-60.
- Schnabel M, Marlovits S, Eckhoff G, Fichtel I, Gotzen L, Vécsei V, Schlegel J. Dedifferentiation-associated changes in morphology and gene expression in primary human articular chondrocytes in cell culture.. Osteoarthritis Cartilage 2002 Jan;10(1):62-70.
- Caron MM, Emans PJ, Coolsen MM, Voss L, Surtel DA, Cremers A, van Rhijn LW, Welting TJ. Redifferentiation of dedifferentiated human articular chondrocytes: comparison of 2D and 3D cultures.. Osteoarthritis Cartilage 2012 Oct;20(10):1170-8.
- Im GI. Tissue Engineering in Osteoarthritis: Current Status and Prospect of Mesenchymal Stem Cell Therapy.. BioDrugs 2018 Jun;32(3):183-192.
- Somoza RA, Welter JF, Correa D, Caplan AI. Chondrogenic differentiation of mesenchymal stem cells: challenges and unfulfilled expectations.. Tissue Eng Part B Rev 2014 Dec;20(6):596-608.
- Pelttari K, Winter A, Steck E, Goetzke K, Hennig T, Ochs BG, Aigner T, Richter W. Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice.. Arthritis Rheum 2006 Oct;54(10):3254-66.
- Mueller MB, Tuan RS. Functional characterization of hypertrophy in chondrogenesis of human mesenchymal stem cells.. Arthritis Rheum 2008 May;58(5):1377-88.
- Dowthwaite GP, Bishop JC, Redman SN, Khan IM, Rooney P, Evans DJ, Haughton L, Bayram Z, Boyer S, Thomson B, Wolfe MS, Archer CW. The surface of articular cartilage contains a progenitor cell population.. J Cell Sci 2004 Feb 29;117(Pt 6):889-97.
- Alsalameh S, Amin R, Gemba T, Lotz M. Identification of mesenchymal progenitor cells in normal and osteoarthritic human articular cartilage.. Arthritis Rheum 2004 May;50(5):1522-32.
- Khan IM, Bishop JC, Gilbert S, Archer CW. Clonal chondroprogenitors maintain telomerase activity and Sox9 expression during extended monolayer culture and retain chondrogenic potential.. Osteoarthritis Cartilage 2009 Apr;17(4):518-28.
- Williams R, Khan IM, Richardson K, Nelson L, McCarthy HE, Analbelsi T, Singhrao SK, Dowthwaite GP, Jones RE, Baird DM, Lewis H, Roberts S, Shaw HM, Dudhia J, Fairclough J, Briggs T, Archer CW. Identification and clonal characterisation of a progenitor cell sub-population in normal human articular cartilage.. PLoS One 2010 Oct 14;5(10):e13246.
- McCarthy HE, Bara JJ, Brakspear K, Singhrao SK, Archer CW. The comparison of equine articular cartilage progenitor cells and bone marrow-derived stromal cells as potential cell sources for cartilage repair in the horse.. Vet J 2012 Jun;192(3):345-51.
- Nelson L, McCarthy HE, Fairclough J, Williams R, Archer CW. Evidence of a Viable Pool of Stem Cells within Human Osteoarthritic Cartilage.. Cartilage 2014 Oct;5(4):203-14.
- Jayasuriya CT, Chen Q. Potential benefits and limitations of utilizing chondroprogenitors in cell-based cartilage therapy.. Connect Tissue Res 2015;56(4):265-71.
- Jiang Y, Cai Y, Zhang W, Yin Z, Hu C, Tong T, Lu P, Zhang S, Neculai D, Tuan RS, Ouyang HW. Human Cartilage-Derived Progenitor Cells From Committed Chondrocytes for Efficient Cartilage Repair and Regeneration.. Stem Cells Transl Med 2016 Jun;5(6):733-44.
- Frisbie DD, McCarthy HE, Archer CW, Barrett MF, McIlwraith CW. Evaluation of articular cartilage progenitor cells for the repair of articular defects in an equine model.. J Bone Joint Surg Am 2015 Mar 18;97(6):484-93.
- Mancini IAD, Schmidt S, Brommer H, Pouran B, Schäfer S, Tessmar J, Mensinga A, van Rijen MHP, Groll J, Blunk T, Levato R, Malda J, van Weeren PR. A composite hydrogel-3D printed thermoplast osteochondral anchor as example for a zonal approach to cartilage repair: in vivo performance in a long-term equine model.. Biofabrication 2020 Jul 1;12(3):035028.
- Xue K, Zhang X, Gao Z, Xia W, Qi L, Liu K. Cartilage progenitor cells combined with PHBV in cartilage tissue engineering.. J Transl Med 2019 Mar 29;17(1):104.
- Levato R, Webb WR, Otto IA, Mensinga A, Zhang Y, van Rijen M, van Weeren R, Khan IM, Malda J. The bio in the ink: cartilage regeneration with bioprintable hydrogels and articular cartilage-derived progenitor cells.. Acta Biomater 2017 Oct 1;61:41-53.
- Mouser VHM, Levato R, Mensinga A, Dhert WJA, Gawlitta D, Malda J. Bio-ink development for three-dimensional bioprinting of hetero-cellular cartilage constructs.. Connect Tissue Res 2020 Mar;61(2):137-151.
- Benya PD, Shaffer JD. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels.. Cell 1982 Aug;30(1):215-24.
- Lee DA, Bader DL. The development and characterization of an in vitro system to study strain-induced cell deformation in isolated chondrocytes.. In Vitro Cell Dev Biol Anim 1995 Dec;31(11):828-35.
- Carreau A, El Hafny-Rahbi B, Matejuk A, Grillon C, Kieda C. Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia.. J Cell Mol Med 2011 Jun;15(6):1239-53.
- Anderson DE, Markway BD, Weekes KJ, McCarthy HE, Johnstone B. Physioxia Promotes the Articular Chondrocyte-Like Phenotype in Human Chondroprogenitor-Derived Self-Organized Tissue.. Tissue Eng Part A 2018 Feb;24(3-4):264-274.
- Kim TK, Sharma B, Williams CG, Ruffner MA, Malik A, McFarland EG, Elisseeff JH. Experimental model for cartilage tissue engineering to regenerate the zonal organization of articular cartilage.. Osteoarthritis Cartilage 2003 Sep;11(9):653-64.
- Kunisch E, Knauf AK, Hesse E, Freudenberg U, Werner C, Bothe F, Diederichs S, Richter W. StarPEG/heparin-hydrogel based in vivo engineering of stable bizonal cartilage with a calcified bottom layer.. Biofabrication 2018 Oct 30;11(1):015001.
- Yin L, Wu Y, Yang Z, Denslin V, Ren X, Tee CA, Lai Z, Lim CT, Han J, Lee EH. Characterization and application of size-sorted zonal chondrocytes for articular cartilage regeneration.. Biomaterials 2018 May;165:66-78.
- Owida HA, Yang R, Cen L, Kuiper NJ, Yang Y. Induction of zonal-specific cellular morphology and matrix synthesis for biomimetic cartilage regeneration using hybrid scaffolds.. J R Soc Interface 2018 Jun;15(143).
- Zhu D, Tong X, Trinh P, Yang F. Mimicking Cartilage Tissue Zonal Organization by Engineering Tissue-Scale Gradient Hydrogels as 3D Cell Niche.. Tissue Eng Part A 2018 Jan;24(1-2):1-10.
- Ng KW, Wang CC, Mauck RL, Kelly TA, Chahine NO, Costa KD, Ateshian GA, Hung CT. A layered agarose approach to fabricate depth-dependent inhomogeneity in chondrocyte-seeded constructs.. J Orthop Res 2005 Jan;23(1):134-41.
- Castilho M, Mouser V, Chen M, Malda J, Ito K. Bi-layered micro-fibre reinforced hydrogels for articular cartilage regeneration.. Acta Biomater 2019 Sep 1;95:297-306.
- Acharya C, Adesida A, Zajac P, Mumme M, Riesle J, Martin I, Barbero A. Enhanced chondrocyte proliferation and mesenchymal stromal cells chondrogenesis in coculture pellets mediate improved cartilage formation.. J Cell Physiol 2012 Jan;227(1):88-97.
- Bian L, Zhai DY, Mauck RL, Burdick JA. Coculture of human mesenchymal stem cells and articular chondrocytes reduces hypertrophy and enhances functional properties of engineered cartilage.. Tissue Eng Part A 2011 Apr;17(7-8):1137-45.
- Cooke ME, Allon AA, Cheng T, Kuo AC, Kim HT, Vail TP, Marcucio RS, Schneider RA, Lotz JC, Alliston T. Structured three-dimensional co-culture of mesenchymal stem cells with chondrocytes promotes chondrogenic differentiation without hypertrophy.. Osteoarthritis Cartilage 2011 Oct;19(10):1210-8.
- Vinod E, Kachroo U, Sathishkumar S, Boopalan P R J V C. In vitro characterization of human articular chondrocytes and chondroprogenitors derived from normal and osteoarthritic knee joints. BioRxiv 2018:440107.
- Roberts S, Menage J, Sandell LJ, Evans EH, Richardson JB. Immunohistochemical study of collagen types I and II and procollagen IIA in human cartilage repair tissue following autologous chondrocyte implantation.. Knee 2009 Oct;16(5):398-404.
- Schizas N, Savvidou O, Triantafyllopoulos I, Papadakis S, Dontas I, Papagelopoulos P. Adjuvant therapies for the enhancement of microfracture technique in cartilage repair.. Orthop Rev (Pavia) 2019 Sep 24;11(3):7950.
- Mithoefer K, Williams RJ 3rd, Warren RF, Potter HG, Spock CR, Jones EC, Wickiewicz TL, Marx RG. Chondral resurfacing of articular cartilage defects in the knee with the microfracture technique. Surgical technique.. J Bone Joint Surg Am 2006 Sep;88 Suppl 1 Pt 2:294-304.
- Steadman JR, Briggs KK, Rodrigo JJ, Kocher MS, Gill TJ, Rodkey WG. Outcomes of microfracture for traumatic chondral defects of the knee: average 11-year follow-up.. Arthroscopy 2003 May-Jun;19(5):477-84.
- Meretoja VV, Dahlin RL, Wright S, Kasper FK, Mikos AG. The effect of hypoxia on the chondrogenic differentiation of co-cultured articular chondrocytes and mesenchymal stem cells in scaffolds.. Biomaterials 2013 Jun;34(17):4266-73.
- Mauck RL, Yuan X, Tuan RS. Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture.. Osteoarthritis Cartilage 2006 Feb;14(2):179-89.
- Huang X, Hou Y, Zhong L, Huang D, Qian H, Karperien M, Chen W. Promoted Chondrogenesis of Cocultured Chondrocytes and Mesenchymal Stem Cells under Hypoxia Using In-situ Forming Degradable Hydrogel Scaffolds.. Biomacromolecules 2018 Jan 8;19(1):94-102.
- Tiruvannamalai Annamalai R, Mertz DR, Daley EL, Stegemann JP. Collagen Type II enhances chondrogenic differentiation in agarose-based modular microtissues.. Cytotherapy 2016 Feb;18(2):263-77.
- Daly AC, Critchley SE, Rencsok EM, Kelly DJ. A comparison of different bioinks for 3D bioprinting of fibrocartilage and hyaline cartilage.. Biofabrication 2016 Oct 7;8(4):045002.
- Krouwels A, Melchels FPW, van Rijen MHP, Ten Brink CBM, Dhert WJA, Cumhur Öner F, Tryfonidou MA, Creemers LB. Focal adhesion signaling affects regeneration by human nucleus pulposus cells in collagen- but not carbohydrate-based hydrogels.. Acta Biomater 2018 Jan 15;66:238-247.
- Sophia Fox AJ, Bedi A, Rodeo SA. The basic science of articular cartilage: structure, composition, and function.. Sports Health 2009 Nov;1(6):461-8.
- Klein TJ, Malda J, Sah RL, Hutmacher DW. Tissue engineering of articular cartilage with biomimetic zones.. Tissue Eng Part B Rev 2009 Jun;15(2):143-57.
- Klein TJ, Schumacher BL, Schmidt TA, Li KW, Voegtline MS, Masuda K, Thonar EJ, Sah RL. Tissue engineering of stratified articular cartilage from chondrocyte subpopulations.. Osteoarthritis Cartilage 2003 Aug;11(8):595-602.
- Kim M, Steinberg DR, Burdick JA, Mauck RL. Extracellular vesicles mediate improved functional outcomes in engineered cartilage produced from MSC/chondrocyte cocultures.. Proc Natl Acad Sci U S A 2019 Jan 29;116(5):1569-1578.
- Zuo Q, Cui W, Liu F, Wang Q, Chen Z, Fan W. Co-cultivated mesenchymal stem cells support chondrocytic differentiation of articular chondrocytes.. Int Orthop 2013 Apr;37(4):747-52.
- Zhu M, Feng Q, Bian L. Differential effect of hypoxia on human mesenchymal stem cell chondrogenesis and hypertrophy in hyaluronic acid hydrogels.. Acta Biomater 2014 Mar;10(3):1333-40.
- Bae HC, Park HJ, Wang SY, Yang HR, Lee MC, Han HS. Hypoxic condition enhances chondrogenesis in synovium-derived mesenchymal stem cells.. Biomater Res 2018;22:28.
- Robins JC, Akeno N, Mukherjee A, Dalal RR, Aronow BJ, Koopman P, Clemens TL. Hypoxia induces chondrocyte-specific gene expression in mesenchymal cells in association with transcriptional activation of Sox9.. Bone 2005 Sep;37(3):313-22.
- Leijten J, Georgi N, Moreira Teixeira L, van Blitterswijk CA, Post JN, Karperien M. Metabolic programming of mesenchymal stromal cells by oxygen tension directs chondrogenic cell fate.. Proc Natl Acad Sci U S A 2014 Sep 23;111(38):13954-9.
- Gawlitta D, van Rijen MH, Schrijver EJ, Alblas J, Dhert WJ. Hypoxia impedes hypertrophic chondrogenesis of human multipotent stromal cells.. Tissue Eng Part A 2012 Oct;18(19-20):1957-66.
- Merceron C, Vinatier C, Portron S, Masson M, Amiaud J, Guigand L, Chérel Y, Weiss P, Guicheux J. Differential effects of hypoxia on osteochondrogenic potential of human adipose-derived stem cells.. Am J Physiol Cell Physiol 2010 Feb;298(2):C355-64.
- Desancé M, Contentin R, Bertoni L, Gomez-Leduc T, Branly T, Jacquet S, Betsch JM, Batho A, Legendre F, Audigié F, Galéra P, Demoor M. Chondrogenic Differentiation of Defined Equine Mesenchymal Stem Cells Derived from Umbilical Cord Blood for Use in Cartilage Repair Therapy.. Int J Mol Sci 2018 Feb 10;19(2).
- Anderson DE, Markway BD, Bond D, McCarthy HE, Johnstone B. Responses to altered oxygen tension are distinct between human stem cells of high and low chondrogenic capacity.. Stem Cell Res Ther 2016 Oct 20;7(1):154.
- Adesida AB, Mulet-Sierra A, Jomha NM. Hypoxia mediated isolation and expansion enhances the chondrogenic capacity of bone marrow mesenchymal stromal cells.. Stem Cell Res Ther 2012 Mar 2;3(2):9.
- Böck T, Schill V, Krähnke M, Steinert AF, Tessmar J, Blunk T, Groll J. TGF-β1-Modified Hyaluronic Acid/Poly(glycidol) Hydrogels for Chondrogenic Differentiation of Human Mesenchymal Stromal Cells.. Macromol Biosci 2018 Jul;18(7):e1700390.
- Kim YJ, Sah RL, Doong JY, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst 33258.. Anal Biochem 1988 Oct;174(1):168-76.
- WOESSNER JF Jr. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid.. Arch Biochem Biophys 1961 May;93:440-7.
- Hollander AP, Heathfield TF, Webber C, Iwata Y, Bourne R, Rorabeck C, Poole AR. Increased damage to type II collagen in osteoarthritic articular cartilage detected by a new immunoassay.. J Clin Invest 1994 Apr;93(4):1722-32.
- Farndale RW, Buttle DJ, Barrett AJ. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue.. Biochim Biophys Acta 1986 Sep 4;883(2):173-7.
- BESSEY OA, LOWRY OH, BROCK MJ. A method for the rapid determination of alkaline phosphates with five cubic millimeters of serum.. J Biol Chem 1946 Jul;164:321-9.
- Martin I, Obradovic B, Freed LE, Vunjak-Novakovic G. Method for quantitative analysis of glycosaminoglycan distribution in cultured natural and engineered cartilage.. Ann Biomed Eng 1999 Sep-Oct;27(5):656-62.
- SWEAT F, PUCHTLER H, ROSENTHAL SI. SIRIUS RED F3BA AS A STAIN FOR CONNECTIVE TISSUE.. Arch Pathol 1964 Jul;78:69-72.
Citations
This article has been cited 3 times.- Roncada T, Bonithon R, Blunn G, Roldo M. Soft substrates direct stem cell differentiation into the chondrogenic lineage without the use of growth factors.. J Tissue Eng 2022 Jan-Dec;13:20417314221122121.
- Wei Z, Zhang G, Cao Q, Zhao T, Bian Y, Zhu W, Weng X. Recent Developments and Current Applications of Organic Nanomaterials in Cartilage Repair.. Bioengineering (Basel) 2022 Aug 15;9(8).
- Rikkers M, Korpershoek JV, Levato R, Malda J, Vonk LA. Progenitor Cells in Healthy and Osteoarthritic Human Cartilage Have Extensive Culture Expansion Capacity while Retaining Chondrogenic Properties.. Cartilage 2021 Dec;13(2_suppl):129S-142S.
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