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Modeling Osteoarthritis Using Three-Dimensional Culture

Alexandra Fiederlein, Jodi F. Evans
American Journal of Biomedical Research. 2020, 8(3), 72-74. DOI: 10.12691/ajbr-8-3-3
Received September 01, 2020; Revised October 03, 2020; Accepted October 12, 2020

Abstract

Osteoarthritis (OA) is characterized by the degradation of cartilage caused by dysregulated and inappropriate anabolic and catabolic responses. To study this degeneration in vitro, three-dimensional (3D) culture provides a better model of the in vivo environment compared to the 2D culture along with a multitude of other benefits, providing greater insight into the progression of OA. These 3D cultures can be used in inflammation modeling of OA, demonstrating the effect of inflammatory agents such as cytokines or macrophages. In 3D damage modeling of OA, a wide variety of factors are used to cause damage, and researchers may observe repair and the progression of damage. Therapeutic modeling of OA using 3D culture allows researchers to test possible therapeutic modalities. This review highlights the benefits of 3D culture, and how, when used in inflammation, damage, and therapeutic models, brings a deeper understanding of OA.

1. Introduction

Chondrocytes and other cell types are cultured and studied in 3D due to the advantages it provides over 2D culture 1, 2, 3, 4, 5. One key factor in culturing chondrocytes in 3D is that the culture environment closely mimics the in vivo environment 1. The three-dimensional aspects of the in vivo environment, including cell-cell interactions and cell-ECM interactions, can be recapitulated in a 3D culture 6, 7. Chondrocytes in 3D culture maintain their phenotypic characteristics, such as cell shape and morphology 2, 6, 8. De-differentiation is inhibited while re-differentiation is supported 1, 2, 9, 10. The study of OA using 3D culture is not limited to chondrocytes; a variety of cell types are used, including stem cells, stromal cells, macrophages, and other chondrogenic progenitors, to investigate chondrogenesis and other processes related its progression 1, 4, 8, 11, 12, 13, 14, 15, 16. To study OA, 3D culture has been used to develop models that simulate inflammation, damage, and even therapeutic effects.

2. Discussion

2.1. Inflammation Models

Studies using 3D cultures to focus on the assessment of inflammatory agents in OA can be considered inflammatory models. Inflammatory agents involved the progression of OA include IL-1β, IL-6, IL-8, TNF-α, and NO 11, 17, 18. Inflammation models are used in a variety of ways. For example, chondrocytes derived from both osteoarthritic and normal cartilage can be co-cultured with macrophages expressing different phenotypes, activated or naive, to better understand the paracrine interactions between the two cell types 11. Transcript and protein expression levels of inflammatory markers such as IL-1β, IL-6, TNF-α, and NOS-2 can subsequently be measured. Results from these studies not only confirm other researchers’ in vitro culture results, they expand current knowledge of paracrine interactions, a benefit of using 3D culture 11. Another study found that an osmolarity of 380 mOsm in osteoarthritic chondrocytes lowers IL-6 levels, and cytokine levels in general were lower in 3D culture compared to 2D culture 18. The implementation of 3D culture in inflammation models strengthens results and provides new insights into OA 11, 17, 18.

2.2. Damage Models

Chondrocytes grown in 3D cultures that investigate the cause of damage or degradation of cartilage in OA can be considered damage models. Damage models may incorporate inflammatory agents (ex. IL-1β, TNF-α) to induce inflammation and therefore damage, 1, 12 may induce mechanical stress, enzymatic degradation, or lesions 19, 20. In co-cultures macrophage activation/ deactivation can also cause damage 11, 16. As discussed above, macrophages are used in 3D co-culture as inflammatory agents to study paracrine interactions 11. Similarly, activated macrophages can be used in 3D hydrogel cultures to study damage and hypertrophic responses. Peck and Wang found that activated macrophages induce greater hypertrophic responses than non-activated macrophages, as well as cause degradation of collagen II 16. Inflammatory cytokines such as IL-1β and TNF-α are also used to induce damage in an in vitro OA model using scaffold-free 3D cartilage transplants 12. Evans et al. (2020) are working to develop a 3D model using mesenchymal stem cells differentiated in a hyaluron matrix. Once chondrogenesis has been induced, the pellets are subjected to hyaluronidase to induce damage and the effects of stress hormones on degradation after damage are examined [personal communication]. Compared to other studies, no other studies used hyaluronidase to induce damage in a mesenchymal stem cell differentiation model to determine the effect of stress hormones on chondrogenic phenotype, giving hope for new insights on osteoarthritis. Damage models provide important information concerning the pathology of OA and preliminary models for future studies 1, 11, 12, 16, 19, 20.

2.3. Therapeutic Models

Studies using 3D cultures involving the experimentation of new or potential therapeutic advances in OA are considered therapeutic models. Therapeutic models can range from testing potassium treatments, 21 possible gene therapy, 22 testing different types of 3D culture and expansion/maintenance methods, 4, 13, 14, 15, 19, 23 to using different compounds, chemicals or drugs to assess chondrogenesis or chondrogenic repair 6, 10, 14, 15, 23. A recent study found that osteoarthritic chondrocytes, grown in 3D hydrogel and treated with hyperosmolar potassium (80 mM K+ gluconate) increased transcription of anabolic chondrogenic markers, such as aggrecan, collagen II, and SOX-9 demonstrating the therapeutic potential of this treatment 21. The potential for gene therapy has also been demonstrated using 3D culture. Transfection of a vector carrying the sequence for SOX-9 induced increased collagen II and proteoglycan transcription and reduced the hypertrophic phenotype in a 3D model of OA 22.

Using a 3D model, it is also possible to examine the efficacy of therapeutic compounds. For example, the phytoestrogen, Daidzein, has been studied in 3D culture to test its effect on the developing chondrogenic phenotype and on common chondrogenic markers (collagen II, GAG, SOX-9) 6. Chondrocytes treated with 20 µg/mL Daidzein in 3D culture had higher GAG content and collagen production than the 2D culture 6. Other studies have focused on optimizing or testing new ways to culture chondrocytes or other cell types in 3D in preparation for testing therapeutics. For example, researchers developed hyaluronic acid/human adipose-derived stem cell (hADSC) hydrogels enriched with fibrin, which promoted higher gene expression levels of collagen II and aggrecan 15. New therapeutic models using 3D culture show great promise for advances in OA treatments 4, 6, 10, 13, 14, 19, 21, 22, 23.

3. Conclusion

The use of 3D culture for modeling can provide a vast amount of information about many diseases, particularly OA, due to its versatility and applicability. It can be used to create inflammation, damage, and therapeutic models to emulate the in vivo environment more accurately when compared to 2D culture. It is important to continue developing new models of OA using 3D culture. One group, Evans et al. (2020) as highlighted above, is developing a damage model using chondrocytes derived from mesenchymal stem cell differentiation. Most work with mesenchymal stem cells has the goal of using them as a therapeutic or in repair. Using them in a new way, such as in a damage model, may provide new insights into the complex pathophysiology of OA. Overall, 3D modeling allows for advances in our understanding of the pathogenesis of OA and provides a defined in vitro system for testing the efficacy of potential therapeutics.

Acknowledgements

This work was supported by a Collegiate Science and Technology Entry Program grant to AF.

References

[1]  Galuzzi M, Perteghella S, Antonioli B, Tosca MC, Bari E, Tripodo G, Sorrenti M, Catenacci L, Mastracci L, Grillo F, et al. Human Engineered Cartilage and Decellularized Matrix as an Alternative to Animal Osteoarthritis Model. Polymers.; 10(7): 738.
In article      View Article  PubMed
 
[2]  Smeriglio P, Lai JH, Dhulipala L, Behn AW, Goodman SB, Smith RL, Maloney WJ, Yang F, Bhutani N. Comparative Potential of Juvenile and Adult Human Articular Chondrocytes for Cartilage Tissue Formation in Three-Dimensional Biomimetic Hydrogels. Tissue Engineering Part A.; 21(1-2): 147-155.
In article      View Article  PubMed
 
[3]  Erndt-Marino J, Diaz-Rodriguez P, Hahn MS. Initial In Vitro Development of a Potassium-Based Intra-Articular Injection for Osteoarthritis. Tissue Engineering Part A. 2018; 24(17-18): 1390-1392.
In article      View Article  PubMed
 
[4]  Khurshid M, Mulet‐Sierra A, Adesida A, Sen A. Osteoarthritic human chondrocytes proliferate in 3D co-culture with mesenchymal stem cells in suspension bioreactors. Journal of Tissue Engineering and Regenerative Medicine. 2018: 12(3).
In article      View Article  PubMed
 
[5]  Liu S, Xie R, Yao H, Wang D, Ren L. A biomimetic three-dimensional cartilaginous model system in vitro forosteoarthritis and osteoarthritic drug evaluation. Osteoarthritis and Cartilage. 2020; 28: S217.
In article      View Article
 
[6]  Mahmod SA, Snigh S, Djordjevic I, Mei Yee Y, Yusof R, Ramasamy TS, Rothan HA. Phytoestrogen (Daidzein) Promotes Chondrogenic Phenotype of Human Chondrocytes in 2D and 3D Culture Systems. Tissue Engineering and Regenerative Medicine. 2017 14(2): 103-112.
In article      View Article  PubMed
 
[7]  Samvelyan HJ, Hughes D, Stevens C, Staines KA. Models of Osteoarthritis: Relevance and New Insights. Calcified Tissue International. 2020 Feb 15.
In article      View Article  PubMed
 
[8]  Santos VH, Pfeifer JPH, de Souza JB, Milani BHG, de Oliveira RA, Assis MG, Deffune E, Moroz A, Alves ALG. Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules. BMC Veterinary Research. 2018; 14(1): 114.
In article      View Article  PubMed
 
[9]  Ongchai S, Somnoo O, Kongdang P, Peansukmanee S, Tangyuenyong S. TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes. Journal of Veterinary Science. 2018; 19(6): 735.
In article      View Article  PubMed
 
[10]  Varela-Eirín M, Varela-Vázquez A, Paíno CL, Casado-Díaz A, Continente AC, Mato V, Fonseca E, Kandouz M, Blanco A, Caeiro JR, et al. Senolytic activity of small molecular polyphenols from olive restores chondrocyte redifferentiation and cartilage regeneration in osteoarthritis. Cell Biology; 2019.
In article      View Article
 
[11]  Samavedi S, Diaz-Rodriguez P, Erndt-Marino JD, Hahn MS. A Three-Dimensional Chondrocyte-Macrophage Coculture System to Probe Inflammation in Experimental Osteoarthritis. Tissue Engineering Part A. 2017; 23(3-4): 101-114.
In article      View Article  PubMed
 
[12]  Weber M-C, Ponomarev I, Gaber T, Buttgereit F, Lang A. Simulating osteoarthritis in vitro with human scaffold-free 3D cartilage transplants. Osteoarthritis and Cartilage.
In article      
 
[13]  Watts AE, Ackerman-Yost JC, Nixon AJ. A Comparison of Three-Dimensional Culture Systems to Evaluate In Vitro Chondrogenesis of Equine Bone Marrow-Derived Mesenchymal Stem Cells. Tissue Engineering Part A. 2013; 19(19-20): 2275-2283.
In article      View Article  PubMed
 
[14]  Amann E, Wolff P, Breel E, van Griensven M, Balmayor ER. Hyaluronic acid facilitates chondrogenesis and matrix deposition of human adipose derived mesenchymal stem cells and human chondrocytes co-cultures. Acta Biomaterialia. 2017; 52: 130-144.
In article      View Article  PubMed
 
[15]  Wu S-C, Huang P-Y, Chen C-H, Teong B, Chen J-W, Wu C-W, Chang J-K, Ho M-L. Hyaluronan microenvironment enhances cartilage regeneration of human adipose-derived stem cells in a chondral defect model. International Journal of Biological Macromolecules. 2018]; 119: 726-740.
In article      View Article  PubMed
 
[16]  Peck Y, Wang D. An engineered biomimetic cartilaginous tissue model for osteoarthritis drug evaluation. Osteoarthritis and Cartilage. 2015 23: A413-A414.
In article      View Article
 
[17]  Hikida M, Kanamoto T, Hanai T, Yo J, Sato S, Nakata K. Cyclic compression loading on three-dimensional tissue of human articular chondrocytes upregulates inflammatory mediators and pain-sensitizing molecules. Osteoarthritis and Cartilage. 2019; 27: S417-S418.
In article      View Article
 
[18]  Mang T, Lindemann S, Gigout A. Increasing the Medium Osmolarity Reduces the Inflammatory Status of Human OA Chondrocytes and Increases Their Responsiveness to GDF-5. International Journal of Molecular Sciences. 2020; 21(2): 531.
In article      View Article  PubMed
 
[19]  Sanjurjo-Rodríguez C, Castro-Viñuelas R, Hermida-Gómez T, Fuentes-Boquete IM, de Toro FJ, Blanco FJ, Díaz-Prado SM. Human Cartilage Engineering in an In Vitro Repair Model Using Collagen Scaffolds and Mesenchymal Stromal Cells. International Journal of Medical Sciences. 2017; 14(12): 1257-1262.
In article      View Article  PubMed
 
[20]  Jung Y-K, Shin D, Park D, Kim J, Han S. Breakdown of the extracellular matrix recapitulates osteoarthritic phenotypes in 3d-hydrogel model: the validation of in vitro cell culture model of osteoarthritis. Osteoarthritis and Cartilage. 2020; 28: S186.
In article      View Article
 
[21]  Erndt-Marino J, Trinkle E, Hahn MS. Hyperosmolar Potassium (K +) Treatment Suppresses Osteoarthritic Chondrocyte Catabolic and Inflammatory Protein Production in a 3-Dimensional In Vitro Model. CARTILAGE. 2019; 10(2): 186-195.
In article      View Article  PubMed
 
[22]  Daniels O, Frisch J, Venkatesan JK, Rey-Rico A, Schmitt G, Cucchiarini M. Effects of rAAV-Mediated sox9 Overexpression on the Biological Activities of Human Osteoarthritic Articular Chondrocytes in Their Intrinsic Three-Dimensional Environment. Journal of Clinical Medicine. 2019; 8(10): 1637.
In article      View Article  PubMed
 
[23]  La Gatta A, Ricci G, Stellavato A, Cammarota M, Filosa R, Papa A, D’Agostino A, Portaccio M, Delfino I, De Rosa M, et al. Hyaluronan hydrogels with a low degree of modification as scaffolds for cartilage engineering. International Journal of Biological Macromolecules. 2017; 103: 978-989.
In article      View Article  PubMed
 

Published with license by Science and Education Publishing, Copyright © 2020 Alexandra Fiederlein and Jodi F. Evans

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Cite this article:

Normal Style
Alexandra Fiederlein, Jodi F. Evans. Modeling Osteoarthritis Using Three-Dimensional Culture. American Journal of Biomedical Research. Vol. 8, No. 3, 2020, pp 72-74. http://pubs.sciepub.com/ajbr/8/3/3
MLA Style
Fiederlein, Alexandra, and Jodi F. Evans. "Modeling Osteoarthritis Using Three-Dimensional Culture." American Journal of Biomedical Research 8.3 (2020): 72-74.
APA Style
Fiederlein, A. , & Evans, J. F. (2020). Modeling Osteoarthritis Using Three-Dimensional Culture. American Journal of Biomedical Research, 8(3), 72-74.
Chicago Style
Fiederlein, Alexandra, and Jodi F. Evans. "Modeling Osteoarthritis Using Three-Dimensional Culture." American Journal of Biomedical Research 8, no. 3 (2020): 72-74.
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[1]  Galuzzi M, Perteghella S, Antonioli B, Tosca MC, Bari E, Tripodo G, Sorrenti M, Catenacci L, Mastracci L, Grillo F, et al. Human Engineered Cartilage and Decellularized Matrix as an Alternative to Animal Osteoarthritis Model. Polymers.; 10(7): 738.
In article      View Article  PubMed
 
[2]  Smeriglio P, Lai JH, Dhulipala L, Behn AW, Goodman SB, Smith RL, Maloney WJ, Yang F, Bhutani N. Comparative Potential of Juvenile and Adult Human Articular Chondrocytes for Cartilage Tissue Formation in Three-Dimensional Biomimetic Hydrogels. Tissue Engineering Part A.; 21(1-2): 147-155.
In article      View Article  PubMed
 
[3]  Erndt-Marino J, Diaz-Rodriguez P, Hahn MS. Initial In Vitro Development of a Potassium-Based Intra-Articular Injection for Osteoarthritis. Tissue Engineering Part A. 2018; 24(17-18): 1390-1392.
In article      View Article  PubMed
 
[4]  Khurshid M, Mulet‐Sierra A, Adesida A, Sen A. Osteoarthritic human chondrocytes proliferate in 3D co-culture with mesenchymal stem cells in suspension bioreactors. Journal of Tissue Engineering and Regenerative Medicine. 2018: 12(3).
In article      View Article  PubMed
 
[5]  Liu S, Xie R, Yao H, Wang D, Ren L. A biomimetic three-dimensional cartilaginous model system in vitro forosteoarthritis and osteoarthritic drug evaluation. Osteoarthritis and Cartilage. 2020; 28: S217.
In article      View Article
 
[6]  Mahmod SA, Snigh S, Djordjevic I, Mei Yee Y, Yusof R, Ramasamy TS, Rothan HA. Phytoestrogen (Daidzein) Promotes Chondrogenic Phenotype of Human Chondrocytes in 2D and 3D Culture Systems. Tissue Engineering and Regenerative Medicine. 2017 14(2): 103-112.
In article      View Article  PubMed
 
[7]  Samvelyan HJ, Hughes D, Stevens C, Staines KA. Models of Osteoarthritis: Relevance and New Insights. Calcified Tissue International. 2020 Feb 15.
In article      View Article  PubMed
 
[8]  Santos VH, Pfeifer JPH, de Souza JB, Milani BHG, de Oliveira RA, Assis MG, Deffune E, Moroz A, Alves ALG. Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules. BMC Veterinary Research. 2018; 14(1): 114.
In article      View Article  PubMed
 
[9]  Ongchai S, Somnoo O, Kongdang P, Peansukmanee S, Tangyuenyong S. TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes. Journal of Veterinary Science. 2018; 19(6): 735.
In article      View Article  PubMed
 
[10]  Varela-Eirín M, Varela-Vázquez A, Paíno CL, Casado-Díaz A, Continente AC, Mato V, Fonseca E, Kandouz M, Blanco A, Caeiro JR, et al. Senolytic activity of small molecular polyphenols from olive restores chondrocyte redifferentiation and cartilage regeneration in osteoarthritis. Cell Biology; 2019.
In article      View Article
 
[11]  Samavedi S, Diaz-Rodriguez P, Erndt-Marino JD, Hahn MS. A Three-Dimensional Chondrocyte-Macrophage Coculture System to Probe Inflammation in Experimental Osteoarthritis. Tissue Engineering Part A. 2017; 23(3-4): 101-114.
In article      View Article  PubMed
 
[12]  Weber M-C, Ponomarev I, Gaber T, Buttgereit F, Lang A. Simulating osteoarthritis in vitro with human scaffold-free 3D cartilage transplants. Osteoarthritis and Cartilage.
In article      
 
[13]  Watts AE, Ackerman-Yost JC, Nixon AJ. A Comparison of Three-Dimensional Culture Systems to Evaluate In Vitro Chondrogenesis of Equine Bone Marrow-Derived Mesenchymal Stem Cells. Tissue Engineering Part A. 2013; 19(19-20): 2275-2283.
In article      View Article  PubMed
 
[14]  Amann E, Wolff P, Breel E, van Griensven M, Balmayor ER. Hyaluronic acid facilitates chondrogenesis and matrix deposition of human adipose derived mesenchymal stem cells and human chondrocytes co-cultures. Acta Biomaterialia. 2017; 52: 130-144.
In article      View Article  PubMed
 
[15]  Wu S-C, Huang P-Y, Chen C-H, Teong B, Chen J-W, Wu C-W, Chang J-K, Ho M-L. Hyaluronan microenvironment enhances cartilage regeneration of human adipose-derived stem cells in a chondral defect model. International Journal of Biological Macromolecules. 2018]; 119: 726-740.
In article      View Article  PubMed
 
[16]  Peck Y, Wang D. An engineered biomimetic cartilaginous tissue model for osteoarthritis drug evaluation. Osteoarthritis and Cartilage. 2015 23: A413-A414.
In article      View Article
 
[17]  Hikida M, Kanamoto T, Hanai T, Yo J, Sato S, Nakata K. Cyclic compression loading on three-dimensional tissue of human articular chondrocytes upregulates inflammatory mediators and pain-sensitizing molecules. Osteoarthritis and Cartilage. 2019; 27: S417-S418.
In article      View Article
 
[18]  Mang T, Lindemann S, Gigout A. Increasing the Medium Osmolarity Reduces the Inflammatory Status of Human OA Chondrocytes and Increases Their Responsiveness to GDF-5. International Journal of Molecular Sciences. 2020; 21(2): 531.
In article      View Article  PubMed
 
[19]  Sanjurjo-Rodríguez C, Castro-Viñuelas R, Hermida-Gómez T, Fuentes-Boquete IM, de Toro FJ, Blanco FJ, Díaz-Prado SM. Human Cartilage Engineering in an In Vitro Repair Model Using Collagen Scaffolds and Mesenchymal Stromal Cells. International Journal of Medical Sciences. 2017; 14(12): 1257-1262.
In article      View Article  PubMed
 
[20]  Jung Y-K, Shin D, Park D, Kim J, Han S. Breakdown of the extracellular matrix recapitulates osteoarthritic phenotypes in 3d-hydrogel model: the validation of in vitro cell culture model of osteoarthritis. Osteoarthritis and Cartilage. 2020; 28: S186.
In article      View Article
 
[21]  Erndt-Marino J, Trinkle E, Hahn MS. Hyperosmolar Potassium (K +) Treatment Suppresses Osteoarthritic Chondrocyte Catabolic and Inflammatory Protein Production in a 3-Dimensional In Vitro Model. CARTILAGE. 2019; 10(2): 186-195.
In article      View Article  PubMed
 
[22]  Daniels O, Frisch J, Venkatesan JK, Rey-Rico A, Schmitt G, Cucchiarini M. Effects of rAAV-Mediated sox9 Overexpression on the Biological Activities of Human Osteoarthritic Articular Chondrocytes in Their Intrinsic Three-Dimensional Environment. Journal of Clinical Medicine. 2019; 8(10): 1637.
In article      View Article  PubMed
 
[23]  La Gatta A, Ricci G, Stellavato A, Cammarota M, Filosa R, Papa A, D’Agostino A, Portaccio M, Delfino I, De Rosa M, et al. Hyaluronan hydrogels with a low degree of modification as scaffolds for cartilage engineering. International Journal of Biological Macromolecules. 2017; 103: 978-989.
In article      View Article  PubMed