Skip to main content

Advertisement

Log in

Effects of culture conditions on the mechanical and biological properties of engineered cartilage constructed with collagen hybrid scaffold and human mesenchymal stem cells

  • Tissue Engineering Constructs and Cell Substrates
  • Original Research
  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

Mesenchymal stem cells (MSCs) has been used as one of the new cell sources in osteochondral tissue engineering. It has been well known that control of their differentiation into chondrocytes plays a key role in developing engineered cartilages. Therefore, this study aims to develop a fundamental protocol to control the differentiation and proliferation of MSCs to construct an engineered cartilage. We compared the effects of three different culture conditions on cell proliferation, extracellular matrix formation and the mechanical response of engineered cartilage constructed using a collagen-based hybrid scaffold and human MSCs. The experimental results clearly showed that the combined culture condition of the chondrogenic differentiation culture and the chondrocyte growth culture exhibited statistically significant cell proliferation, ECM formation and stiffness responses as compared to the other two combinations. It is thus concluded that the combination of the differentiation culture with the subsequent growth culture is recommended as the culture condition for chondrogenic tissue engineering using hMSCs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Lungu E, Vendittoli PA, Desmeules F. Identification of patients with suboptimal results after hip arthroplasty: development of a preliminary prediction algorithm. Bmc Musculoskelet Disord. 2015;16:279.

    Google Scholar 

  2. Kalamegam G, Memic A, Budd E, Abbas M, Mobasheri A. A comprehensive review of stem cells for cartilage regeneration in osteoarthritis. Adv Exp Med Biol. 2018;1089:23–36.

    CAS  Google Scholar 

  3. Shen G, Zhang J-F, Fang F-Z. In vitro evaluation of artificial joints: a comprehensive review. Adv Manuf. 2019;7:1–14.

    CAS  Google Scholar 

  4. Ichinose S, Tagami M, Muneta T, Mukohyama H, Sekiya I. Comparative sequential morphological analyses during in vitro chondrogenesis and osteogenesis of mesenchymal stem cells embedded in collagen gels. Med Mol Morphol. 2013;46:24–33.

    CAS  Google Scholar 

  5. Adachi N, Ochi M, Deie M, Nakamae A, Kamei G, Uchio Y. et al. Implantation of tissue-engineered cartilage-like tissue for the treatment for full-thickness cartilage defects of the knee. Knee Surg Sports Traumatol Arthrosc.2014;22:1241–8.

    Google Scholar 

  6. Mori H, Kondo E, Kawaguchi Y, Kitamura N, Nagai N, Iida H. et al. Development of a salmon-derived crosslinked atelocollagen sponge disc containing osteogenic protein-1 for articular cartilage regeneration: in vivo evaluations with rabbits. Bmc Musculoskelet Disord. 2013;14:1471–2474.

    Google Scholar 

  7. Liao J, Qu Y, Chu B, Zhang X, Qian Z. Biodegradable CSMA/PECA/Graphene porous hybrid scaffold for cartilage tissue engineering. Sci Rep. 2015;5:9879.

    CAS  Google Scholar 

  8. Zhang Q, Lu H, Kawazoe N, Chen G. Pore size effect of collagen scaffolds on cartilage regeneration. Acta Biomater. 2014;10:2005–13.

    CAS  Google Scholar 

  9. Huang BJ, Hu JC, Athanasiou KA. Cell-based tissue engineering strategies used in the clinical repair of articular cartilage. Biomaterials. 2016;98:1–22.

    CAS  Google Scholar 

  10. Yang J, Zhang YS, Yue K, Khademhosseini A. Cell-laden hydrogels for osteochondral and cartilage tissue engineering. Acta Biomater. 2017;57:1–25.

    CAS  Google Scholar 

  11. Ansboro S, Hayes J,S, Barron V, Browne S, Howard L, Greiser U et al. A chondromimetic microsphere for in situ spatially controlled chondrogenic differentiation of human mesenchymal stem cells. J Control Release. 2014;179:42–51.

    CAS  Google Scholar 

  12. Du M, Liang H, Mou C, Li X, Sun J, Zhuang Y et al. Regulation of human mesenchymal stem cells differentiation into chondrocytes in extracellular matrix-based hydrogel scaffolds. Colloids Surf B Biointerfaces. 2014;114:316–23.

    CAS  Google Scholar 

  13. Camarero-Espinosa S, Rothen-Rutishauser B, Foster EJ, Weder C. Articular cartilage: from formation to tissue engineering. Biomater Sci. 2016;4:734–67.

    CAS  Google Scholar 

  14. Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nat Publ Group. 2002;418:41–49.

    CAS  Google Scholar 

  15. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143–7.

    CAS  Google Scholar 

  16. Zuk PA, Zhu M, Ashjian P, De, Ugarte DA, Huang JI et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2014;13:4279–95.

    Google Scholar 

  17. Xue C, Shen Y, Li X, et al. Exosomes derived from hypoxia-treated human adipose mesenchymal stem cells enhance angiogenesis through the PKA signaling pathway. Stem Cells Dev. 2018;27:456–65.

    CAS  Google Scholar 

  18. Parmar PA, Chow LW, St-Pierre JP, Horejs CM, Peng YY, Werkmeister JA et al. Collagen-mimetic peptide-modifiable hydrogels for articular cartilage regeneration. Biomaterials. 2015;54:213–25.

    CAS  Google Scholar 

  19. Ravindran S, Kotecha M, Huang CC, Ye A, Pothirajan P, Yin Z et al. Biological and MRI characterization of biomimetic ECM scaffolds for cartilage tissue regeneration. Biomaterials. 2015;71:58–70.

    CAS  Google Scholar 

  20. Feng Q, Xu J, Zhang K, et al. Dynamic and cell-infiltratable hydrogels as injectable carrier of therapeutic cells and drugs for treating challenging bone defects. ACS Central Sci. 2019;5:440–50.

    CAS  Google Scholar 

  21. Iwasa J, Ochi M, Uchio Y, Katsube K, Adachi N, Kawasaki K. Effects of cell density on proliferation and matrix synthesis of chondrocytes embedded in atelocollagen gel. Artif Organs. 2003;27:249–55.

    CAS  Google Scholar 

  22. Takazawa K, Adachi N, Deie M, Kamei G, Uchio Y, Iwasa J et al. Evaluation of magnetic resonance imaging and clinical outcome after tissue-engineered cartilage implantation: prospective 6-year follow-up study. J Orthop Sci. 2012;17:413–24.

    Google Scholar 

  23. Zhang Y, Yu J, Ren K, et al. Thermosensitive hydrogels as scaffolds for cartilage tissue engineering. Bio Macromol. 2019;20:1478–92.

    CAS  Google Scholar 

  24. Ma Z, Gao C, Gong Y, Shen J. Cartilage tissue engineering PLLA scaffold with surface immobilized collagen and basic fibroblast growth factor. Biomaterials. 2005;26:1253–9.

    CAS  Google Scholar 

  25. Zhang K, Yan S, Li G, Cui L, Yin J. In-situ birth of MSCs multicellular spheroids in poly (L-glutamic acid)/chitosan scaffold for hyaline-like cartilage regeneration. Biomaterials. 2015;71:24–34.

    CAS  Google Scholar 

  26. Ren X, Li J, Li J, Jiang Y, Li L, Yao Q, et al. Aligned porous fibrous membrane with a biomimetic surface to accelerate cartilage regeneration. Chem Eng J. 2019;370:1027–38.

    CAS  Google Scholar 

  27. Grad S, Kupcsik L, Gorna K, Gogolewski S, Alini M. The use of biodegradable polyurethane scaffolds for cartilage tissue engineering: potential and limitations. Biomaterials. 2003;24:5163–71.

    CAS  Google Scholar 

  28. Meretoja VV, Dahlin RL, Wright S, Kasper FK, Mikos AG. Articular chondrocyte redifferentiation in 3D co-cultures with mesenchymal stem cells. Tissue Eng Part C. 2014;20:514–23.

    CAS  Google Scholar 

  29. Awad HA, Wickham MQ, Leddy HA, Gimble JM, Guilak F. Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. Biomaterials. 2004;25:3211–22.

    CAS  Google Scholar 

  30. Steinmetz NJ, Aisenbrey EA, Westbrook KK, Qi HJ, Bryant SJ. Mechanical loading regulates human MSC differentiation in a multi-layer hydrogel for osteochondral tissue engineering. Acta Biomaterialia. 2015;21:143–53.

    Google Scholar 

  31. Cui X, Breitenkamp K, Finn MG, Lotz M, D’Lima DD. Direct human cartilage repair using three-dimensional bioprinting technology. Tissue Eng Part A. 2012;18:1304–12.

    CAS  Google Scholar 

  32. Rosenzweig DH, Carelli E, Steffen T, Jarzem P, Haglund L. 3D-Printed ABS and PLA scaffolds for cartilage and nucleus pulposus tissue regeneration. Int J Mol Sci. 2015;16:15118–115135.

    CAS  Google Scholar 

  33. Yodmuang S, McNamara SL, Nover AB, Mandal BB, Agarwal M, Kelly TA et al. Silk microfiber-reinforced silk hydrogel composites for functional cartilage tissue repair. Acta Biomaterialia. 2015;11:27–36.

    CAS  Google Scholar 

  34. Lee CR, Breinan HA, Nehrer S, Spector M. Articular cartilage chondrocytes in type I and type II collagen-GAG matrices exhibit contractile behavior in vitro. Tissue Eng. 2000;6:555–65.

    CAS  Google Scholar 

  35. Meloni GR, Fisher MB, Stoeckl BD, Dodge GR, Mauck RL. Biphasic finite element modeling reconciles mechanical properties of tissue-engineered cartilage constructs across testing platforms. Tissue Eng—Part A. 2017;23:663–74.

    CAS  Google Scholar 

  36. Salimon AI, Statnik ES, Zadorozhnyy MY, et al. Porous open-cell UHMWPE: experimental study of structure and mechanical properties. Materials. 2019;12:2195.

    Google Scholar 

  37. Chen G, Ushida T, Tateishi T. A biodegradable hybrid sponge nested with collagen microsponges. J Biomed Mater Res. 2000;51:273–9.

    CAS  Google Scholar 

  38. Hu X, Zhu J, Li X, Zhang X, Meng Q, Yuan L et al. Dextran-coated fluorapatite crystals doped with Yb3+/Ho3+ for labeling and tracking chondrogenic differentiation of bone marrow mesenchymal stem cells in vitro and in vivo. Biomaterials. 2015;52:441–51.

    CAS  Google Scholar 

  39. Kim SH, Kim SH, Jung Y. TGF-β3 encapsulated PLCL scaffold by a supercritical CO2-HFIP co-solvent system for cartilage tissue engineering. J Control Release. 2015;206:101–7.

    CAS  Google Scholar 

  40. Wang Y, Kim UJ, Blasioli DJ, Kim HJ, Kaplan DL. In vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells. Biomaterials. 2015;26:7082–94.

    Google Scholar 

  41. Li D, Zou X-Y, El-Ayachi I, et al. Human dental pulp stem cells and gingival mesenchymal stem cells display action potential capacity in vitro after neuronogenic differentiation. Stem Cell Rev Rep. 2019;15:67–81.

    CAS  Google Scholar 

  42. Bertolo A, Baur M, Guerrero J, Pötzel T, Stoyanov J. Autofluorescence is a reliable in vitro marker of cellular senescence in human mesenchymal stromal cells. Sci Rep. 2019;9:2074.

    Google Scholar 

  43. Bonanomi A, Kojic D, Giger B, Rickenbach Z, Jean-Richard-Dit-Bressel L, Berger C et al. Quantitative cytokine gene expression in human tonsils at excision and during histoculture assessed by standardized and calibrated real-time PCR and novel data processing. J Immunol Methods. 2003;283:27–43.

    CAS  Google Scholar 

  44. Ding X, Zhu M, Xu B, Zhang J, Zhao Y, Ji S et al. Integrated trilayered silk fibroin scaffold for osteochondral differentiation of adipose-derived stem cells. Acs Appl Mater Interfaces. 2014;6:16696–705.

    CAS  Google Scholar 

  45. Wang X, Li Y, Han R, He C, Wang G, Wang J et al. Demineralized bone matrix combined bone marrow mesenchymal stem cells, bone morphogenetic protein-2 and transforming growth factor-β3 gene promoted pig cartilage defect repair. Plos ONE. 2014;9:e116061.

    Google Scholar 

  46. Xia W, Zhang L-L, Mo J, et al. Effect of static compression loads on intervertebral disc: an in vivo bent rat tail model. Orthop Surg. 2018;10:134–43.

    Google Scholar 

  47. Nakamuta. Y, Todo. M, Arahira T. Improvement of collagen gel/sponge composite scaffold by gel wrapping for cartilage. Tissue Eng Int J Biosci Biochem Bioinformatic. 2017;7:102–9. 2

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mitsugu Todo.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nakamuta, Y., Arahira, T. & Todo, M. Effects of culture conditions on the mechanical and biological properties of engineered cartilage constructed with collagen hybrid scaffold and human mesenchymal stem cells. J Mater Sci: Mater Med 30, 119 (2019). https://doi.org/10.1007/s10856-019-6321-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10856-019-6321-z

Navigation