Skip to main content
Log in

Cartilage tissue engineering using chondrocyte-derived extracellular matrix scaffold suppressed vessel invasion during chondrogenesis of mesenchymal stem cells in vivo

  • Original Article
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Loss of chondrogenic phenotypes of tissue engineered cartilage using mesenchymal stem cells (MSCs) in vivo are thought to be influenced by environmental factors like vessel invasion in particular. This study investigated effect of a chondrocyte-derived extracellular matrix (CD-ECM) scaffold on the hypertrophic changes and vessel invasion into tissue engineered cartilage using rabbit MSCs in comparison with a synthetic polyglycolic acid (PGA) scaffold. Rabbit MSCs in CD-ECM or PGA scaffold were differentiated for 1 week in vitro and implanted in the back of nude mice for 6 weeks in vivo. Gross observation showed red stains, indicative of vessel invasion, increased along with the loss of chondrogenic phenotype in safranin-O stains, which was more prominent in the PGA constructs. The area showing loss of chondrogenic phenotypes in safranin-O stain was correlated well with the mineralized area in the von kossa stain and the area with vessel-like structures in the gomori aldehyde fuchsin stain at 6 weeks in terms of their size and distribution. Also, vessel invasion took place more deeply and intensively into the constructs, in accordance with the expression of angiogenic markers (CD31, VEGF-A and HIF-1α) and a macrophage marker (CD68). This phenomenon progressed much more rapidly in the PGA constructs than in the CDECM constructs, and correlated well with the loss of chondrogenic phenotypes. In conclusion, this study showed that tissue engineered cartilage using the CD-ECM scaffold maintained better chondrogenic phenotypes in vivo and showed lower levels of hypertrophic changes and vessel invasion.

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

Similar content being viewed by others

References

  1. A Dickhut, K Pelttari, P Janicki, et al., Calcification or dedifferentiation: requirement to lock mesenchymal stem cells in a desired differentiation stage, J cell physiol, 219, 219 (2009).

    Article  PubMed  CAS  Google Scholar 

  2. JH Cui, SR Park, K Park, et al., Preconditioning of mesenchymal stem cells with low-intensity ultrasound for cartilage formation in vivo, Tissue eng, 13, 351 (2007).

    Article  PubMed  CAS  Google Scholar 

  3. K Pelttari, A Winter, E Steck, et al., 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, 54, 3254 (2006).

    Article  PubMed  CAS  Google Scholar 

  4. S Ichinose, K Yamagata, I Sekiya, et al., Detailed examination of cartilage formation and endochondral ossification using human mesenchymal stem cells, Clin Exp pharmacol physiol, 32, 561 (2005).

    Article  PubMed  CAS  Google Scholar 

  5. RS Tuan, Stemming cartilage degeneration: adult mesenchymal stem cells as a cell source for articular cartilage tissue engineering, Arthritis Rheum, 54, 3075 (2006).

    Article  PubMed  CAS  Google Scholar 

  6. M Murata, K Yudoh, K Masuko, The potential role of vascular endothelial growth factor (VEGF) in cartilage How the angiogenic factor could be involved in the pathogenesis of osteoarthritis?, Osteoarthritis and cartilage, 16, 279 (2008).

    Article  PubMed  CAS  Google Scholar 

  7. S Ashraf, DA Walsh, Angiogenesis in osteoarthritis, Curr opin rheumatol, 20, 573 (2008).

    Article  PubMed  Google Scholar 

  8. CS Adams, IM Shapiro, The fate of the terminally differentiated chondrocyte: evidence for microenvironmental regulation of chondrocyte apoptosis, Crit Rev Oral Biol Med, 13, 465 (2002).

    Article  PubMed  Google Scholar 

  9. R Cancedda, P Castagnola, FD Cancedda, et al., Developmental control of chondrogenesis and osteogenesis, Int J Dev Biol, 44, 707 (2000).

    PubMed  CAS  Google Scholar 

  10. C Yoshioka, T Yagi, Electron microscopic observations on the fate of hypertrophic chondrocytes in condylar cartilage of rat mandible, J craniofac genetics dev biol, 8, 253 (1988).

    CAS  Google Scholar 

  11. CS Adams, IM Shapiro, The fate of the terminally differentiated chondrocyte: evidence for microenvironmental regulation of chondrocyte apoptosis, Crit Rev Oral Biol Med, 13, 465 (2002).

    Article  PubMed  Google Scholar 

  12. C De Bari, F Dell’Accio, FP Luyten, Failure of in vitro-differentiated mesenchymal stem cells from the synovial membrane to form ectopic stable cartilage in vivo, Arthritis Rheum, 50, 142 (2004).

    Article  PubMed  Google Scholar 

  13. F Dell’Accio, CD Bari, FP Luyten, Microenvironment and phenotypic stability specify tissue formation by human articular cartilage-derived cells in vivo, Exp cell res, 287, 16 (2003).

    Article  PubMed  Google Scholar 

  14. R Stoop, Smart biomaterials for tissue engineering of cartilage, Injury, 39, 77 (2008).

    Article  Google Scholar 

  15. R Seda Tigli, S Ghosh, MM Laha, et al., Comparative chondrogenesis of human cell sources in 3D scaffolds, J tissue eng and regene med, 3, 348 (2009).

    Article  CAS  Google Scholar 

  16. JL Puetzer, JN Petitte, EG Loboa, Comparative review of growth factors for induction of three-dimensional in vitro chondrogenesis in human mesenchymal stem cells isolated from bone marrow and adipose tissue, tissue Eng Part B: Reviews, 16, 435 (2010).

    Article  CAS  Google Scholar 

  17. X Yang, L Chen, X Xu, et al., TGF-β/Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage, J cell biol, 153, 35 (2001).

    Article  PubMed  CAS  Google Scholar 

  18. P Allen, J Melero-Martin, J Bischoff, Type I collagen, fibrin and puramatrix matrices provide permissive environments for human endothelial and mesenchymal progenitor cells to form neovascular networks, J tissue eng regen med, 5, 74 (2011).

    Article  Google Scholar 

  19. C Sosio, F Boschetti, L Mangiavini, et al., Blood exposure has a negative effect on engineered cartilage, Knee Surg, Sports Traumatol, Arthrosc, 19, 4035–1 (2010).

    Google Scholar 

  20. CZ Jin, SR Park, BH Choi, et al., In vivo cartilage tissue engineering using a cell derived extracellular matrix scaffold, Artifi Organs, 31, 183 (2007).

    Article  CAS  Google Scholar 

  21. CZ Jin, BH Choi, SR Park, et al., Cartilage engineering using cell-derived extracellular matrix scaffold in vitro, J Biomed Mater Res Part A, 92, 1567 (2010).

    Google Scholar 

  22. KH Choi, BH Choi, SR Park, et al., The chondrogenic differentiation of mesenchymal stem cells on an extracellular matrix scaffold derived from porcine chondrocytes, Biomaterials, (2010).

  23. TJ Collins, ImageJ for microscopy, Biotechniques, 43, 25 (2007).

    Article  PubMed  Google Scholar 

  24. M Alves da Silva, A Crawford, J Mundy, et al., Chitosan/polyester-based scaffolds for cartilage tissue engineering: assessment of extracellular matrix formation, Acta Biomater, 6, 1149 (2010).

    Article  PubMed  CAS  Google Scholar 

  25. R Brown, C McFarland, Regulation of growth plate cartilage degradation in vitro: effects of calcification and a low molecular weight angiogenic factor (ESAF)* 1, Bone miner, 17, 49 (1992).

    Article  PubMed  CAS  Google Scholar 

  26. CF Descalzi, A Melchiori, R Benelli, et al., Production of angiogenesis inhibitors and stimulators is modulated by cultured growth plate chondrocytes during in vitro differentiation: dependence on extracellular matrix assembly, Eur j cell biol, 66, 60 (1995).

    Google Scholar 

  27. HP Gerber, TH Vu, AM Ryan, et al., VEGF couples hypertrophic cartilage remodeling, ossification and angiogenesis during endochondral bone formation, Nat med, 5, 623 (1999).

    Article  PubMed  CAS  Google Scholar 

  28. HH Kim, SE Lee, WJ Chung, et al., Stabilization of hypoxia-inducible factor-1 [alpha] is involved in the hypoxic stimuli-induced expression of vascular endothelial growth factor in osteoblastic cells, Cytokine, 17, 14 (2002).

    Article  PubMed  CAS  Google Scholar 

  29. DE Dobson, A Kambe, E Block, et al., 1-Butyryl-glycerol: a novel angiogenesis factor secreted by differentiating adipocytes, Cell, 61, 223 (1990).

    Article  PubMed  CAS  Google Scholar 

  30. F Yi, Yi W Pin, Xu Z Dong, et al., Endostatin promotes the anabolic program of rabbit chondrocyte, Cell Res, 15, 201 (2005).

    Article  Google Scholar 

  31. JR Jackson, M Seed, C Kircher, et al., The codependence of angiogenesis and chronic inflammation, FASEB j, 11, 457 (1997).

    PubMed  CAS  Google Scholar 

  32. MML Deckers, ER Van Beek, G Van Der Pluijm, et al., Dissociation of angiogenesis and osteoclastogenesis during endochondral bone formation in neonatal mice, J Bone Min Res, 17, 998 (2002).

    Article  Google Scholar 

  33. JE Valentin, AM Stewart-Akers, TW Gilbert, et al., Macrophage participation in the degradation and remodeling of extracellular matrix scaffolds, Tissue Eng Part A, 15, 1687 (2009).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Byoung-Hyun Min.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Choi, KH., Song, B.R., Choi, B.H. et al. Cartilage tissue engineering using chondrocyte-derived extracellular matrix scaffold suppressed vessel invasion during chondrogenesis of mesenchymal stem cells in vivo . Tissue Eng Regen Med 9, 43–50 (2012). https://doi.org/10.1007/s13770-012-0043-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13770-012-0043-3

Key words

Navigation