Abstract

Research Article

Chondrogenic re-differentiation potential of chondrocytes after monolayer culture: Comparison between osteoarthritis and young adult patients

Kazuki Oishi*, Shusa Ohshika, Ken-Ichi Frukawa, Eiichi Tsuda, Yuji Yamamoto, Kazuki Oishi*, Shusa Ohshika, Ken-Ichi Frukawa, Eiichi Tsuda, Yuji Yamamoto and Yasuyuki Ishibashi

Published: 27 March, 2019 | Volume 4 - Issue 1 | Pages: 016-023

Purpose: Monolayer passage of chondrocytes results in dramatic phenotypic changes. This “de-differentiation” is expected to restore the chondrogenic properties such as “re-differentiation” in autologous chondrocyte implantation (ACI). The purpose of this study was to compare the chondrogenic re-differentiation potential of chondrocytes, from osteoarthritis (OA) patients and young adult patients, after monolayer culture.

Methods: Chondrocytes from five old patients with knee OA (OAC) and five young patients with recurrent shoulder dislocation (non-OAC) were used. The chondrocytes from passages 1 to 3 were analyzed for the expression of cell surface markers (CD73, CD90, CD105, and CD44) by flow cytometric analysis. Chondrocytes of passage 4 were cultured as pellets for re-differentiation and evaluated histologically. Real-time PCR were performed to measure the chondrogenic related genes transcriptional levels.

Results: OAC and non-OAC had comparable positive ratios for CD44, CD73, CD90, and CD105. The expression of CD105 was upregulated from passage 1 to passage 3 in OAC, and it increased at the same level as in non-OAC during passage 2 and 3. The expression of COL2 decreased from passage 1 to passage 3 in both the groups. There were no statistical differences in the Bern Scores between OAC and non-OAC.

Conclusion: The chondrocytes from OA patients and young adult patients had chondrogenic re-differentiation potential. The changes in cell surface markers and chondrogenic related genes showed similarity for both the groups. Our findings suggest that OAC can become the cell source for ACI.

Read Full Article HTML DOI: 10.29328/journal.jsmt.1001038 Cite this Article Read Full Article PDF

References

  1. Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, et al. Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation, N Engl J Med. 1994; 331: 889-895. Ref.: https://goo.gl/hRWS2M
  2. Goyal D, Goyal A, Keyhani S, Lee EH, Hui JH Evidence-based status of second- and third-generation autologous chondrocyte implantation over first generation: a systematic review of level I and II studies. Arthroscopy. 2013; 29: 1872-1878. Ref.: https://goo.gl/9H4U1r
  3. Oussedik S, Tsitskaris K, Parker D. Treatment of articular cartilage lesions of the knee by microfracture or autologous chondrocyte implantation: a systematic review. Arthroscopy. 2015; 4: 732-744. Ref.: https://goo.gl/wdQkML
  4. Benya PD, Shaffer JD. Shaffer. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 1982; 30: 215-224. Ref.: https://goo.gl/6h3gTg
  5. Diaz-Romero J, Gaillard JP, Grogan SP, Nesic D, Trub T, et al. Immunophenotypic analysis of human articular chondrocytes: changes in surface markers associated with cell expansion in monolayer culture. J Cell Physiol. 2005; 202: 731-742. Ref.: https://goo.gl/BxJxsC
  6. Bernstein P, Sperling I, Corbeil D, Hempel U, Fickert S Progenitor cells from cartilage--no osteoarthritis-grade-specific differences in stem cell marker expression. Biotechnol Prog. 2013; 29: 206-212. Ref.: https://goo.gl/TiYbEz
  7. Salamon A, Jonitz-Heincke A, Adam S, Rychly J, Müller-Hilke B, et al. Articular cartilage-derived cells hold a strong osteogenic differentiation potential in comparison to mesenchymal stem cells in vitro. Exp Cell Res. 2013; 319: 2856-2865. Ref.: https://goo.gl/wR4LQn
  8. Yoshimura N, Muraki S, Oka H, Mabuchi A, En-Yo Y, et al. Prevalence of knee osteoarthritis, lumbar spondylosis, and osteoporosis in Japanese men and women: the research on osteoarthritis/osteoporosis against disability study. J Bone Miner Metab. 2009; 27: 620-628. Ref.: https://goo.gl/BxZLk1
  9. Yang KG, Saris DB, Geuze RE, van Rijen MH, van der Helm YJ, et al. Altered in vitro chondrogenic properties of chondrocytes harvested from unaffected cartilage in osteoarthritic joints. Osteoarthr Cartil. 2006; 14: 561-570. Ref.: https://goo.gl/qWSF5M
  10. Dehne T, Karlsson C, Ringe J, Sittinger M, Lindahl A. Chondrogenic differentiation potential of osteoarthritic chondrocytes and their possible use in matrix-associated autologous chondrocyte transplantation. Arthritis Res Ther. 2009; 11: R133. Ref.: https://goo.gl/SCnRKp
  11. Lin Z, Fitzgerald JB, Xu J, Willers C, Wood D, et al. Gene expression profiles of human chondrocytes during passaged monolayer cultivation. J Orthop Res. 2008; 26: 1230-1237. Ref.: https://goo.gl/LHW7g8
  12. Tanaka N, Ikeda Y, Yamaguchi T, Furukawa H, Mitomi H, et al. α5β1 integrin induces the expression of noncartilaginous procollagen gene expression in articular chondrocytes cultured in monolayers.. Arthritis Res Ther. 2013; 15: R127. Ref.: https://goo.gl/5t19th
  13. Aulthouse AL, Beck M, Griffey E, Sanford J, Arden K, et al. Expression of the human chondrocyte phenotype in vitro. In Vitro Cell Dev Biol. 1989; 7: 659-668. Ref.: https://goo.gl/bEFGVp
  14. Grogan SP, Barbero A, Winkelmann V, Rieser F, Fitzsimmons JS, et al. Visual histological grading system for the evaluation of in vitro-generated neocartilage. Tissue Eng. 2006; 12: 2141-2149. Ref.: https://goo.gl/BVChBA
  15. Pierelli L, Bonanno G, Rutella S, Marone M, Scambia G, et al. CD105 (endoglin) expression on hematopoietic stem/progenitor cells. Leuk Lymphoma. 2001; 42: 1195-1206. Ref.: https://goo.gl/k96nwV
  16. Jaggupilli A, Elkord E. Significance of CD44 and CD24 as cancer stem cell markers: an enduring ambiguity. Clin Dev Immunol. 2012; 2012: 708036. Ref.: https://goo.gl/ysTb1K
  17. Jonitz A, Lochner K, Peters K, Salamon A, Pasold J, et al. Differentiation capacity of human chondrocytes embedded in alginate matrix. Connect Tissue Res. 2011; 52: 503-511. Ref.: https://goo.gl/9bJZwJ
  18. Schnabel M1, Marlovits S, Eckhoff G, Fichtel I, Gotzen L, et al. Dedifferentiation-associated changes in morphology and gene expression in primary human articular chondrocytes in cell culture. Osteoarthr Cartil. 2002; 10: 62-70. Ref.: https://goo.gl/SJNXeA
  19. von der Mark K, Gauss V, von der Mark H, Müller P. Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature. 1977; 267: 531-532. Ref.: https://goo.gl/oJe9xE
  20. Novakofski KD, Berg LC, Bronzini I, Bonnevie ED, Poland SG, et al. Joint-dependent response to impact and implications for post-traumatic osteoarthritis, Osteoarthr. Cartil. 2015; 23: 1130-1137. Ref.: https://goo.gl/CDXqEf
  21. Vincenti MP, Brinckerhoff CE. Brinckerhoff. Transcriptional regulation of collagenase (MMP-1, MMP-13) genes in arthritis: integration of complex signaling pathways for the recruitment of gene-specific transcription factors. Arthritis Res. 2002; 4: 157-164. Ref.: https://goo.gl/aGzdjA
  22. Barbero A, Grogan S, Schäfer D, Heberer M, Mainil-Varlet P, et al. Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity. Osteoarthr Cartil. 2004; 12: 476-484. Ref.: https://goo.gl/fHfDkd

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