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Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture

Year 2022, Volume 15, Issue 2, 217 - 222, 30.06.2022
https://doi.org/10.30607/kvj.1081105

Abstract

Mesenchymal stem cells are widely used in the treatment of many diseases, including osteoarthritis, due to their ability to differentiate into cartilage. The high chondrogenic differentiation potential of synovial fluid-derived mesenchymal stem cells increases the importance of these cells in osteoarthritis treatments. Addition of BMP-9 and TGF-ß3 into chondrogenic differentiation medium, increases chondrogenic differentiation and they also cause hypertrophic effects on chondrocytes. In our study, it was aimed to demonstrate the effects of BMP-9 and TGF-ß3 on cell hypertrophy by adding them into the chondrogenic basal medium during in vitro chondrogenic differentiation. In the study, stem cells in passage 5 and chondrocytes in passage 1 were cultured in a transwell co-culture system and six experimental groups were formed. Cell hypertrophy was demonstrated by examining MMP-13 and RUNX-2 gene expressions, in stem cells where chondrogenesis were induced in transwell co-culture. Although the addition of BMP-9 and TGF-ß3 to the chondrogenic medium increased hypertrophic gene expressions in experimental groups compared to control, the results were not statistically significant. The addition of BMP-9 and TGF-ß3, separately or in combination, during the chondrogenic differentiation of stem cells does not cause significant chondrocyte hypertrophy.

References

  • Barry F, Boynton RE, Liu B, Murphy JM. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res. 2001 Aug 15;268(2):189-200.
  • Blunk T, Sieminski AL, Appel B, Croft C, Courter DL, Chieh JJ, Goepferich A, Khurana JS, Gooch KJ. Bone morphogenetic protein 9: a potent modulator of cartilage development in vitro. Growth Factors. 2003 Jun;21(2):71-7.
  • Cheng A, Gustafson AR, Schaner Tooley CE, Zhang M. BMP-9 dependent pathways required for the chondrogenic differentiation of pluripotent stem cells. Differentiation. 2016 Dec;92(5):298-305. doi: 10.1016/j.diff.2016.03.005. Epub 2016 Apr 4.
  • Derfoul A, Perkins GL, Hall DJ, Tuan RS. Glucocorticoids promote chondrogenic differentiation of adult human mesenchymal stem cells by enhancing expression of cartilage extracellular matrix genes. Stem Cells. 2006 Jun;24(6):1487-95.
  • Deng ZH, Li YS, Gao X, Lei GH, Huard J. Bone morphogenetic proteins for articular cartilage regeneration. Osteoarthritis Cartilage. 2018 Sep;26(9):1153-1161.
  • Jones EA, Crawford A, English A, Henshaw K, Mundy J, Corscadden D, Chapman T, Emery P, Hatton P, McGonagle D. Synovial fluid mesenchymal stem cells in health and early osteoarthritis: detection and functional evaluation at the single-cell level. Arthritis Rheum. 2008 Jun;58(6):1731-40.
  • Kubosch EJ, Heidt E, Bernstein A, Böttiger K, Schmal H. The trans-well coculture of human synovial mesenchymal stem cells with chondrocytes leads to self-organization, chondrogenic differentiation, and secretion of TGFβ. Stem Cell Res Ther. 2016 Apr 26;7(1):64.
  • Kurth T, Hedbom E, Shintani N, Sugimoto M, Chen FH, Haspl M, Martinovic S, Hunziker EB. Chondrogenic potential of human synovial mesenchymal stem cells in alginate. Osteoarthritis Cartilage. 2007 Oct;15(10):1178-89. doi: 10.1016/j.joca.2007.03.015. Epub 2007 May 14.
  • Majumdar MK, Wang E, Morris EA. BMP-2 and BMP-9 promotes chondrogenic differentiation of human multipotential mesenchymal cells and overcomes the inhibitory effect of IL-1. J Cell Physiol. 2001 Dec;189(3):275-84.
  • Massagué J, Wotton D. Transcriptional control by the TGF-beta/Smad signaling system. EMBO J. 2000 Apr 17;19(8):1745-54.
  • Morgan BJ, Bauza-Mayol G, Gardner OFW, Zhang Y, Levato R, Archer CW, van Weeren R, Malda J, Conlan RS, Francis LW, Khan IM. Bone Morphogenetic Protein-9 Is a Potent Chondrogenic and Morphogenic Factor for Articular Cartilage Chondroprogenitors. Stem Cells Dev. 2020 Jul;29(14):882-894.
  • Murata D, Miyakoshi D, Hatazoe T, Miura N, Tokunaga S, Fujiki M, Nakayama K, Misumi K. Multipotency of equine mesenchymal stem cells derived from synovial fluid. Vet J. 2014 Oct;202(1):53-61.
  • Qing C, Wei-ding C, Wei-min F. Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components. Braz J Med Biol Res. 2011 Apr;44(4):303-10.
  • Ren X, Weisgerber DW, Bischoff D, Lewis MS, Reid RR, He TC, Yamaguchi DT, Miller TA, Harley BA, Lee JC. Nanoparticulate Mineralized Collagen Scaffolds and BMP-9 Induce a Long-Term Bone Cartilage Construct in Human Mesenchymal Stem Cells. Adv Healthc Mater. 2016 Jul;5(14):1821-30.
  • Scarfì S. Use of bone morphogenetic proteins in mesenchymal stem cell stimulation of cartilage and bone repair. World J Stem Cells. 2016 Jan 26;8(1):1-12.
  • Sekiya I, Katano H, Ozeki N. Characteristics of MSCs in Synovial Fluid and Mode of Action of Intra-Articular Injections of Synovial MSCs in Knee Osteoarthritis. Int J Mol Sci. 2021 Mar 11;22(6):2838
  • Tang QO, Shakib K, Heliotis M, Tsiridis E, Mantalaris A, Ripamonti U, Tsiridis E. TGF-beta3: A potential biological therapy for enhancing chondrogenesis. Expert Opin Biol Ther. 2009 Jun;9(6):689-701.
  • van Caam A, Blaney Davidson E, Garcia de Vinuesa A, van Geffen E, van den Berg W, Goumans MJ, ten Dijke P, van der Kraan P. The high affinity ALK1-ligand BMP9 induces a hypertrophy-like state in chondrocytes that is antagonized by TGFβ1. Osteoarthritis Cartilage. 2015 Jun;23(6):985-95.
  • van der Kraan PM, Blaney Davidson EN, Blom A, van den Berg WB. TGF-beta signaling in chondrocyte terminal differentiation and osteoarthritis: modulation and integration of signaling pathways through receptor-Smads. Osteoarthritis Cartilage. 2009 Dec;17(12):1539-45.
  • Xu L, Wu Y, Liu Y, Zhou Y, Ye Z, Tan WS. Non-contact Coculture Reveals a Comprehensive Response of Chondrocytes Induced by Mesenchymal Stem Cells Through Trophic Secretion. Tissue Eng Regen Med. 2017 Sep 27;15(1):37-48.
  • Wang X, Manner PA, Horner A, Shum L, Tuan RS, Nuckolls GH. Regulation of MMP-13 expression by RUNX2 and FGF2 in osteoarthritic cartilage. Osteoarthritis Cartilage. 2004 Dec;12(12):963-73.
  • Wu L, Prins HJ, Helder MN, van Blitterswijk CA, Karperien M. Trophic effects of mesenchymal stem cells in chondrocyte co-cultures are independent of culture conditions and cell sources. Tissue Eng Part A. 2012 Aug;18(15-16):1542-51.

BMP-9 ve TGF-ß3 Eklenmiş Kondrojenik Farklılaşma Medyumunun Transwell Ko-kültürde Hipertrofi Üzerine Etkisi

Year 2022, Volume 15, Issue 2, 217 - 222, 30.06.2022
https://doi.org/10.30607/kvj.1081105

Abstract

Mezenkimal kök hücreler, kıkırdağa farklılaşma yetenekleri nedeniyle osteoartrit dahil birçok hastalığın tedavisinde yaygın olarak kullanılmaktadır. Sinoviyal sıvı kökenli mezenkimal kök hücrelerinin kondrojenik farklılaşma potansiyellerinin fazla olması bu hücrelerin osteoartrit tedavilerindeki önemini artırmaktadır. Kondrojenik farklılaşma medyumuna ilave edilen BMP-9 ve TGF-ß3 büyüme faktörleri kondrojenik farklılaşmayı artırır ve aynı zamanda kondrositlerde hipertofik etkilere sebep olur. Çalışmamızda in vitro kondrojenik farklılaşma esnasında kondrojenik bazal medyuma ilave edilen BMP-9 ve TGF-ß3’ün hücre hipertrofisi üzerine etkilerinin gösterilmesi amaçlandı. Çalışmada transwell ko-kültür sisteminde 5. pasajdaki kök hücreler ve pasaj 1'deki kondrositler birlikte kültüre edildi ve altı deney grubu oluşturuldu. Hücre hipertrofisi, transwell ko-kültüründe kondrogenezin indüklendiği kök hücrelerde MMP-13 ve RUNX-2 gen ekspresyonları incelenerek gösterildi. BMP-9 ve TGF-ß3'ün kondrojenik ortama eklenmesi, kontrol grubuna göre deney gruplarında hipertrofik gen ekspresyonlarını artırmasına rağmen bu artış istatistiksel olarak anlamlı değildi. Kök hücrelerin kondrojenik farklılaşması esnasında BMP-9 ve TGF-ß3’ün ayrı ayrı veya birlikte kullanılması önemli derecede kıkırdak hücresi hipertrofisine neden olmamaktadır. 

References

  • Barry F, Boynton RE, Liu B, Murphy JM. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res. 2001 Aug 15;268(2):189-200.
  • Blunk T, Sieminski AL, Appel B, Croft C, Courter DL, Chieh JJ, Goepferich A, Khurana JS, Gooch KJ. Bone morphogenetic protein 9: a potent modulator of cartilage development in vitro. Growth Factors. 2003 Jun;21(2):71-7.
  • Cheng A, Gustafson AR, Schaner Tooley CE, Zhang M. BMP-9 dependent pathways required for the chondrogenic differentiation of pluripotent stem cells. Differentiation. 2016 Dec;92(5):298-305. doi: 10.1016/j.diff.2016.03.005. Epub 2016 Apr 4.
  • Derfoul A, Perkins GL, Hall DJ, Tuan RS. Glucocorticoids promote chondrogenic differentiation of adult human mesenchymal stem cells by enhancing expression of cartilage extracellular matrix genes. Stem Cells. 2006 Jun;24(6):1487-95.
  • Deng ZH, Li YS, Gao X, Lei GH, Huard J. Bone morphogenetic proteins for articular cartilage regeneration. Osteoarthritis Cartilage. 2018 Sep;26(9):1153-1161.
  • Jones EA, Crawford A, English A, Henshaw K, Mundy J, Corscadden D, Chapman T, Emery P, Hatton P, McGonagle D. Synovial fluid mesenchymal stem cells in health and early osteoarthritis: detection and functional evaluation at the single-cell level. Arthritis Rheum. 2008 Jun;58(6):1731-40.
  • Kubosch EJ, Heidt E, Bernstein A, Böttiger K, Schmal H. The trans-well coculture of human synovial mesenchymal stem cells with chondrocytes leads to self-organization, chondrogenic differentiation, and secretion of TGFβ. Stem Cell Res Ther. 2016 Apr 26;7(1):64.
  • Kurth T, Hedbom E, Shintani N, Sugimoto M, Chen FH, Haspl M, Martinovic S, Hunziker EB. Chondrogenic potential of human synovial mesenchymal stem cells in alginate. Osteoarthritis Cartilage. 2007 Oct;15(10):1178-89. doi: 10.1016/j.joca.2007.03.015. Epub 2007 May 14.
  • Majumdar MK, Wang E, Morris EA. BMP-2 and BMP-9 promotes chondrogenic differentiation of human multipotential mesenchymal cells and overcomes the inhibitory effect of IL-1. J Cell Physiol. 2001 Dec;189(3):275-84.
  • Massagué J, Wotton D. Transcriptional control by the TGF-beta/Smad signaling system. EMBO J. 2000 Apr 17;19(8):1745-54.
  • Morgan BJ, Bauza-Mayol G, Gardner OFW, Zhang Y, Levato R, Archer CW, van Weeren R, Malda J, Conlan RS, Francis LW, Khan IM. Bone Morphogenetic Protein-9 Is a Potent Chondrogenic and Morphogenic Factor for Articular Cartilage Chondroprogenitors. Stem Cells Dev. 2020 Jul;29(14):882-894.
  • Murata D, Miyakoshi D, Hatazoe T, Miura N, Tokunaga S, Fujiki M, Nakayama K, Misumi K. Multipotency of equine mesenchymal stem cells derived from synovial fluid. Vet J. 2014 Oct;202(1):53-61.
  • Qing C, Wei-ding C, Wei-min F. Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components. Braz J Med Biol Res. 2011 Apr;44(4):303-10.
  • Ren X, Weisgerber DW, Bischoff D, Lewis MS, Reid RR, He TC, Yamaguchi DT, Miller TA, Harley BA, Lee JC. Nanoparticulate Mineralized Collagen Scaffolds and BMP-9 Induce a Long-Term Bone Cartilage Construct in Human Mesenchymal Stem Cells. Adv Healthc Mater. 2016 Jul;5(14):1821-30.
  • Scarfì S. Use of bone morphogenetic proteins in mesenchymal stem cell stimulation of cartilage and bone repair. World J Stem Cells. 2016 Jan 26;8(1):1-12.
  • Sekiya I, Katano H, Ozeki N. Characteristics of MSCs in Synovial Fluid and Mode of Action of Intra-Articular Injections of Synovial MSCs in Knee Osteoarthritis. Int J Mol Sci. 2021 Mar 11;22(6):2838
  • Tang QO, Shakib K, Heliotis M, Tsiridis E, Mantalaris A, Ripamonti U, Tsiridis E. TGF-beta3: A potential biological therapy for enhancing chondrogenesis. Expert Opin Biol Ther. 2009 Jun;9(6):689-701.
  • van Caam A, Blaney Davidson E, Garcia de Vinuesa A, van Geffen E, van den Berg W, Goumans MJ, ten Dijke P, van der Kraan P. The high affinity ALK1-ligand BMP9 induces a hypertrophy-like state in chondrocytes that is antagonized by TGFβ1. Osteoarthritis Cartilage. 2015 Jun;23(6):985-95.
  • van der Kraan PM, Blaney Davidson EN, Blom A, van den Berg WB. TGF-beta signaling in chondrocyte terminal differentiation and osteoarthritis: modulation and integration of signaling pathways through receptor-Smads. Osteoarthritis Cartilage. 2009 Dec;17(12):1539-45.
  • Xu L, Wu Y, Liu Y, Zhou Y, Ye Z, Tan WS. Non-contact Coculture Reveals a Comprehensive Response of Chondrocytes Induced by Mesenchymal Stem Cells Through Trophic Secretion. Tissue Eng Regen Med. 2017 Sep 27;15(1):37-48.
  • Wang X, Manner PA, Horner A, Shum L, Tuan RS, Nuckolls GH. Regulation of MMP-13 expression by RUNX2 and FGF2 in osteoarthritic cartilage. Osteoarthritis Cartilage. 2004 Dec;12(12):963-73.
  • Wu L, Prins HJ, Helder MN, van Blitterswijk CA, Karperien M. Trophic effects of mesenchymal stem cells in chondrocyte co-cultures are independent of culture conditions and cell sources. Tissue Eng Part A. 2012 Aug;18(15-16):1542-51.

Details

Primary Language English
Subjects Veterinary
Journal Section RESEARCH ARTICLE
Authors

Elif Ece AKGÜN> (Primary Author)
Afyon Kocatepe University, Veterinary Faculty, Histology and Embryology Department
0000-0002-2982-0307
Türkiye


Eda DEMİRTAŞ>
Afyon Kocatepe University, Veterinary Faculty, Genetics Department
0000-0002-3061-0091
Türkiye


Özlem ÖZDEN AKKAYA>
Afyon Kocatepe University, Veterinary Faculty, Histology and Embryology Department
0000-0001-6372-9155
Türkiye


Metin ERDOĞAN>
Afyon Kocatepe University, Veterinary Faculty, Genetics Department
0000-0003-0975-1989
Türkiye


Korhan ALTUNBAŞ>
Afyon Kocatepe University, Veterinary Faculty, Histology and Embryology Department
0000-0001-7580-3412
Türkiye

Publication Date June 30, 2022
Published in Issue Year 2022, Volume 15, Issue 2

Cite

Bibtex @research article { kvj1081105, journal = {Kocatepe Veterinary Journal}, issn = {1308-1594}, eissn = {2147-6853}, address = {}, publisher = {Afyon Kocatepe University}, year = {2022}, volume = {15}, number = {2}, pages = {217 - 222}, doi = {10.30607/kvj.1081105}, title = {Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture}, key = {cite}, author = {Akgün, Elif Ece and Demirtaş, Eda and Özden Akkaya, Özlem and Erdoğan, Metin and Altunbaş, Korhan} }
APA Akgün, E. E. , Demirtaş, E. , Özden Akkaya, Ö. , Erdoğan, M. & Altunbaş, K. (2022). Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture . Kocatepe Veterinary Journal , 15 (2) , 217-222 . DOI: 10.30607/kvj.1081105
MLA Akgün, E. E. , Demirtaş, E. , Özden Akkaya, Ö. , Erdoğan, M. , Altunbaş, K. "Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture" . Kocatepe Veterinary Journal 15 (2022 ): 217-222 <https://dergipark.org.tr/en/pub/kvj/issue/69281/1081105>
Chicago Akgün, E. E. , Demirtaş, E. , Özden Akkaya, Ö. , Erdoğan, M. , Altunbaş, K. "Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture". Kocatepe Veterinary Journal 15 (2022 ): 217-222
RIS TY - JOUR T1 - Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture AU - Elif Ece Akgün , Eda Demirtaş , Özlem Özden Akkaya , Metin Erdoğan , Korhan Altunbaş Y1 - 2022 PY - 2022 N1 - doi: 10.30607/kvj.1081105 DO - 10.30607/kvj.1081105 T2 - Kocatepe Veterinary Journal JF - Journal JO - JOR SP - 217 EP - 222 VL - 15 IS - 2 SN - 1308-1594-2147-6853 M3 - doi: 10.30607/kvj.1081105 UR - https://doi.org/10.30607/kvj.1081105 Y2 - 2022 ER -
EndNote %0 Kocatepe Veterinary Journal Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture %A Elif Ece Akgün , Eda Demirtaş , Özlem Özden Akkaya , Metin Erdoğan , Korhan Altunbaş %T Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture %D 2022 %J Kocatepe Veterinary Journal %P 1308-1594-2147-6853 %V 15 %N 2 %R doi: 10.30607/kvj.1081105 %U 10.30607/kvj.1081105
ISNAD Akgün, Elif Ece , Demirtaş, Eda , Özden Akkaya, Özlem , Erdoğan, Metin , Altunbaş, Korhan . "Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture". Kocatepe Veterinary Journal 15 / 2 (June 2022): 217-222 . https://doi.org/10.30607/kvj.1081105
AMA Akgün E. E. , Demirtaş E. , Özden Akkaya Ö. , Erdoğan M. , Altunbaş K. Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture. Kocatepe Veterinary Journal. 2022; 15(2): 217-222.
Vancouver Akgün E. E. , Demirtaş E. , Özden Akkaya Ö. , Erdoğan M. , Altunbaş K. Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture. Kocatepe Veterinary Journal. 2022; 15(2): 217-222.
IEEE E. E. Akgün , E. Demirtaş , Ö. Özden Akkaya , M. Erdoğan and K. Altunbaş , "Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture", Kocatepe Veterinary Journal, vol. 15, no. 2, pp. 217-222, Jun. 2022, doi:10.30607/kvj.1081105

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