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The evaluation of cartilage differentiations using transforming growth factor beta3 alone and with combination of bone morphogenetic protein-6 on adult stem cells

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Abstract

In our quest to standardize our formula for a clinical trial, transforming growth factor-beta3 (TGF-β3) alone and in combination with bone morphogenetic protein-6 (BMP-6) were evaluated for their effectiveness in cartilage differentiation. Bone Marrow Stem Cells (BMSCs) and Adipose Derived Stem Cells (ADSCs) were induced to chondrogenic lineage using two different media. Native chondrocytes served as positive control. ADSCs and BMSCs proved multipotency by tri-lineage differentiations. ADSC has significantly higher growth kinetics compare to Chondrocyte only p ≤ 0.05. Using TGF-β3 alone, BMSC revealed higher expressions for hyaline cartilage genes compare to ADSCs. Chondrocyte has significantly higher early chondrogenic markers expression to ADSCs and BMSCs, while BMSCs was only higher to ADSC at chondroadherin, p ≤ 0.0001. On mature chondrogenic markers, chondrocytes were significantly higher to ADSCs and BMSCs for aggrecan, collagen IX, sry (sex determining region y)-box9, collagen II and fibromodullin; and only to ADSC for collagen XI. BMSC was higher to ADSC for aggrecan and collagen IX, p ≤ 0.0001. The combination of TGF-β3 + BMP-6 revealed increased gene expressions on both BMSCs and ADSCs for early and mature chondrogenic markers, but no significance difference. For dedifferentiation markers, ADSC was significantly higher to chondrocyte for collagen I. Glycosaminoglycan evaluations with both formulas revealed that chondrocytes were significantly higher to ADSCs and BMSCs, but none was significant to each other, p ≤ 0.0001. Combination of 10 ng TGF-β3 with 10 ng of BMP-6 enhanced chondrogenic potentials of BMSCs and ADSCs compare to TGF-β3 alone. This could be the ideal cocktail for either cell’s chondrogenic induction.

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Acknowledgements

The authors wish to acknowledge the sponsorship by the Research University Grant/UKM-AP2011-26, DPP-2013-084 from the Ministry of Higher Education of Malaysia and DIP-2012-08.

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Authors and Affiliations

  1. Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia

    B. S. Aminuddin & B. H. I. Ruszymah

  2. Bioartificial Organ and Regenerative Medicine Unit, National Defence University of Malaysia, Sungai Besi, Camp, 57000, Kuala Lumpur, Malaysia

    C. C. Ude & B. M. Azizi

  3. Department of Clinical Veterinary, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Malaysia

    H. C. Chen

  4. Department of Orthopedic and Traumatology, Universiti Kebangsaan Malaysia Medical Center, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia

    M. Y. Norhamdan

  5. ENT Consultant Clinic, Ampang Putri Specialist Hospital, 68000, Ampang, Malaysia

    B. S. Aminuddin

  6. Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, 56000, Kuala Lumpur, Malaysia

    B. H. I. Ruszymah

Authors
  1. C. C. Ude
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  2. H. C. Chen
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  3. M. Y. Norhamdan
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  4. B. M. Azizi
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  5. B. S. Aminuddin
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  6. B. H. I. Ruszymah
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Correspondence to B. H. I. Ruszymah.

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The authors wish to declare that there was no conflict of interest during the study.

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Ude, C.C., Chen, H.C., Norhamdan, M.Y. et al. The evaluation of cartilage differentiations using transforming growth factor beta3 alone and with combination of bone morphogenetic protein-6 on adult stem cells. Cell Tissue Bank 18, 355–367 (2017). https://doi.org/10.1007/s10561-017-9638-1

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  • DOI: https://doi.org/10.1007/s10561-017-9638-1

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