Evaluation of the chondrogenic maturation of in vitro chondrogenically induced BMSCs and ADSCs after implantation in nude mice
-
1
Universiti Kebangsaan Malaysia Medical Centre, Tissue Engineering Centre, Malaysia
-
2
National Defence University of Malaysia, Bioartificial Organ Research and Regenerative Medicine Unit, Malaysia
-
3
Ampang Putri Specialist Hospital , ENT Consultant Clinic, Malaysia
-
4
Universiti Kebangsaan Malaysia Medical Centre, Department of Physiology, Faculty of Medicine , Malaysia
Bioengineering of mature functional tissues or organs has been the goal of tissue engineering and regenerative medicine. Over the years, researchers have devised the means of isolating stem cells from different tissues in the body as tools for the generation of similar or different tissues within a two dimensional (2D) culture flask in the laboratory1. This in vitro differentiations of isolated cells to tissues, lack the necessary clues obtainable in living body for the induced cells to become as mature as the native tissue. It has been postulated that the body aids implanted immature bioengineered tissues towards maturation in vivo 2.Hence, we evaluated the ability of the body to confer maturity to in vitro chondrogenically induced implanted bone marrow stem cells (BMSCs) and adipose stem cells (ADSCs) owing to the availability of these physiological clues. In this study, human BMSCs and ADSCs obtained from discarded redundant tissues were grown and differentiated to chondrogenic lineage in culture flasks. Thereafter, they were trypsinized, homogenised and seeded in 2ml fibrin (Fig 1). Constructs were made by the addition of calcium chloride in the fibrin/cell mixture, then they were implanted in the dorsum of nude mice, divided into 3 groups each as follows: uninduced BMSCs/ ADSCs; induced BMSCs/ ADSCs and native chondrocytes. Constructs were harvested at 6weeks post implantations for analysis. Pre-induction assessment showed that both cells were positive to cluster of differentiations CDs (44, 73, 90 and 105) and negative to CDs (10, 34, 45 & HLA-DR). Both in vitro induced ADSCs and BMSCs showed characteristic features of chondrogenesis. They were positive to toluidine blue and safranin O. Their immunohistochemistry showed positive to collagen I (col I), col II, agreccan and (sex determining region y)-box 9 (sox9) antigens. The in vitro chondrogenic gene analysis for cartilage oligomeric protein (COMP), sry (sex determining region y)-box 5 (sox5), pro-alpha col II, chondroadherin, agreccan, col ix, sox9, col II alpha 1, col xi, fibromodullin, col I and col x revealed that BMSCs had mRNA fold increases of (150, 15, 10, 10, 11, 2, 4, 0.2, 0.8, 31, 3500 and 2), while ADSCs had (300, 35, 80, 30, 300, 2, 6, 0.6, 0.5, 150, 1000 and 60) respectively. Post implantation gene analysis revealed that an up regulation of cartilage oligomeric protein, sox5, pro-alpha col II, chondroadherin, agreccan, col ix, sox9, col II alpha 1, col xi, fibromodullin, col I and col x for BMSCs were (450, 100, 80, 350, 150, 900, 250, 250, 80, 100, 1000 and 100), while ADSCs had (700, 100, 200, 400, 120, 1000, 300, 200, 40, 120, 500 and 80) respectively. Post implantation Safranin O and toluidine blue evaluation showed more maturity compare to the in vitro. Ultra-structure analysis from the scanning electron microscopy (SEM) of the groups showed that both induced ADSCs and BMSCs had close resemblance to the native tissue (Fig 2). The above result showed that the body retains the ability of a natural bioreactor. Hence, in vivo implantation could provide the necessary clues for the maturity of newly 2D engineered tissue. In conclusion, the implanted chondrogenic induced BMSCs and ADSCs showed significantly higher cartilage qualities compare to the in vitro analysis.
Acknowledgements
Sponsored by the Research University grant/ UKM-AP2011-26, DPP-2013-084 from the Ministry of Higher Education of Malaysia.
Keywords:
Cartilage,
Chondrogenesis,
Tissue Engineering,
stem cell,
Cells
Conference:
6th Malaysian Tissue Engineering and Regenerative Medicine Scientific Meeting (6th MTERMS) 2016 and 2nd Malaysian Stem Cell Meeting, Seberang Jaya, Penang, Malaysia, 17 Nov - 18 Nov, 2016.
Presentation Type:
Poster
Topic:
Stem cells and Regenerative Medicine
Citation:
Ude
CC,
Seet
WT,
S
SA,
Ng
MH,
Saim
AB and
Idrus
RH
(2016). Evaluation of the chondrogenic maturation of in vitro chondrogenically induced BMSCs and ADSCs after implantation in nude mice.
Front. Bioeng. Biotechnol.
Conference Abstract:
6th Malaysian Tissue Engineering and Regenerative Medicine Scientific Meeting (6th MTERMS) 2016 and 2nd Malaysian Stem Cell Meeting.
doi: 10.3389/conf.FBIOE.2016.02.00002
Copyright:
The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers.
They are made available through the Frontiers publishing platform as a service to conference organizers and presenters.
The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated.
Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed.
For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions.
Received:
08 Dec 2016;
Published Online:
19 Dec 2016.
*
Correspondence:
Mr. Chinedu C Ude, Universiti Kebangsaan Malaysia Medical Centre, Tissue Engineering Centre, Cheras, Selangor, 56000, Malaysia, nedutus@yahoo.com