Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Feb 26, 2022; 14(2): 163-182
Published online Feb 26, 2022. doi: 10.4252/wjsc.v14.i2.163
Transcription regulators differentiate mesenchymal stem cells into chondroprogenitors, and their in vivo implantation regenerated the intervertebral disc degeneration
Shumaila Khalid, Sobia Ekram, Asmat Salim, G. Rasul Chaudhry, Irfan Khan
Shumaila Khalid, Sobia Ekram, Asmat Salim, Irfan Khan, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Sindh, Pakistan
G. Rasul Chaudhry, Department of Biological Sciences, Oakland University, Rochester, MI 48309, United States
Author contributions: Khalid S performed the experiment and wrote the first draft of manuscript; Ekram S helped in experimentation and data acquisition; Salim A and Chaudhry GR evaluated the data and helped in manuscript preparation; Khan I designed the experiment, evaluated and analyzed the data, secured the funding, and finalized the manuscript.
Supported by Higher Education Commission Pakistan, No. 7083.
Institutional review board statement: IEC approval for the protocol ICCBS/IEC-009-UCB-2015/protocol/1.0. Informed consent was obtained from donor parents for the use of umbilical cord tissue in research.
Institutional animal care and use committee statement: Approval for Animal Study Protocol, No. 2018-0016.
Conflict-of-interest statement: No conflict of interest.
Data sharing statement: No extra data to share.
ARRIVE guidelines statement: ARRIVE guidelines were followed.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Irfan Khan, PhD, Assistant Professor, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, KU Circular Rd, Karachi 75270, Sindh, Pakistan. khan@iccs.edu
Received: February 28, 2021
Peer-review started: February 28, 2021
First decision: April 19, 2021
Revised: May 2, 2021
Accepted: January 6, 2022
Article in press: January 6, 2022
Published online: February 26, 2022
Processing time: 362 Days and 2.9 Hours
Abstract
BACKGROUND

Intervertebral disc degeneration (IVDD) is the leading cause of lower back pain. Disc degeneration is characterized by reduced cellularity and decreased production of extracellular matrix (ECM). Mesenchymal stem cells (MSCs) have been envisioned as a promising treatment for degenerative illnesses. Cell-based therapy using ECM-producing chondrogenic derivatives of MSCs has the potential to restore the functionality of the intervertebral disc (IVD).

AIM

To investigate the potential of chondrogenic transcription factors to promote differentiation of human umbilical cord MSCs into chondrocytes, and to assess their therapeutic potential in IVD regeneration.

METHODS

MSCs were isolated and characterized morphologically and immunologically by the expression of specific markers. MSCs were then transfected with Sox-9 and Six-1 transcription factors to direct differentiation and were assessed for chondrogenic lineage based on the expression of specific markers. These differentiated MSCs were implanted in the rat model of IVDD. The regenerative potential of transplanted cells was investigated using histochemical and molecular analyses of IVDs.

RESULTS

Isolated cells showed fibroblast-like morphology and expressed CD105, CD90, CD73, CD29, and Vimentin but not CD45 antigens. Overexpression of Sox-9 and Six-1 greatly enhanced the gene expression of transforming growth factor beta-1 gene, BMP, Sox-9, Six-1, and Aggrecan, and protein expression of Sox-9 and Six-1. The implanted cells integrated, survived, and homed in the degenerated intervertebral disc. Histological grading showed that the transfected MSCs regenerated the IVD and restored normal architecture.

CONCLUSION

Genetically modified MSCs accelerate cartilage regeneration, providing a unique opportunity and impetus for stem cell-based therapeutic approach for degenerative disc diseases.

Keywords: Intervertebral disc degeneration; Human umbilical cord; Transcription factors; Mesenchymal stem cells; Gene expression; Regeneration

Core Tip: In this study, we highlighted that overexpression of chondrogenic transcription factors in human umbilical cord derived mesenchymal stem cells (hUC-MSCs) accelerated their differentiation into chondroprogenitor cells. The synergistic effect of Sox-9 and Six-1 transcription factors leads the MSCs to differentiate into chondrogenic cells in the basal medium, which produced the same effect as the chondro-induction medium. In vivo transplantation of these transfected cells leads to their homing, integration, and differentiation into nucleus pulposus cells of the intervertebral disc. This approach could help to develop a better treatment option for degenerative disc diseases.