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
ARTICLE HIGHLIGHTS
Research background

Genetic manipulation is now considered the safest and promising approach in the field of regenerative medicine. Mesenchymal stem cells (MSCs) are the potential candidates for the clinical use of genetically engineered stem cells. They are non-immunogenic, proliferative, and possess multiple lineage differentiation potential. Gene modification of MSCs using chondrogenic transcription factors may lead to the use of this strategy to treat debilitating diseases related to cartilage.

Research motivation

Genetic modification of MSCs offers a competent source for the sustained translation of therapeutic proteins. Transfection of human umbilical cord derived MSCs (hUC-MSCs) using chondro-specific transcription factors may lead to enhance chondrogenesis and efficient regeneration of cartilage-related injuries.

Research objectives

The main objective of the study was to analyze the effect of transcription factors Sox-9 and Six-1 in chondrogenesis by transfecting them into hUC-MSCs and determining successful intervertebral disc (IVD) regeneration.

Research methods

MSCs were isolated from cord tissue and characterized using their specific markers. MSCs were transfected using transcription factors Sox-9 and Six-1. Cell differentiation was analyzed at the transcriptional and translational levels, while the regeneration potential of transfected MSCs was observed by transplanting them into the degenerated rat IVD model. Post-transplantation histology and cell tracking analysis were performed to evaluate cartilage regeneration.

Research results

In vitro analysis showed that transcription and translation of chondrogenic markers were significantly higher in transfected MSCs at 24 h, and 21 d in comparison to control MSCs. Transfected MSCs at the site of degenerated IVD differentiated into chondrocytes which secreted chondro-proteins. The cells homed and regenerated the injured cartilage as that of normal cartilage, as evident from immunohistological and histological analyses.

Research conclusions

Genetic modification of hUC-MSCs with two chondrogenic transcriptional factors Sox-9 and Six-1 enhances their chondrogenic differentiation. Their synergistic effect on MSCs accelerated the regeneration of degenerated cartilage with complete restoration of tissue architecture.

Research perspectives

The present manuscript offers a promising therapeutic approach to revolutionize the treatment of cartilaginous defects and spinal cord injuries. The outcomes discussed in this study accentuates the advances towards the clinical translation of such approaches.