Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jan 26, 2021; 13(1): 115-127
Published online Jan 26, 2021. doi: 10.4252/wjsc.v13.i1.115
Proliferation and tenogenic differentiation of bone marrow mesenchymal stem cells in a porous collagen sponge scaffold
Bing-Yu Zhang, Pu Xu, Qing Luo, Guan-Bin Song
Bing-Yu Zhang, Department of College of Bioinformatics, Chongqing University of Posts and Telecommunications, Chongqing 400065, China
Pu Xu, Qing Luo, Guan-Bin Song, Department of College of Bioengineering, Chongqing University, Chongqing 400030, China
Author contributions: Song GB, Zhang BY, and Luo Q conceived and designed the study; Zhang BY and Xu P performed the experiments; Zhang BY analyzed the data and wrote the manuscript; Song GB supervised the study; all authors reviewed and revised the manuscript critically.
Supported by Natural National Science Foundation of China, No. 31700810 and No. 11772073; Science and Technology Research Program of Chongqing Municipal Education Commission, No. KJQN201800601; Natural Science Foundation of Chongqing, China, No. cstc2020jcyj-msxmX0760; Visiting Scholar Foundation of Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, No. CQKLBST-2018-007.
Institutional animal care and use committee statement: The study was reviewed and approved by the Chongqing University of Posts and Telecommunications Institutional Review Board (No. CQUPT2018016).
Conflict-of-interest statement: The authors declare no potential conflicts of interest with respect to research, authorship, and/or publication of this article.
ARRIVE guidelines statement: The authors have read the ARRIVE Guidelines, and the manuscript was prepared and revised according to the ARRIVE Guidelines.
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:
Corresponding author: Guan-Bin Song, PhD, Professor, Department of College of Bioengineering, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400030, China.
Received: August 11, 2020
Peer-review started: August 11, 2020
First decision: October 23, 2020
Revised: November 2, 2020
Accepted: November 17, 2020
Article in press: November 17, 2020
Published online: January 26, 2021
Research background

How to manage the damaged tendon is still one of the most challenging problems in orthopedics. With the development of cell culture techniques, medical biomaterials, and implantation techniques, tissue engineering techniques show great potential for tendon repair and regeneration. Type I collagen is a popular natural materials used for tendon tissue engineering. One of the possible practical ways to further enhance tendon repair is to combine a porous collagen sponge scaffold with a suitable growth factor that has an inherent ability to promote the recruitment, proliferation, and tenogenic differentiation of cells.

Research motivation

At present, there is an incomplete understanding of which growth factors are sufficient and optimal for the tenogenic differentiation of stem cells in a porous collagen sponge scaffold.

Research objectives

To identify one or more ideal growth factors that benefit the proliferation or/and tenogenic differentiation of stem cells in a porous collagen sponge scaffold, which will help us better understand the role of growth factors in tendon tissue engineering.

Research methods

We constructed a 3D culture system based on a type I collagen sponge which is a porous scaffold material. The surface topography of the collagen sponge was observed by scanning electron microscopy. Cell survival on the surfaces of the collagen sponge was assessed by live/dead assay, and the activity of cells was assessed by CCK-8 assay. The mRNA and protein expression levels of related molecules were confirmed by quantitative real-time polymerase chain reaction and Western blot, respectively. The deposited collagen was assessed by Sirius Red staining. Histological analyses were performed to investigate the cell distribution and collagen deposition, which can help us visually understand the changes within the constructs.

Research results

Medical type I collagen sponge had a good biocompatibility. Transforming growth factor β1 (TGF-β1) had great promise for the tenogenic differentiation and proliferation of bone marrow mesenchymal stem cells (BMSCs) compared to growth differentiation factor 7 (GDF-7) and insulin-like growth factor 1 (IGF-1) in a monolayer culture (2D) and in a collagen sponge-based 3D culture. Moreover, TGF-β1 promoted more collagen deposition in both the 2D and 3D cultures. In the 2D culture, the proliferation of the BMSCs showed no significant changes compared to the control group after TGF-β1, IGF-1, or GDF-7 treatment. However, TGF-β1 and GDF-7 could increase the cell proliferation in the collagen sponge-based 3D culture. Strangely, we also found more dead cells in the 3D culture where cells were treated with TGF-β1.

Research conclusions

TGF-β1 shows great promise for BMSCs compared to GDF-7 and IGF-1 in both 2D and 3D cultures, and the collagen sponge-based 3D culture enhances the tenogenic differentiation of BMSCs well beyond the level of induction in the 2D culture after TGF-β1 treatment.

Research perspectives

Based on our results, we believe that TGF-β1 is an ideal growth factor that benefits the tenogenic differentiation of BMSCs in a porous collagen sponge scaffold.