Basic Study
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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: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Guan-Bin Song, PhD, Professor, Department of College of Bioengineering, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400030, China. song@cqu.edu.cn
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
Processing time: 162 Days and 15.3 Hours
Abstract
BACKGROUND

Collagen is one of the most commonly used natural biomaterials 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 or cytokine that has an inherent ability to promote the recruitment, proliferation, and tenogenic differentiation of cells. However, there is an incomplete understanding of which growth factors are sufficient and optimal for the tenogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in a collagen sponge-based 3D culture system.

AIM

To identify one or more ideal growth factors that benefit the proliferation and tenogenic differentiation of rat BMSCs in a porous collagen sponge scaffold.

METHODS

We constructed a 3D culture system based on a type I collagen sponge scaffold. The surface topography of the collagen sponge scaffold was observed by scanning electron microscopy. Primary BMSCs were isolated from Sprague-Dawley rats. Cell survival on the surfaces of the scaffolds with different growth factors was assessed by live/dead assay and CCK-8 assay. The mRNA and protein expression levels were confirmed by quantitative real-time polymerase chain reaction and Western blot, respectively. The deposited collagen was assessed by Sirius Red staining.

RESULTS

Transforming growth factor β1 (TGF-β1) showed great promise in the tenogenic differentiation of BMSCs compared to growth differentiation factor 7 (GDF-7) and insulin-like growth factor 1 (IGF-1) in both the 2D and 3D cultures, and the 3D culture enhanced the differentiation of BMSCs into tenocytes well beyond the level of induction in the 2D culture after TGF-β1 treatment. 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 3D culture. Strangely, we also found more dead cells in the BMSC-collagen sponge constructs that were treated with TGF-β1. Moreover, TGF-β1 promoted more collagen deposition in both the 2D and 3D cultures.

CONCLUSION

Collagen sponge-based 3D culture with TGF-β1 enhances the responsiveness of the proliferation and tenogenic differentiation of rat BMSCs.

Keywords: Bone marrow mesenchymal stem cells; Collagen sponge; Transforming growth factor β1; Tenogenic differentiation; Proliferation; Collagen deposition

Core Tip: Growth factor supplementation of stem cells facilitates cell differentiation and/or cell growth, but there is an incomplete understanding of which growth factors are optimal in 3D culture. We compared different growth factors for the proliferation and tenogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in a monolayer culture (2D) and in a collagen sponge-based 3D culture. We found that transforming growth factor β1 (TGF-β1) showed great promise in the tenogenic differentiation of BMSCs compared to growth differentiation factor 7 and insulin-like growth factor 1 in both 2D and 3D cultures, and the 3D culture enhanced the tenogenic differentiation of BMSCs well beyond the level of induction in the 2D culture after TGF-β1 treatment.