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©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
Self-assembly of differentiated dental pulp stem cells facilitates spheroid human dental organoid formation and prevascularization
Fei Liu, Jing Xiao, Lei-Hui Chen, Yu-Yue Pan, Jun-Zhang Tian, Zhi-Ren Zhang, Xiao-Chun Bai
Fei Liu, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong Province, China
Fei Liu, Jun-Zhang Tian, Department of Health Management, Guangdong Second Provincial General Hospital, Guangzhou 510317, Guangdong Province, China
Jing Xiao, Guangdong Provincial Key Laboratory of Tumor Interventional Diagnosis and Treatment, Zhuhai People’s Hospital Affiliated with Jinan University, Zhuhai 519000, Guangdong Province, China
Jing Xiao, Centre of Reproduction, Development and Aging, Faculty of Health Sciences, University of Macau, Macau 999078, China
Lei-Hui Chen, Yu-Yue Pan, Department of Stomatology, Guangdong Second Provincial General Hospital, Guangzhou 510317, Guangdong Province, China
Zhi-Ren Zhang, Zhuhai Institute of Translational Medicine, Zhuhai Hospital Affiliated with Jinan University, Zhuhai 519000, Guangdong Province, China
Xiao-Chun Bai, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong Province, China
Co-first authors: Fei Liu and Jing Xiao.
Co-corresponding authors: Zhi-Ren Zhang and Chun-Xiao Bai.
Author contributions: Liu F, Xiao J, Tian JZ, Zhang ZR, and Bai XC designed the research; Liu F and Xiao J performed the research and analyzed the data; Chen LH and Pan YY contributed human dental pulp tissue; Liu F wrote the paper. Liu F and Xiao J contributed to the work equally and should be regarded as co-first authors. Bai XC and Zhang ZR contributed to the work equally and should be regarded as co-corresponding author.
Supported by the Science and Technology Programme of Guangzhou City, No. 202201020341.
Institutional review board statement: This study was approved by the Ethics Committee of Guangdong Second Provincial General Hospital.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at kqliufei@126.com. Participants gave informed consent for data sharing.
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: Xiao-Chun Bai, PhD, Professor, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, No. 1023-1063 Shatai South Road, Baiyun District, Guangzhou 510515, Guangdong Province, China. baixc15@smu.edu.cn
Received: December 19, 2023
Peer-review started: December 19, 2023
First decision: January 12, 2024
Revised: January 21, 2024
Accepted: February 28, 2024
Article in press: February 28, 2024
Published online: March 26, 2024
Processing time: 96 Days and 18.9 Hours
BACKGROUND
The self-assembly of solid organs from stem cells has the potential to greatly expand the applicability of regenerative medicine. Stem cells can self-organise into microsized organ units, partially modelling tissue function and regeneration. Dental pulp organoids have been used to recapitulate the processes of tooth development and related diseases. However, the lack of vasculature limits the utility of dental pulp organoids.
AIM
To improve survival and aid in recovery after stem cell transplantation, we demonstrated the three-dimensional (3D) self-assembly of adult stem cell-human dental pulp stem cells (hDPSCs) and endothelial cells (ECs) into a novel type of spheroid-shaped dental pulp organoid in vitro under hypoxia and conditioned medium (CM).
METHODS
During culture, primary hDPSCs were induced to differentiate into ECs by exposing them to a hypoxic environment and CM. The hypoxic pretreated hDPSCs were then mixed with ECs at specific ratios and conditioned in a 3D environment to produce prevascularized dental pulp organoids. The biological characteristics of the organoids were analysed, and the regulatory pathways associated with angiogenesis were studied.
RESULTS
The combination of these two agents resulted in prevascularized human dental pulp organoids (Vorganoids) that more closely resembled dental pulp tissue in terms of morphology and function. Single-cell RNA sequencing of dental pulp tissue and RNA sequencing of Vorganoids were integrated to analyse key regulatory pathways associated with angiogenesis. The biomarkers forkhead box protein O1 and fibroblast growth factor 2 were identified to be involved in the regulation of Vorganoids.
CONCLUSION
In this innovative study, we effectively established an in vitro model of Vorganoids and used it to elucidate new mechanisms of angiogenesis during regeneration, facilitating the development of clinical treatment strategies.
Core Tip: We demonstrated the three-dimensional self-assembly of adult stem cell-human dermal papilla cells and endothelial cells into a novel type of spheroid-shaped dental pulp organoid in vitro under hypoxia and conditioned medium. These organoids have been constructed to be morphologically and functionally closer to dental pulp tissue. Through the integration and analysis of single-cell RNA sequencing and RNA sequencing data, forkhead box protein O1 and fibroblast growth factor 2 were identified as crucial markers involved in the regulation of organoid angiogenesis. In this innovative study, we effectively established an in vitro model of prevascularized dental pulp organoids and used it to elucidate new mechanisms of angiogenesis during regeneration, facilitating the development of clinical treatment strategies.
