Copyright
©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
Calcifying nanoparticles induce apoptosis and calcification in bone marrow mesenchymal stem cells via the transforming growth factor-β/Smad pathway
Xuan-Li Su, Fu-Rong Xu, Jian Yang, San-Qiang Niu, Hao-Jie Shi, Yu-Fan He, Zhen-Hao Li, Pankaj Bagari, Xiang-Wei Wu, Xin-Yu Peng, Hong-Wei Zhang, Mei-Yan Wang
Xuan-Li Su, Department of Surgery, The First Affiliated Hospital of Shihezi University, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Fu-Rong Xu, Jian Yang, San-Qiang Niu, Department of Emergency, The First Affiliated Hospital of Shihezi University, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Hao-Jie Shi, Pankaj Bagari, Hong-Wei Zhang, Department of General Surgery, The First Affiliated Hospital of Shihezi University, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Yu-Fan He, Department of General Practice, The First Affiliated Hospital of Shihezi University, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Zhen-Hao Li, The First Affiliated Hospital of Shihezi University, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Xiang-Wei Wu, Department of Administrative, Shihezi University School of Medicine, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Xin-Yu Peng, Department of Administrative, The First Affiliated Hospital of Shihezi University, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Mei-Yan Wang, Medicine Academy, Shihezi University School of Medicine, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
Co-first authors: Xuan-Li Su and Fu-Rong Xu.
Author contributions: Su XL and Xu FR contributed equally to this manuscript and are co-first authors. Su XL mainly contributed to drafting the article, critical revisions, and data interpretation; Xu FR and Yang J equally edited and reviewed the manuscript; Yang J contributed with final approval for publication; Niu SQ, Shi HJ, He YF, Li ZH, Bagari P, Wu XW, Peng XY, Zhang HW, and Wang MY approved the final article.
Supported by the Project of Xinjiang Production and Construction Corps, No. 2022ZD090; the Project of Xinjiang Production and Construction Corps - Young Science and Technology Innovation Talents, No. 2023CB008-31; The First Affiliated Hospital of Shihezi University Medical College, Doctoral Fund Project, No. BS202207; Talent Development Fund-Tianshan Talents, No. CZ001219; and 2024 National Health Commission Central Asian High-Incidence Prevention and Control Key Laboratory, No. KF202405.
Institutional review board statement: This study used commercially available cell lines and did not involve human or animal subjects, thus IRB approval was not required.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The materials and data used in this study are available from the corresponding author upon reasonable request.
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: Jian Yang, Professor, Department of Emergency, The First Affiliated Hospital of Shihezi University, North Fourth Road, Shihezi 832000, Xinjiang Uygur Autonomous Region, China.
125210525@qq.com
Received: March 3, 2025
Revised: May 22, 2025
Accepted: November 10, 2025
Published online: December 26, 2025
Processing time: 299 Days and 9.9 Hours
BACKGROUND
Pathological calcification is a common feature of many diseases. Calcifying nanoparticles (CNPs) are considered potential inducers of this abnormal calcification, but their specific effects on bone marrow mesenchymal stem cells (BMSCs) remain unclear. BMSCs are key cells in bone formation and repair, and their aberrant apoptosis and calcification are closely related to disease progression.
AIM
To explore whether CNPs can induce apoptosis and calcification in BMSCs and analyzed the relationship between these processes. The differential effects of CNPs and nanoscale hydroxyapatites (nHAPs) in inducing apoptosis and calcification in BMSCs were also compared.
METHODS
CNPs obtained in the early stage were identified by electron microscopy and particle size analysis. BMSCs were cultured with various treatments, including different concentrations of nHAPs, CNPs [2 McFarland (MCF) turbidity, 4 MCF, 6 MCF], and a transforming growth factor (TGF)-β inhibitor (SB431542) for 72 hours. The isolated CNPs exhibited the expected sizes and shapes.
RESULTS
Exposure to CNPs and nHAPs suppressed cell proliferation and promoted apoptosis in a concentration-dependent manner, with CNPs exhibiting significantly stronger effects. Alizarin Red staining indicated an increase in calcium deposition with exposure to increasing concentrations of nHAPs and CNPs. Quantitative reverse-transcription polymerase chain reaction results indicated that medium concentrations of nHAPs and CNPs significantly enhanced the expression of pro-apoptotic and pro-calcification markers, whereas the expression of anti-apoptotic Bcl-2 was reduced compared with untreated controls. Western blotting results showed that medium concentrations of CNPs and nHAPs increased the expression of osteopontin, bone morphogenetic protein-2, TGF-β/Smad, Bax, and caspase-3 and decreased Bcl-2 expression compared with controls.
CONCLUSION
CNPs and nHAPs induced apoptosis and calcification in BMSCs, with CNPs being the most potent. Additionally, the TGF-β inhibitor SB431542 significantly reduced the occurrence of apoptosis and calcification. A correlation was found between apoptosis and calcification, which is likely mediated through the TGF-β/Smad signaling pathway.
Core Tip: This study revealed calcifying nanoparticles (CNPs) induce cell apoptosis and calcification on bone marrow mesenchymal stem cells. Through in vitro experiments, we found that CNPs inhibit cell proliferation in a concentration dependent manner and significantly enhance the expression of pro-apoptotic and pro-calcification markers. In addition, the transforming growth factor-β/Smad signaling pathway plays a crucial role in this process. These findings provide a new perspective for understanding the potential applications of CNPs in tissue repair and regenerative medicine.
