骨形态发生蛋白9（BMP9）和神经生长因子（NGF）对C3H10T1/2间充质干细胞成骨分化的协同影响

Synergistic influences of BMP9 and NGF on the osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.

作者信息

Liu Junyu, Yang Kun, Li Gang, Tan Yinghui

机构信息

Chongqing Key Laboratory of Oral Diseases, The Affiliated Stomatological Hospital of Chongqing Medical University, No.426 Songshi North Road, Yubei District, Chongqing, 401147, China.

Department of Periodontology, Affiliated Stomatology Hospital of Zunyi Medical University, No.149 Dalian Road, Huichuan District, Zunyi City, Guizhou Province, 563099, China.

出版信息

J Orthop Surg Res. 2025 Mar 15;20(1):287. doi: 10.1186/s13018-025-05669-4.

DOI:10.1186/s13018-025-05669-4

PMID:40089726

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11909987/

Abstract

BACKGROUND

Bone morphogenetic protein 9 (BMP9) and nerve growth factor (NGF) are critical factors influencing osteogenic differentiation in mesenchymal stem cells (MSCs). While BMP9 has been recognized for its potent osteogenic capabilities, NGF's role in bone tissue engineering is less understood. This investigation delineated the synergistic link of BMP9 with NGF in driving osteogenic differentiation in C3H10T1/2 MSCs.

OBJECTIVE

To evaluate the combined impact of BMP9 and NGF on osteogenic markers' expression levels and the formation of calcified nodules in C3H10T1/2 cells, providing a basis for the enhanced bone regeneration strategies in tissue engineering.

METHODS

C3H10T1/2 cells were subjected to treatment regimens incorporating NGF at variable concentrations (10, 50, and 100 ng/ml) and BMP9, either as monotherapies or in combination. Osteogenic differentiation' comprehensive assessment was undertaken by quantifying early-stage markers (Runx2, Col I) and late-stage markers (OPN) via RT-PCR, Western blotting, ALP staining, and Alizarin Red S staining for mineralized matrix deposition.

RESULTS

NGF elicited a concentration-dependent augmentation of early osteogenic markers, with the 10 ng/ml dosage demonstrating maximal efficacy. BMP9 independently facilitated robust osteogenic differentiation, whereas the combinatorial treatment with BMP9 and NGF synergistically amplified the expression levels of Runx2, Col I, and OPN. Notably, this combined treatment yielded a remarkable enhancement in the deposition of mineralized extracellular matrix, as evidenced by a notable escalation in the size and density of calcified nodules relative to monotherapies.

CONCLUSION

The findings unveiled the remarkable synergistic link of BMP9 with NGF in potentiating osteogenic differentiation in C3H10T1/2 MSCs. This dual-factor approach presents a compelling paradigm for advancing bone regeneration strategies, providing substantial promise for utilization in bone tissue engineering plus regenerative medicine.

摘要

背景

骨形态发生蛋白9（BMP9）和神经生长因子（NGF）是影响间充质干细胞（MSCs）成骨分化的关键因素。虽然BMP9因其强大的成骨能力而被认可，但NGF在骨组织工程中的作用尚鲜为人知。本研究阐述了BMP9与NGF在驱动C3H10T1/2 MSCs成骨分化中的协同联系。

目的

评估BMP9和NGF对C3H10T1/2细胞中成骨标志物表达水平和钙化结节形成的联合影响，为组织工程中增强骨再生策略提供依据。

方法

C3H10T1/2细胞接受不同浓度（10、50和100 ng/ml）NGF与BMP9的单一疗法或联合疗法处理。通过逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法、碱性磷酸酶（ALP）染色以及茜素红S染色定量早期标志物（Runx2、I型胶原）和晚期标志物（骨桥蛋白，OPN），对成骨分化进行综合评估，以检测矿化基质沉积情况。

结果

NGF引起早期成骨标志物呈浓度依赖性增加，10 ng/ml剂量显示出最大效果。BMP9单独促进了强劲的成骨分化，而BMP9与NGF的联合处理协同放大了Runx2、I型胶原和OPN的表达水平。值得注意的是，与单一疗法相比，这种联合处理使矿化细胞外基质的沉积显著增加，表现为钙化结节的大小和密度明显升高。