A Novel Strategy for Preparation of ADSCs-Osteogenic Microtissue Based on CEFFE and Mechanisms for Enhancing Osteogenic Activity

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

The FASEB Journal Pub Date : 2025-09-01 DOI:10.1096/fj.202502341R

Mengqi Nie, Xizhe Li, Yutian Xiao, Shaorong Lei, Dingyu Wu

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Abstract

Adipose-derived mesenchymal stem cells (ADSCs) are more accessible than bone marrow mesenchymal stromal cells but have limited osteogenic differentiation capabilities in bone tissue engineering. Cell-free fat extract (CEFFE) from adipose tissue shows promise in enhancing the osteogenesis of ADSCs. This study aimed to investigate the impact of CEFFE on ADSCs and its role in promoting ADSC osteogenic activity both in vitro and in vivo. CEFFE was obtained and characterized. In vitro, ADSCs were isolated, characterized, and analyzed using various assays. Osteogenic differentiation was quantified through staining and gene expression analysis. ADSC-based osteogenic microtissues with CEFFE were created and tested in nude mice models. Molecular mechanisms were explored through transcriptomic analysis and western blotting. CEFFE enhanced various osteogenic aspects of ADSCs, including proliferation, calcium nodule formation, and expression of osteogenic markers. CEFFE-treated ADSCs showed early dense osteogenic cell sheet formation and increased ectopic bone volume in mice. Transcriptomic analysis revealed upregulation/downregulation of genes and activation of the PI3K-Akt signaling pathway by CEFFE. Western blotting confirmed that the PI3K-Akt pathway played a crucial role in CEFFE-induced acceleration of osteogenesis in ADSCs. The study confirms CEFFE's significant enhancement of ADSCs osteogenic potential and sheds light on the underlying mechanisms. These findings provide a preclinical foundation for potential applications in bone tissue engineering and regenerative medicine.

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基于CEFFE制备adscs -成骨微组织的新策略及增强成骨活性的机制

脂肪源性间充质干细胞（ADSCs）比骨髓间充质基质细胞更容易获得，但在骨组织工程中的成骨分化能力有限。从脂肪组织中提取的无细胞脂肪提取物（CEFFE）在促进ADSCs成骨方面表现出良好的前景。本研究旨在探讨CEFFE对ADSC的影响及其在体外和体内促进ADSC成骨活性的作用。制备了CEFFE并对其进行了表征。在体外，ADSCs被分离、表征，并使用各种方法进行分析。通过染色和基因表达分析量化成骨分化。建立了基于adsc的CEFFE成骨微组织，并在裸鼠模型中进行了测试。通过转录组学分析和western blotting研究其分子机制。CEFFE增强了ADSCs成骨的各个方面，包括增殖、钙结节的形成和成骨标志物的表达。ceffe处理的ADSCs在小鼠中显示早期致密成骨细胞片形成和异位骨体积增加。转录组学分析显示，CEFFE上调/下调基因，激活PI3K-Akt信号通路。Western blotting证实，PI3K-Akt通路在ceffe诱导的ADSCs加速成骨过程中发挥了关键作用。该研究证实了CEFFE对ADSCs成骨潜能的显著增强，并揭示了其潜在机制。这些发现为骨组织工程和再生医学的潜在应用提供了临床前基础。

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来源期刊

The FASEB Journal 生物-生化与分子生物学

CiteScore

9.20

自引率

2.10%

发文量

6243

审稿时长

3 months

期刊介绍： The FASEB Journal publishes international, transdisciplinary research covering all fields of biology at every level of organization: atomic, molecular, cell, tissue, organ, organismic and population. While the journal strives to include research that cuts across the biological sciences, it also considers submissions that lie within one field, but may have implications for other fields as well. The journal seeks to publish basic and translational research, but also welcomes reports of pre-clinical and early clinical research. In addition to research, review, and hypothesis submissions, The FASEB Journal also seeks perspectives, commentaries, book reviews, and similar content related to the life sciences in its Up Front section.