Enhancing Bone Formation Through bFGF-Loaded Mesenchymal Stromal Cell Spheroids During Fracture Healing in Mice.

IF 3.8 3区 医学 Q2 ENGINEERING, BIOMEDICAL
Kugo Takeda, Hiroki Saito, Shintaro Shoji, Hiroyuki Sekiguchi, Mitsuyoshi Matsumoto, Masanobu Ujihira, Masayuki Miyagi, Gen Inoue, Masashi Takaso, Kentaro Uchida
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引用次数: 0

Abstract

This study aimed to evaluate the osteogenic potential of mesenchymal stromal cell (MSC) spheroids combined with the basic fibroblast growth factor (bFGF) in a mouse femur fracture model. To begin, MSC spheroids were generated, and the expression of key trophic factors (bFGF Bmp2, and Vegfa) was assessed using quantitative PCR (qPCR). A binding assay confirmed the interaction between the bFGF and the spheroids' extracellular matrix. The spheroid cultures significantly upregulated bFGF, Bmp2, and Vegfa expression compared to the monolayers (p < 0.001), and the binding assay demonstrated effective bFGF binding to the MSC spheroids. Following these in vitro assessments, the mice were divided into five groups for the in vivo study: (1) no treatment (control), (2) spheroids alone, (3) bFGF alone, (4) bFGF-loaded spheroids (bFGF-spheroids), and (5) non-viable (frozen) bFGF-loaded spheroids (bFGF-dSpheroids). Bone formation was analyzed by a micro-CT, measuring the bone volume (BV) and bone mineral content (BMC) of the mice four weeks post-fracture. A high dose of the bFGF (10 µg) significantly promoted bone formation regardless of the presence of spheroids, as evidenced by the increases in BV (bFGF, p = 0.010; bFGF-spheroids, p = 0.006; bFGF-dSpheroids, p = 0.032) and BMC (bFGF, p = 0.023; bFGF-spheroids, p = 0.004; bFGF-dSpheroids, p = 0.014), compared to the controls. In contrast, a low dose of the bFGF (1 µg) combined with the MSC spheroids significantly increased BV and BMC compared to the control (BV, p = 0.012; BMC, p = 0.015), bFGF alone (BV, p = 0.012; BMC, p = 0.008), and spheroid (BV, p < 0.001; BMC, p < 0.001) groups. A low dose of the bFGF alone did not significantly promote bone formation (p > 0.05). The non-viable (frozen) spheroids loaded with a low dose of the bFGF resulted in a higher BV and BMC compared to the spheroids alone (BV, p = 0.003; BMC, p = 0.017), though the effect was less pronounced than in the viable spheroids. These findings demonstrate the synergistic effect of the bFGF and MSC spheroids on bone regeneration. The increased expression of the BMP-2 and VEGF observed in the initial experiments, coupled with the enhanced bone formation in vivo, highlight the therapeutic potential of this combination. Future studies will aim to elucidate the underlying molecular mechanisms and assess the long-term outcomes for bone repair strategies.

在小鼠骨折愈合过程中通过 bFGF/Loaded 间充质基质细胞球体促进骨形成
本研究旨在评估间充质基质细胞(MSC)球形体结合碱性成纤维细胞生长因子(bFGF)在小鼠股骨骨折模型中的成骨潜力。首先,生成间充质干细胞球体,并使用定量 PCR(qPCR)评估关键营养因子(bFGF、Bmp2 和 Vegfa)的表达。结合试验证实了 bFGF 与球体细胞外基质之间的相互作用。与单层培养物相比,球形培养物的 bFGF、Bmp2 和 Vegfa 表达明显升高(p < 0.001),结合试验也证明了 bFGF 与间充质干细胞球体的有效结合。体外评估结束后,小鼠被分为五组进行体内研究:(1) 无处理(对照组),(2) 单用球形细胞,(3) 单用 bFGF,(4) bFGF 负载球形细胞(bFGF-spheroids),(5) 无活力(冷冻)bFGF 负载球形细胞(bFGF-dSpheroids)。通过显微 CT 分析骨形成情况,测量小鼠骨折后四周的骨量(BV)和骨矿物质含量(BMC)。高剂量的 bFGF(10 µg)能显著促进骨形成,无论是否存在球形体,BV 的增加就是证明(bFGF,p = 0.010; bFGF-spheroids, p = 0.006; bFGF-dSpheroids, p = 0.032)和 BMC(bFGF, p = 0.023; bFGF-spheroids, p = 0.004; bFGF-dSpheroids, p = 0.014)的增加。相反,与对照组(BV,p = 0.012;BMC,p = 0.015)、单用 bFGF 组(BV,p = 0.012;BMC,p = 0.008)和球形组(BV,p < 0.001;BMC,p < 0.001)相比,低剂量 bFGF(1 µg)与间充质干细胞球体结合可显著增加 BV 和 BMC。单用低剂量的 bFGF 并不能显著促进骨形成(p > 0.05)。与单独的球形体相比,加载了低剂量 bFGF 的非存活(冷冻)球形体具有更高的 BV 和 BMC(BV,p = 0.003;BMC,p = 0.017),但效果不如存活球形体明显。这些发现证明了 bFGF 和间充质干细胞球体对骨再生的协同作用。初步实验中观察到的 BMP-2 和血管内皮生长因子的表达增加,加上体内骨形成的增强,凸显了这种组合的治疗潜力。未来的研究将旨在阐明潜在的分子机制,并评估骨修复策略的长期效果。
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来源期刊
Bioengineering
Bioengineering Chemical Engineering-Bioengineering
CiteScore
4.00
自引率
8.70%
发文量
661
期刊介绍: Aims Bioengineering (ISSN 2306-5354) provides an advanced forum for the science and technology of bioengineering. It publishes original research papers, comprehensive reviews, communications and case reports. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. All aspects of bioengineering are welcomed from theoretical concepts to education and applications. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. There are, in addition, four key features of this Journal: ● We are introducing a new concept in scientific and technical publications “The Translational Case Report in Bioengineering”. It is a descriptive explanatory analysis of a transformative or translational event. Understanding that the goal of bioengineering scholarship is to advance towards a transformative or clinical solution to an identified transformative/clinical need, the translational case report is used to explore causation in order to find underlying principles that may guide other similar transformative/translational undertakings. ● Manuscripts regarding research proposals and research ideas will be particularly welcomed. ● Electronic files and software regarding the full details of the calculation and experimental procedure, if unable to be published in a normal way, can be deposited as supplementary material. ● We also accept manuscripts communicating to a broader audience with regard to research projects financed with public funds. Scope ● Bionics and biological cybernetics: implantology; bio–abio interfaces ● Bioelectronics: wearable electronics; implantable electronics; “more than Moore” electronics; bioelectronics devices ● Bioprocess and biosystems engineering and applications: bioprocess design; biocatalysis; bioseparation and bioreactors; bioinformatics; bioenergy; etc. ● Biomolecular, cellular and tissue engineering and applications: tissue engineering; chromosome engineering; embryo engineering; cellular, molecular and synthetic biology; metabolic engineering; bio-nanotechnology; micro/nano technologies; genetic engineering; transgenic technology ● Biomedical engineering and applications: biomechatronics; biomedical electronics; biomechanics; biomaterials; biomimetics; biomedical diagnostics; biomedical therapy; biomedical devices; sensors and circuits; biomedical imaging and medical information systems; implants and regenerative medicine; neurotechnology; clinical engineering; rehabilitation engineering ● Biochemical engineering and applications: metabolic pathway engineering; modeling and simulation ● Translational bioengineering
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