Enhancement of bone regeneration by coadministration of angiogenic and osteogenic factors using messenger RNA.

IF 5 3区 医学 Q2 IMMUNOLOGY Inflammation and Regeneration Pub Date : 2023-06-20 DOI:10.1186/s41232-023-00285-3
Maorui Zhang, Yuta Fukushima, Kosuke Nozaki, Hideyuki Nakanishi, Jia Deng, Noriyuki Wakabayashi, Keiji Itaka
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引用次数: 1

Abstract

Background: Bone defects remain a challenge today. In addition to osteogenic activation, the crucial role of angiogenesis has also gained attention. In particular, vascular endothelial growth factor (VEGF) is likely to play a significant role in bone regeneration, not only to restore blood supply but also to be directly involved in the osteogenic differentiation of mesenchymal stem cells. In this study, to produce additive angiogenic-osteogenic effects in the process of bone regeneration, VEGF and Runt-related transcription factor 2 (Runx2), an essential transcription factor for osteogenic differentiation, were coadministered with messenger RNAs (mRNAs) to bone defects in the rat mandible.

Methods: The mRNAs encoding VEGF or Runx2 were prepared via in vitro transcription (IVT). Osteogenic differentiation after mRNA transfection was evaluated using primary osteoblast-like cells, followed by an evaluation of the gene expression levels of osteogenic markers. The mRNAs were then administered to a bone defect prepared in the rat mandible using our original cationic polymer-based carrier, the polyplex nanomicelle. The bone regeneration was evaluated by micro-computerized tomography (μCT) imaging, and histologic analyses.

Results: Osteogenic markers such as osteocalcin (Ocn) and osteopontin (Opn) were significantly upregulated after mRNA transfection. VEGF mRNA was revealed to have a distinct osteoblastic function similar to that of Runx2 mRNA, and the combined use of the two mRNAs resulted in further upregulation of the markers. After in vivo administration into the bone defect, the two mRNAs induced significant enhancement of bone regeneration with increased bone mineralization. Histological analyses using antibodies against the Cluster of Differentiation 31 protein (CD31), alkaline phosphatase (ALP), or OCN revealed that the mRNAs induced the upregulation of osteogenic markers in the defect, together with increased vessel formation, leading to rapid bone formation.

Conclusions: These results demonstrate the feasibility of using mRNA medicines to introduce various therapeutic factors, including transcription factors, into target sites. This study provides valuable information for the development of mRNA therapeutics for tissue engineering.

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利用信使RNA联合使用血管生成因子和成骨因子促进骨再生。
背景:骨缺损在今天仍然是一个挑战。除了成骨激活外,血管生成的关键作用也引起了人们的关注。特别是血管内皮生长因子(VEGF)可能在骨再生中发挥重要作用,不仅恢复血液供应,而且直接参与间充质干细胞的成骨分化。在本研究中,为了在骨再生过程中产生加性的血管生成-成骨作用,我们将VEGF和成骨分化必需的转录因子runt相关转录因子2 (Runx2)与信使rna (mrna)共同给予大鼠下颌骨骨缺损。方法:采用体外转录(IVT)法制备编码VEGF或Runx2的mrna。利用原代成骨细胞样细胞评估mRNA转染后的成骨分化,随后评估成骨标志物的基因表达水平。然后使用我们的原始阳离子聚合物载体,复合纳米胶束,将mrna施用于大鼠下颌骨制备的骨缺损。采用显微计算机断层扫描(μCT)和组织学分析评估骨再生情况。结果:转染mRNA后,骨钙素(Ocn)、骨桥蛋白(Opn)等成骨标志物显著上调。VEGF mRNA被发现具有与Runx2 mRNA相似的独特成骨功能,两种mRNA联合使用导致标志物进一步上调。在体内给药到骨缺损后,这两种mrna诱导骨再生显著增强,骨矿化增加。使用抗分化簇31蛋白(CD31)、碱性磷酸酶(ALP)或OCN的抗体进行组织学分析显示,mrna诱导缺陷中成骨标志物上调,同时血管形成增加,导致骨快速形成。结论:这些结果证明了利用mRNA药物将包括转录因子在内的各种治疗因子引入靶点的可行性。本研究为组织工程mRNA疗法的发展提供了有价值的信息。
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来源期刊
CiteScore
11.10
自引率
1.20%
发文量
45
审稿时长
11 weeks
期刊介绍: Inflammation and Regeneration is the official journal of the Japanese Society of Inflammation and Regeneration (JSIR). This journal provides an open access forum which covers a wide range of scientific topics in the basic and clinical researches on inflammation and regenerative medicine. It also covers investigations of infectious diseases, including COVID-19 and other emerging infectious diseases, which involve the inflammatory responses. Inflammation and Regeneration publishes papers in the following categories: research article, note, rapid communication, case report, review and clinical drug evaluation.
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