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
{"title":"Enhancement of bone regeneration by coadministration of angiogenic and osteogenic factors using messenger RNA.","authors":"Maorui Zhang,&nbsp;Yuta Fukushima,&nbsp;Kosuke Nozaki,&nbsp;Hideyuki Nakanishi,&nbsp;Jia Deng,&nbsp;Noriyuki Wakabayashi,&nbsp;Keiji Itaka","doi":"10.1186/s41232-023-00285-3","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":13588,"journal":{"name":"Inflammation and Regeneration","volume":null,"pages":null},"PeriodicalIF":5.0000,"publicationDate":"2023-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10280834/pdf/","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Inflammation and Regeneration","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s41232-023-00285-3","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
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.

Abstract Image

Abstract Image

Abstract Image

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
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.
期刊最新文献
CX3CL1-CX3CR1 axis protects retinal ganglion cells by inhibiting microglia activation in a distal optic nerve trauma model Emilin2 marks the target region for mesenchymal cell accumulation in bone regeneration Role of cellular senescence in inflammation and regeneration Th22 is the effector cell of thymosin β15-induced hair regeneration in mice The gut-liver axis in hepatobiliary diseases
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1