High-glutathione mesenchymal stem cells isolated using the FreSHtracer probe enhance cartilage regeneration in a rabbit chondral defect model.

IF 11.3 1区 医学 Q1 Medicine Biomaterials Research Pub Date : 2023-05-31 DOI:10.1186/s40824-023-00398-3
Gun Hee Cho, Hyun Cheol Bae, Won Young Cho, Eui Man Jeong, Hee Jung Park, Ha Ru Yang, Sun Young Wang, You Jung Kim, Dong Myung Shin, Hyung Min Chung, In Gyu Kim, Hyuk-Soo Han
{"title":"High-glutathione mesenchymal stem cells isolated using the FreSHtracer probe enhance cartilage regeneration in a rabbit chondral defect model.","authors":"Gun Hee Cho, Hyun Cheol Bae, Won Young Cho, Eui Man Jeong, Hee Jung Park, Ha Ru Yang, Sun Young Wang, You Jung Kim, Dong Myung Shin, Hyung Min Chung, In Gyu Kim, Hyuk-Soo Han","doi":"10.1186/s40824-023-00398-3","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Mesenchymal stem cells (MSCs) are a promising cell source for cartilage regeneration. However, the function of MSC can vary according to cell culture conditions, donor age, and heterogeneity of the MSC population, resulting in unregulated MSC quality control. To overcome these limitations, we previously developed a fluorescent real-time thiol tracer (FreSHtracer) that monitors cellular levels of glutathione (GSH), which are known to be closely associated with stem cell function. In this study, we investigated whether using FreSHtracer could selectively separate high-functioning MSCs based on GSH levels and evaluated the chondrogenic potential of MSCs with high GSH levels to repair cartilage defects in vivo.</p><p><strong>Methods: </strong>Flow cytometry was conducted on FreSHtracer-loaded MSCs to select cells according to their GSH levels. To determine the function of FreSHtracer-isolated MSCs, mRNA expression, migration, and CFU assays were conducted. The MSCs underwent chondrogenic differentiation, followed by analysis of chondrogenic-related gene expression. For in vivo assessment, MSCs with different cellular GSH levels or cell culture densities were injected in a rabbit chondral defect model, followed by histological analysis of cartilage-regenerated defect sites.</p><p><strong>Results: </strong>FreSHtracer successfully isolated MSCs according to GSH levels. MSCs with high cellular GSH levels showed enhanced MSC function, including stem cell marker mRNA expression, migration, CFU, and oxidant resistance. Regardless of the stem cell tissue source, FreSHtracer selectively isolated MSCs with high GSH levels and high functionality. The in vitro chondrogenic potential was the highest in pellets generated by MSCs with high GSH levels, with increased ECM formation and chondrogenic marker expression. Furthermore, the MSCs' function was dependent on cell culture conditions, with relatively higher cell culture densities resulting in higher GSH levels. In vivo, improved cartilage repair was achieved by articular injection of MSCs with high levels of cellular GSH and MSCs cultured under high-density conditions, as confirmed by Collagen type 2 IHC, Safranin-O staining and O'Driscoll scores showing that more hyaline cartilage was formed on the defects.</p><p><strong>Conclusion: </strong>FreSHtracer selectively isolates highly functional MSCs that have enhanced in vitro chondrogenesis and in vivo hyaline cartilage regeneration, which can ultimately overcome the current limitations of MSC therapy.</p>","PeriodicalId":9079,"journal":{"name":"Biomaterials Research","volume":null,"pages":null},"PeriodicalIF":11.3000,"publicationDate":"2023-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10233867/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomaterials Research","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1186/s40824-023-00398-3","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0

Abstract

Background: Mesenchymal stem cells (MSCs) are a promising cell source for cartilage regeneration. However, the function of MSC can vary according to cell culture conditions, donor age, and heterogeneity of the MSC population, resulting in unregulated MSC quality control. To overcome these limitations, we previously developed a fluorescent real-time thiol tracer (FreSHtracer) that monitors cellular levels of glutathione (GSH), which are known to be closely associated with stem cell function. In this study, we investigated whether using FreSHtracer could selectively separate high-functioning MSCs based on GSH levels and evaluated the chondrogenic potential of MSCs with high GSH levels to repair cartilage defects in vivo.

Methods: Flow cytometry was conducted on FreSHtracer-loaded MSCs to select cells according to their GSH levels. To determine the function of FreSHtracer-isolated MSCs, mRNA expression, migration, and CFU assays were conducted. The MSCs underwent chondrogenic differentiation, followed by analysis of chondrogenic-related gene expression. For in vivo assessment, MSCs with different cellular GSH levels or cell culture densities were injected in a rabbit chondral defect model, followed by histological analysis of cartilage-regenerated defect sites.

Results: FreSHtracer successfully isolated MSCs according to GSH levels. MSCs with high cellular GSH levels showed enhanced MSC function, including stem cell marker mRNA expression, migration, CFU, and oxidant resistance. Regardless of the stem cell tissue source, FreSHtracer selectively isolated MSCs with high GSH levels and high functionality. The in vitro chondrogenic potential was the highest in pellets generated by MSCs with high GSH levels, with increased ECM formation and chondrogenic marker expression. Furthermore, the MSCs' function was dependent on cell culture conditions, with relatively higher cell culture densities resulting in higher GSH levels. In vivo, improved cartilage repair was achieved by articular injection of MSCs with high levels of cellular GSH and MSCs cultured under high-density conditions, as confirmed by Collagen type 2 IHC, Safranin-O staining and O'Driscoll scores showing that more hyaline cartilage was formed on the defects.

Conclusion: FreSHtracer selectively isolates highly functional MSCs that have enhanced in vitro chondrogenesis and in vivo hyaline cartilage regeneration, which can ultimately overcome the current limitations of MSC therapy.

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
在兔软骨缺损模型中,使用FreSHtracer探针分离的高谷胱甘肽间充质干细胞增强软骨再生。
背景:间充质干细胞(MSCs)是一种很有前途的软骨再生细胞来源。然而,MSC的功能可能因细胞培养条件、供体年龄和MSC群体的异质性而异,导致MSC质量控制不受监管。为了克服这些限制,我们之前开发了一种荧光实时硫醇示踪剂(FreSHtracer),用于监测谷胱甘肽(GSH)的细胞水平,已知谷胱甘肽与干细胞功能密切相关。在本研究中,我们研究了使用FreSHtracer是否可以根据GSH水平选择性分离高功能MSCs,并评估了具有高GSH水平的MSCs在体内修复软骨缺损的软骨形成潜力。方法:用流式细胞仪对负载FreSH示踪剂的MSCs进行流式细胞术,根据其GSH水平选择细胞。为了确定FreSHtracer分离的MSCs的功能,进行了mRNA表达、迁移和CFU测定。MSCs进行软骨分化,然后分析软骨相关基因的表达。为了进行体内评估,将具有不同细胞GSH水平或细胞培养密度的MSCs注射到兔软骨缺损模型中,然后对软骨再生缺损部位进行组织学分析。结果:FreSHtracer根据GSH水平成功分离出MSCs。具有高细胞GSH水平的MSC显示出增强的MSC功能,包括干细胞标记物mRNA表达、迁移、CFU和抗氧化性。无论干细胞组织来源如何,FreSHtracer都选择性地分离出具有高GSH水平和高功能的MSC。在具有高GSH水平的MSCs产生的颗粒中,体外软骨形成潜力最高,ECM形成和软骨形成标记物表达增加。此外,MSC的功能取决于细胞培养条件,相对较高的细胞培养密度导致较高的GSH水平。在体内,通过关节注射具有高水平细胞GSH的MSCs和在高密度条件下培养的MSCs来改善软骨修复,如2型胶原IHC、番红-O染色和奥德里斯科评分所证实的,显示在缺陷上形成了更多的透明软骨。结论:FreSHtracer选择性分离出具有增强体外软骨生成和体内透明软骨再生的高功能MSC,最终可以克服MSC治疗的局限性。
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
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
期刊最新文献
Injectable biomimetic hydrogel constructs for cell-based menopausal hormone therapy with reduced breast cancer potential Targeted H2S-mediated gas therapy with pH-sensitive release property for myocardial ischemia-reperfusion injury by platelet membrane Ultrasound Controllable Release of Proteolysis Targeting Chimeras for Triple Negative Breast Cancer Treatment Multifunctional hydrogels based on γ-polyglutamic acid/polyethyleneimine for hemostasis and wound healing Combining gut microbiota modulation and enzymatic-triggered colonic delivery by prebiotic nanoparticles improves mouse colitis therapy
×
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