Extracellular Vesicles (EVs) Mirror Their Source: Comparative Analysis of Bone Marrow-Derived and Adipose-Derived Stem Cells and EVs in Bone Regeneration.
{"title":"Extracellular Vesicles (EVs) Mirror Their Source: Comparative Analysis of Bone Marrow-Derived and Adipose-Derived Stem Cells and EVs in Bone Regeneration.","authors":"Chun-Yee Ho, Shih-Hsuan Mao, Chia-Hsuan Tsai, Kai-Ti Chuang, Bo-Ru Lai, Sheng-Wei Wang, Chih-Hao Chen, Chien-Tzung Chen","doi":"10.1007/s12195-026-00909-x","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Critical-sized craniofacial defects pose a significant clinical challenge, prompting the investigation of novel regenerative strategies. While mesenchymal stem cells (MSCs) and extracellular vesicles (EVs) hold promise, the optimal cell source and EV efficacy for craniofacial bone regeneration remain unclear. This study compares adipose-derived stem cells (ASCs), bone marrow-derived stem cells (BMSCs), and their derived EVs to address this gap in a critical-sized calvarial defect model.</p><p><strong>Methods: </strong>EVs from BMSCs and ASCs were isolated via ultracentrifugation and size exclusion chromatography. Nanoparticle tracking analysis and bicinchoninic acid assay quantified yield and protein, respectively. Transmission electron microscopy and Western blotting verified EV morphology and markers. In vitro, osteogenic potential of BMSCs and ASCs treated with their respective EVs was assessed using alkaline phosphatase activity assay, viability assays, and mineralization staining. In vivo, bone regeneration was compared in a rat critical-sized calvarial defect model treated with BMSCs, BMSC-derived EVs, and a combination of BMSC and BMSC-derived EVs.</p><p><strong>Results: </strong>EVs isolated by ultracentrifugation yielded superior numbers of particles compared to size exclusion chromatography. In vitro, BMSC-derived EVs enhanced osteogenic differentiation of BMSCs, whereas ASC-derived EVs inhibited proliferation and osteogenesis of BMSCs. Although BMSC-derived EVs induced osteogenic phenotype in ASCs, osteoinductive efficiency was low, along with reduced cell proliferation. In vivo, both BMSC-derived EVs and BMSCs individually promoted bone regeneration compared to vehicle controls. Notably, the combination of BMSCs and BMSC-derived EVs demonstrated a significantly superior healing within the bone defect.</p><p><strong>Conclusions: </strong>Ultracentrifugation is the preferred method for isolating EVs for clinical translation. BMSC-derived EVs are the optimal source for craniofacial bone regeneration compared to ASC-derived EVs, as they exhibited superior osteogenic potential and promoted bone regeneration. Moreover, BMSC-derived EVs combined with BMSCs demonstrated a synergistic effect that further improved bone regeneration. Notably, mismatching origins of MSC and MSC-derived EVs could dysregulate the cellular function of MSCs, potentially compromising the regenerative outcomes. Our study highlights that the proper matching of cell and EVs sources is imperative for optimizing therapeutic efficacy in craniofacial bone regeneration.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-026-00909-x.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 3","pages":"297-310"},"PeriodicalIF":3.5000,"publicationDate":"2026-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13365279/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cellular and molecular bioengineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s12195-026-00909-x","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2026/6/1 0:00:00","PubModel":"eCollection","JCR":"Q3","JCRName":"BIOPHYSICS","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Critical-sized craniofacial defects pose a significant clinical challenge, prompting the investigation of novel regenerative strategies. While mesenchymal stem cells (MSCs) and extracellular vesicles (EVs) hold promise, the optimal cell source and EV efficacy for craniofacial bone regeneration remain unclear. This study compares adipose-derived stem cells (ASCs), bone marrow-derived stem cells (BMSCs), and their derived EVs to address this gap in a critical-sized calvarial defect model.
Methods: EVs from BMSCs and ASCs were isolated via ultracentrifugation and size exclusion chromatography. Nanoparticle tracking analysis and bicinchoninic acid assay quantified yield and protein, respectively. Transmission electron microscopy and Western blotting verified EV morphology and markers. In vitro, osteogenic potential of BMSCs and ASCs treated with their respective EVs was assessed using alkaline phosphatase activity assay, viability assays, and mineralization staining. In vivo, bone regeneration was compared in a rat critical-sized calvarial defect model treated with BMSCs, BMSC-derived EVs, and a combination of BMSC and BMSC-derived EVs.
Results: EVs isolated by ultracentrifugation yielded superior numbers of particles compared to size exclusion chromatography. In vitro, BMSC-derived EVs enhanced osteogenic differentiation of BMSCs, whereas ASC-derived EVs inhibited proliferation and osteogenesis of BMSCs. Although BMSC-derived EVs induced osteogenic phenotype in ASCs, osteoinductive efficiency was low, along with reduced cell proliferation. In vivo, both BMSC-derived EVs and BMSCs individually promoted bone regeneration compared to vehicle controls. Notably, the combination of BMSCs and BMSC-derived EVs demonstrated a significantly superior healing within the bone defect.
Conclusions: Ultracentrifugation is the preferred method for isolating EVs for clinical translation. BMSC-derived EVs are the optimal source for craniofacial bone regeneration compared to ASC-derived EVs, as they exhibited superior osteogenic potential and promoted bone regeneration. Moreover, BMSC-derived EVs combined with BMSCs demonstrated a synergistic effect that further improved bone regeneration. Notably, mismatching origins of MSC and MSC-derived EVs could dysregulate the cellular function of MSCs, potentially compromising the regenerative outcomes. Our study highlights that the proper matching of cell and EVs sources is imperative for optimizing therapeutic efficacy in craniofacial bone regeneration.
Supplementary information: The online version contains supplementary material available at 10.1007/s12195-026-00909-x.
期刊介绍：
The field of cellular and molecular bioengineering seeks to understand, so that we may ultimately control, the mechanical, chemical, and electrical processes of the cell. A key challenge in improving human health is to understand how cellular behavior arises from molecular-level interactions. CMBE, an official journal of the Biomedical Engineering Society, publishes original research and review papers in the following seven general areas:
Molecular: DNA-protein/RNA-protein interactions, protein folding and function, protein-protein and receptor-ligand interactions, lipids, polysaccharides, molecular motors, and the biophysics of macromolecules that function as therapeutics or engineered matrices, for example.
Cellular: Studies of how cells sense physicochemical events surrounding and within cells, and how cells transduce these events into biological responses. Specific cell processes of interest include cell growth, differentiation, migration, signal transduction, protein secretion and transport, gene expression and regulation, and cell-matrix interactions.
Mechanobiology: The mechanical properties of cells and biomolecules, cellular/molecular force generation and adhesion, the response of cells to their mechanical microenvironment, and mechanotransduction in response to various physical forces such as fluid shear stress.
Nanomedicine: The engineering of nanoparticles for advanced drug delivery and molecular imaging applications, with particular focus on the interaction of such particles with living cells. Also, the application of nanostructured materials to control the behavior of cells and biomolecules.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
