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Background <p>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> Methods <p>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> Results <p>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> Conclusions <p>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>

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Extracellular Vesicles (EVs) Mirror Their Source: Comparative Analysis of Bone Marrow-Derived and Adipose-Derived Stem Cells and EVs in Bone Regeneration

  • 期刊论文

  • 发表日期：

    2026年6月16日

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  • Chun-Yee Ho,
  • Shih-Hsuan Mao,
  • Chia-Hsuan Tsai,
  • Kai-Ti Chuang,
  • Bo-Ru Lai,
  • Sheng-Wei Wang,
  • Chih-Hao Chen,
  • Chien-Tzung Chen

摘要

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.