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Abstract <p><b>Objective:</b> Exosomes (Exo) have emerged as a promising alternative strategy for treatment of bone defects. The efficacy of exosomes derived from the donor cells is largely influenced by the condition under which the cells are cultured. Recently, exosomes derived from osteogenically induced MC3T3-E1 cells stimulated with SPX (spexin) (OM-Exo-SPX) have shown to promote cell viability, osteoblast differentiation and matrix mineralization <i>in vitro</i>. Therefore, the aim of this study is to explore the effect of OM-Exo-SPX and exosomes derived from osteogenically induced MC3T3-E1 cells (OM-Exo) in bone regeneration. <b>Methods:</b> The mean size and surface protein markers of OM-Exo-SPX and OM-Exo were evaluated using nanoparticle tracking analysis (NTA) and western blot, respectively. Exosome-mediated matrix mineralization was assessed by alizarin red S (ARS) staining. <i>In vivo</i> bone regeneration efficacy of OM-Exo-SPX and OM-Exo was determined using micro-computed tomography (Micro-CT), hematoxylin and eosin (H&amp;E) staining and Masson’s Trichrome staining. <b>Results:</b> SPX markedly increased the release of exosomes from osteogenically induced MC3T3-E1 cells compared to SPX untreated group (<i>p</i> &lt; 0.05). Matrix mineralization effect of OM-Exo-SPX (10 µg/mL) was the highest compared with OM-Exo (10 µg/mL) (<i>p</i> &lt; 0.01) and exosomes untreated group (<i>p</i> &lt; 0.001). Micro-CT, H&amp;E staining and Masson’s Trichrome staining revealed that OM-Exo-SPX (10 µg/µL) significantly promoted bone regeneration compared with OM-Exo and control group. Similarly, OM-Exo displayed more new bone formation at the defect site compared with the control group. <b>Conclusion:</b> Overall, the study discloses OM-Exo-SPX improves <i>in vivo</i> bone regeneration in a calvarial defect model. It also underscores the promise of OM-Exo-SPX as new approach for bone regeneration.</p>

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Exosomes Derived from Spexin-Treated Osteogenic MC3T3-E1 Cells Promote Bone Regeneration in a Calvarial Defect Model

  • 期刊论文

  • 发表日期：

    2026年3月11日

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  • Freshet Assefa,
  • Alemneh Kabeta Daba

摘要

Abstract

Objective: Exosomes (Exo) have emerged as a promising alternative strategy for treatment of bone defects. The efficacy of exosomes derived from the donor cells is largely influenced by the condition under which the cells are cultured. Recently, exosomes derived from osteogenically induced MC3T3-E1 cells stimulated with SPX (spexin) (OM-Exo-SPX) have shown to promote cell viability, osteoblast differentiation and matrix mineralization in vitro. Therefore, the aim of this study is to explore the effect of OM-Exo-SPX and exosomes derived from osteogenically induced MC3T3-E1 cells (OM-Exo) in bone regeneration. Methods: The mean size and surface protein markers of OM-Exo-SPX and OM-Exo were evaluated using nanoparticle tracking analysis (NTA) and western blot, respectively. Exosome-mediated matrix mineralization was assessed by alizarin red S (ARS) staining. In vivo bone regeneration efficacy of OM-Exo-SPX and OM-Exo was determined using micro-computed tomography (Micro-CT), hematoxylin and eosin (H&E) staining and Masson’s Trichrome staining. Results: SPX markedly increased the release of exosomes from osteogenically induced MC3T3-E1 cells compared to SPX untreated group (p < 0.05). Matrix mineralization effect of OM-Exo-SPX (10 µg/mL) was the highest compared with OM-Exo (10 µg/mL) (p < 0.01) and exosomes untreated group (p < 0.001). Micro-CT, H&E staining and Masson’s Trichrome staining revealed that OM-Exo-SPX (10 µg/µL) significantly promoted bone regeneration compared with OM-Exo and control group. Similarly, OM-Exo displayed more new bone formation at the defect site compared with the control group. Conclusion: Overall, the study discloses OM-Exo-SPX improves in vivo bone regeneration in a calvarial defect model. It also underscores the promise of OM-Exo-SPX as new approach for bone regeneration.