AimTo identify an optimized strategy for the large‐scale production of nanovesicles (NVs) that preserve the biological properties of exosomes (EXOs) for use in periodontal regeneration.Materials and MethodsNVs from dental follicle stem cells (DFSCs) were prepared through extrusion, and EXOs from DFSCs were isolated. The yield of both extruded NVs (eNVs) and EXOs were quantified through protein concentration and particle number analyses. Their pro‐migration, pro‐proliferation and pro‐osteogenesis capacities were compared subsequently in vitro. Additionally, proteomics analysis was conducted. To further evaluate the periodontal regeneration potential of eNVs and EXOs, they were incorporated into collagen sponges and transplanted into periodontal defects in rats. In vivo imaging and H&E staining were utilized to verify their biodistribution and safety. Micro‐Computed Tomography analysis and histological staining were performed to examine the regeneration of periodontal tissues.ResultsThe yield of eNVs was nearly 40 times higher than that of EXOs. Interestingly, in vitro experiments indicated that the pro‐migration and pro‐proliferation abilities of eNVs were superior, and the pro‐osteogenesis potential was comparable to EXOs. More importantly, eNVs exhibited periodontal regenerative potential similar to that of EXOs.ConclusionsExtrusion has proven to be an efficient method for generating numerous eNVs with the potential to replace EXOs in periodontal regeneration.

中文翻译：

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从牙囊干细胞中挤出的高产量纳米囊泡作为外泌体的替代品促进牙周组织的再生


目的确定一种用于大规模生产纳米囊泡 （NV） 的优化策略，该囊泡保留了外泌体 （EXO） 的生物学特性，用于牙周再生。材料和方法通过挤出制备牙囊干细胞 （DFSC） 的 NVs，并分离 DFSCs 的 EXO。通过蛋白质浓度和颗粒数分析量化挤出 NVs （eNVs） 和 EXOs 的产量。随后在体外比较了它们的促迁移、促增殖和促成骨能力。此外，还进行了蛋白质组学分析。为了进一步评估 eNVs 和 EXOs 的牙周再生潜力，将它们掺入胶原蛋白海绵并移植到大鼠的牙周缺损中。利用体内成像和 H&E 染色验证其生物分布和安全性。进行显微计算机断层扫描分析和组织学染色以检查牙周组织的再生。结果eNVs 的产量是 EXO 的近 40 倍。有趣的是，体外实验表明，eNVs 的促迁移和促增殖能力优越，促成骨潜力与 EXOs 相当。更重要的是，eNVs 表现出与 EXOs 相似的牙周再生潜力.结论挤压已被证明是产生大量 eNVs 的有效方法，有可能在牙周再生中取代 EXOs。