Holkar Ketki, Pethe Prasad, Kale Vaijayanti, Ingavle Ganesh
Symbiosis Centre for Stem Cell Research (SCSCR), Symbiosis School of Biological Sciences (SSBS), Symbiosis International, Deemed University, Lavale, Pune, India.
Clinical Research Facility, NIHR Biomedical Research Centre, Guy's & St Thomas' NHS Foundation Trust and King's College London, Guy's Hospital, London, UK.
Regen Med. 2025 Sep 8:1-12. doi: 10.1080/17460751.2025.2557770.
This study aimed to enhance the osteoinductive potential of mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) by integrating them into a nano-hydroxyapatite (nHAp)-enriched hydrogel scaffold for bone regeneration applications.
MATERIALS & METHODS: EVs were isolated from naïve and osteogenically primed MSCs and characterized for morphology, cargo content, and cytocompatibility. Their uptake and osteoinductive activity were assessed using MC3T3 cells within a 3D interpenetrating network (IPN) hydrogel. The most effective EV formulation was incorporated into an nHAp - IPN hydrogel scaffold and evaluated both in vitro and in a murine subcutaneous implantation model.
Primed MSC-EVs showed elevated calcium, ALP activity, and osteogenic/angiogenic mRNAs (, ) compared to naïve EVs, with comparable size and morphology. Both EV types were internalized efficiently without cytotoxicity. In combination with nHAp, primed EVs enhanced ALP activity, calcium deposition, and mineralization. Histological analysis confirmed scaffold biocompatibility and mineralized tissue formation.
Osteogenically primed MSC-EVs significantly improved the osteoinductive performance of nHAp-based hydrogels, supporting their potential as a cell-free therapeutic strategy for bone tissue engineering.
本研究旨在通过将间充质干细胞衍生的细胞外囊泡（MSC-EVs）整合到富含纳米羟基磷灰石（nHAp）的水凝胶支架中，以增强其骨诱导潜力，用于骨再生应用。
从未经处理和经成骨诱导的间充质干细胞中分离出细胞外囊泡，并对其形态、所载成分和细胞相容性进行表征。在三维互穿网络（IPN）水凝胶中使用MC3T3细胞评估它们的摄取和骨诱导活性。将最有效的细胞外囊泡制剂掺入nHAp-IPN水凝胶支架中，并在体外和小鼠皮下植入模型中进行评估。
与未经处理的细胞外囊泡相比，经诱导的MSC-EVs显示出升高的钙、碱性磷酸酶活性和成骨/血管生成mRNA（ ， ），且大小和形态相当。两种类型的细胞外囊泡均能有效内化且无细胞毒性。与nHAp结合时，经诱导的细胞外囊泡增强了碱性磷酸酶活性、钙沉积和矿化。组织学分析证实了支架的生物相容性和矿化组织形成。
经成骨诱导的MSC-EVs显著改善了基于nHAp的水凝胶的骨诱导性能，支持其作为骨组织工程的无细胞治疗策略的潜力。
