Engineering a cell-free bone regeneration platform using osteogenically primed MSC-EVs and nHAp-enriched IPN hydrogels

Sep 8, 2025Regenerative medicine

Building a cell-free bone healing system using bone-promoting cell particles and mineral-enriched supportive gels

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Regenerative Health on OpenScience ↗PubMed ↗DOI ↗

Abstract

Osteogenically primed mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) enhanced osteoinductive activity in a nano-hydroxyapatite hydrogel scaffold.

Primed MSC-EVs exhibited elevated calcium levels and alkaline phosphatase (ALP) activity compared to naïve MSC-EVs.
Both types of EVs were effectively taken up by cells without causing cytotoxicity.
The combination of primed MSC-EVs and the nano-hydroxyapatite scaffold improved mineralization and ALP activity.
Histological analysis indicated that the scaffold was biocompatible and supported mineralized tissue formation.

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AIMS: 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 assessedusing 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. in vitro

RESULTS: 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, andmineralization. Histological analysis confirmed scaffold biocompatibility and mineralized tissue formation. Runx2Vegf-a in vivo

CONCLUSIONS: 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.

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Engineering a cell-free bone regeneration platform using osteogenically primed MSC-EVs and nHAp-enriched IPN hydrogels

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