In Vivo Bone Tissue Induction by Freeze-dried Collagen-nanohydroxyapatite Matrix Loaded with BMP2/NS1 mRNAs Lipopolyplexes
Résumé
Messenger RNA (mRNA) activated matrices (RAMs) are interesting for bone regeneration since they have the potentiality for in-situ and sustained production of osteogenic proteins. The main obstacles of mRNA therapy are the activation of RNA sensors and lacking of proper in vivo mRNA delivery vector. In a previous study, we developed a dual mRNAs system for in vitro osteogenesis by co-delivering BMP2 mRNA and NS1 mRNA which effectively inhibited RNA sensors and enhanced BMP-2 expression. Here, we evaluated a lipopolyplex (LPR) platform for in vivo mRNA delivery and adapted the BMP2/NS1 mRNA LPRs, in form of RAM, for in vivo bone induction. Through intradermal injection, the firefly luciferase mRNA LPRs generated strong bioluminescence signal. Then, the BMP2/NS1 mRNAs LPRs were incorporated into a collagen-nanohydroxyapatite scaffold and freeze-dried to prepare ready-to-use RAMs. Scanning electron microscopy imaging demonstrated that the loaded LPRs maintained their spherical morphology in the RAM, thanks to the core-shell structure of the LPRs. The mRNAs in vitro release from RAMs lasted for 16 days and resulted in a 2-fold prolonged protein expression period compared to direct one time single cells transfection. 8 weeks after mice back subcutaneous implantation, the μCT and histology analyses showed that the BMP2/NS1 mRNAs LPRs containing RAMs (RAM-BMP2/NS1) induced significantly higher more new bone tissue than those without NS1 mRNA. Overall, our results demonstrate that the BMP2/NS1 dual mRNAs system is suitable for osteogenic engagement, and the freeze-dried RAM-BMP2/NS1 could be promising off-the-shelf products for clinical orthopedic practice.
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