Mineral coated microparticles doped with fluoride and complexed with mRNA prolong transfection in fracture healing

作者：Anna Laura Nelson, Gianluca Fontana, Laura Chubb, Josh Choe, Katherine B. Williams, Daniel P. Regan, Johnny Huard, William L. Murphy, Nicole Ehrhart, Chelsea S. Bahney · 发表于：Frontiers in Bioengineering and Biotechnology · 年份：2024 · DOI：10.3389/fbioe.2023.1295313 · 被引用次数：9 · 研究领域：Bone fractures and treatments、Orthopedic Infections and Treatments、Molecular Biology Techniques and Applications

Introduction: Impaired fracture healing, specifically non-union, has been found to occur up to 14% in tibial shaft fractures. The current standard of care to treat non-union often requires additional surgeries which can result in long recovery times. Injectable-based therapies to accelerate fracture healing have the potential to mitigate the need for additional surgeries. Gene therapies have recently undergone significant advancements due to developments in nanotechnology, which improve mRNA stability while reducing immunogenicity. Methods: In this study, we tested the efficacy of mineral coated microparticles (MCM) and fluoride-doped MCM (FMCM) to effectively deliver firefly luciferase (FLuc) mRNA lipoplexes (LPX) to the fracture site. Here, adult mice underwent a tibia fracture and stabilization method and all treatments were locally injected into the fracture. Level of osteogenesis and amount of bone formation were assessed using gene expression and histomorphometry respectively. Localized and systemic inflammation were measured through gene expression, histopathology scoring and measuring C-reactive protein (CRP) in the serum. Lastly, daily IVIS images were taken to track and measure transfection over time. Results: MCM-LPX-FLuc and FMCM-LPX-FLuc were not found to cause any cytotoxic effects when tested in vitro . When measuring the osteogenic potential of each mineral composition, FMCM-LPX-FLuc trended higher in osteogenic markers through qRT-PCR than the other groups te...