Mineral coated microparticles doped with fluoride and complexed with mRNA prolong transfection in fracture healing.
Mineral coated microparticles doped with fluoride and complexed with mRNA prolong transfection in fracture healing.
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DOI：
10.3389/fbioe.2023.1295313
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发表时间：
2023
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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作者：
导读：骨折愈合受损，特别是不愈合，已发现发生高达14%的胫骨干骨折。目前治疗骨不连的护理标准通常需要额外的手术，这可能导致较长的恢复时间。加速骨折愈合的注射疗法有可能减少额外手术的需要。由于纳米技术的发展，基因治疗最近取得了重大进展，纳米技术提高了mRNA的稳定性，同时降低了免疫原性。方法：在本研究中，我们测试了矿物包覆微颗粒（MCM）和氟化物掺杂微颗粒（FMCM）有效递送萤火虫荧光素酶（FLuc） mRNA脂质体（LPX）到骨折部位的效果。在这里，成年小鼠接受胫骨骨折和稳定方法，所有治疗方法都局部注射到骨折中。分别用基因表达法和组织形态学法评估成骨水平和成骨量。通过基因表达、组织病理学评分和血清c反应蛋白（CRP）测定来检测局部和全身炎症。最后，每天拍摄IVIS图像来跟踪和测量转染情况。结果：MCM-LPX-FLuc和FMCM-LPX-FLuc在体外实验中未发现任何细胞毒性作用。在测量每种矿物成分的成骨潜能时，通过qRT-PCR， FMCM-LPX-FLuc在成骨标志物中的含量高于小鼠骨折和稳定模型中测试的其他组。尽管fmcm - lx - fluc显示骨折愈伤组织中il-1β和il-4水平略有升高，但骨折愈伤组织的炎症评分没有任何差异。此外，在任何测试样本中均未观察到急性全身性炎症反应。通过体视学原理分析，小鼠骨折模型中使用的MCM-LPX-FLuc和FMCM-LPX-FLuc浓度对骨骼没有刺激作用。转染效果和传递平台动力学表明，FMCM-LPX-FLuc在体外和体内均延长了荧光素酶信号。讨论：这些数据共同揭示了FMCM-LPX-FLuc可以作为一种有前途的mRNA传递平台用于骨折愈合应用。
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 tested in a murine fracture and stabilization model. Despite FMCM-LPX-FLuc showing slightly elevated il-1β and il-4 levels in the fracture callus, inflammation scoring of the fracture callus did not result in any differences. Additionally, an acute systemic inflammatory response was not observed in any of the samples tested. The concentration of MCM-LPX-FLuc and FMCM-LPX-FLuc that was used in the murine fracture model did not stimulate bone when analyzed through stereological principles. Transfection efficacy and kinetics of delivery platforms revealed that FMCM-LPX-FLuc prolongs the luciferase signal both in vitro and in vivo. Discussion: These data together reveal that FMCM-LPX-FLuc could serve as a promising mRNA delivery platform for fracture healing applications.
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影响因子：
9.7
作者：
Choi, Siyoung;Murphy, William L.
通讯作者：
Murphy, William L.
影响因子：
4.6
作者：
Choi, Siyoung;Yu, Xiaohua;Jongpaiboonkit, Leenaporn;Hollister, Scott J.;Murphy, William L.
通讯作者：
Murphy, William L.
影响因子：
4.6
作者：
Khalil AS;Yu X;Xie AW;Fontana G;Umhoefer JM;Johnson HJ;Hookway TA;McDevitt TC;Murphy WL
通讯作者：
Murphy WL
影响因子：
3.5
作者：
Mukherjee S;Thrasher AJ
通讯作者：
Thrasher AJ