Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Institute of Stomatological Research, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, People's Republic of China.
Laboratory of Biomaterials, Key Laboratory on Assisted Circulation, Ministry of Health, Cardiovascular Division, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, People's Republic of China.
Int J Nanomedicine. 2019 Jun 6;14:4229-4245. doi: 10.2147/IJN.S203540. eCollection 2019.
Gene therapies via small interfering (si)RNA () and bone morphogenetic protein () plasmid DNA () may be promising strategies for bone repair/regeneration, but their ideal delivery vectors, efficacy difference, and underlying mechanisms have not been explored, so these issues were probed here. This study used lipopolysaccharide-amine nanopolymersomes (LNPs), an efficient cytosolic delivery vector developed by the research team, to mediate and to transfect MC3T3-E1 cells, respectively. The cytotoxicity, cell uptake, and gene knockdown efficiency of -loaded LNPs (LNPs/) were studied, then the osteogenic-differentiation efficacy of MC3T3-E1 cells treated by LNPs/ and LNPs/, respectively, were compared by measuring the expression of osteogenesis-related genes and proteins, alkaline phosphatase (ALP) activity, and mineralization of the extracellular matrix at all osteogenic stages. Finally, the possible signaling pathways of the two treatments were explored. LNPs delivered into cells efficiently to silence 50% of expression without obvious cytotoxicity. LNPs/ and LNPs/ enhanced the osteogenic differentiation of MC3T3 E1 cells, but LNPs/ was better than LNPs/. BMP/Mothers against decapentaplegic homolog (Smad) and glycogen synthase kinase (GSK)-3β/β-catenin signaling pathways appeared to be involved in osteogenic differentiation induced by LNPs/, but GSK-3β/β-catenin was not stimulated upon LNPs/ treatment. LNPs are safe and efficient delivery vectors for DNA and RNA, which may find wide applications in gene therapy. treatment may be a more efficient strategy to enhance osteogenic differentiation than treatment. LNPs loaded with and/or may provide new opportunities for the repair and regeneration of bone.
基于小干扰 RNA()和骨形态发生蛋白()质粒 DNA()的基因治疗可能是骨修复/再生的有前途的策略，但它们的理想递药载体、疗效差异和潜在机制尚未得到探索，因此本研究对此进行了探讨。本研究使用脂多糖-胺纳米聚合物囊泡（LNPs）作为有效的胞质递药载体，该载体由研究团队开发，分别转染 MC3T3-E1 细胞。研究了负载的 LNPs（LNPs/）的细胞毒性、细胞摄取和基因敲低效率，然后通过测量成骨相关基因和蛋白的表达、碱性磷酸酶（ALP）活性以及整个成骨阶段细胞外基质的矿化，比较了 LNPs/和 LNPs/处理的 MC3T3-E1 细胞的成骨分化效果。最后，探索了两种处理方法的可能信号通路。LNPs 有效地将转染到细胞中，使表达沉默了 50%，而没有明显的细胞毒性。LNPs/和 LNPs/增强了 MC3T3 E1 细胞的成骨分化，但 LNPs/优于 LNPs/。BMP/Mothers against decapentaplegic homolog (Smad)和糖原合成酶激酶（GSK）-3β/β-catenin 信号通路似乎参与了 LNPs/诱导的成骨分化，但 GSK-3β/β-catenin 未受到 LNPs/处理的刺激。LNPs 是 DNA 和 RNA 的安全有效的递药载体，可能在基因治疗中有广泛的应用。治疗可能是增强成骨分化比治疗更有效的策略。负载和/或的 LNPs 可能为骨的修复和再生提供新的机会。
