Ascorbic Acid-PEI Carbon Dots with Osteogenic Effects as miR-2861 Carriers to Effectively Enhance Bone Regeneration
Ascorbic Acid-PEI Carbon Dots with Osteogenic Effects as miR-2861 Carriers to Effectively Enhance Bone Regeneration
复制标题
具有成骨作用的抗坏血酸-PEI碳点作为miR-2861载体，有效增强骨再生
DOI：
10.1021/acsami.0c15425
复制
发表时间：
2020-11-11
影响因子：
9.5
通讯作者：
Yang, Bai
中科院分区：
文献类型：
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作者：
Bu, Wenhuan;Xu, Xiaowei;Yang, Bai
核酸转移因其持久的局部效应和较低的成本，在骨损伤的治疗中显示出巨大的潜力。纳米材料等具有较高生物相容性的非病毒载体越来越多地应用于骨缺损修复的研究中。表面具有不同活性基团的碳点不仅为携带治疗基因提供了独特的表面，而且一些碳点还被报道促进成骨分化。然而，碳点在体内的骨再生效应很少被研究。MIR-2861具有成骨分化作用。在本研究中，我们通过微波辅助热解的方法制备了能够携带miR-2861的抗坏血酸-PEI碳点(CD)。结果表明，Cd具有良好的荧光稳定性，在体内外具有良好的荧光成像性能。Cd通过网织蛋白介导的内吞途径有效地内化到骨髓基质细胞(BMSCs)中，并分布在线粒体、内质网、溶酶体和细胞核中。碱性磷酸酶染色、茜素红染色和逆转录-实时聚合酶链式反应(RT-QPCR)结果表明，所合成的CD在体外确实具有成骨作用。流式细胞仪检测结果表明，Cd能有效地将miR-2861导入BMSCs，其中携带miR-2861(Cd@miR)的Cd成骨作用最强。血液学、血清生化和组织学分析表明，Cd和Cd@miR在体内无细胞毒性，具有较高的生物相容性。更有趣的是，Cd@miR中的Cd和miR-2861在体外可以协同促进成骨分化，在体内可以显著促进新骨再生。结果表明，携带成骨治疗基因miR-2861的成骨CD具有较强的骨再生能力，具有较大的临床应用潜力。
Nucleic acid transfer has shown significant potential in the treatment of bone damage because of its long lasting local effect and lower cost. Nonviral vectors, such as nanomaterials, with higher biocompatibility are increasedly applied in the study of bone defect repair. Carbon dots with various reactive groups on the surface not only provide a unique surface to carry therapeutic genes, but also some carbon dots have been reported to promote osteogenic differentiation. The bone regeneration effect of carbon dots in vivo, however, is rarely investigated. MiR-2861 has revealed osteogenic differentiation effects. In the current study, we created ascorbic acid-PEI carbon dots (CD), which were able to carry miR-2861, by the microwave-assisted pyrolysis method. Results demonstrated that CD had excellent fluorescence stability leading to good fluorescence imaging in vitro and in vivo. CD was efficiently internalized into bone marrow stromal cells (BMSCs) through the clathrin-mediated endocytosis pathway and distributed in the mitochondria, endoplasmic reticulum, lysosome, and nucleus. Results from alkaline phosphatase staining, alizarin red staining, and reverse transcription real-time PCR (RT-QPCR) showed that our CD indeed had osteogenic effects in vitro. Flow cytometry data indicated that CD could efficiently deliver miR-2861 into BMSCs in vitro, and CD carrying miR-2861 (CD@miR) had the strongest osteogenic effects. Analyses of hematology, serum biochemistry, and histology showed that CD and CD@miR did not have cytotoxicity and had higher biocompatibility in vivo. Most interestingly, CD and miR-2861 in the CD@miR could act synergistically to promote osteogenic differentiation in vitro and new bone regeneration in vivo remarkably. Our results clearly indicate that the osteogenic CD delivering osteogenic therapeutic gene, miR-2861, can obtain much stronger bone regeneration ability, suggesting that our CD has great potential in future clinical application.