Zhang Qian, Qu Xingyuan, Liang Chen, Li Hongyan, Du Siyu, Wang Chang, Xie Yuandong, Zheng Yi, Wang Lei
Department of Periodontics, Hospital of Stomatology, Jilin University, Changchun, China.
Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling, Jilin University, Changchun, China.
Front Bioeng Biotechnol. 2023 Jun 19;11:1204571. doi: 10.3389/fbioe.2023.1204571. eCollection 2023.
This study aimed to investigate the regulatory effect of N-isopropylacrylamide-modified polyethyleneimine (PEN)-delivered oligodeoxynucleotide (ODN) MT01 on bone regeneration and . A polyethylenimine (PEI) derivative, PEN, was constructed through Michael addition and employed as a carrier for ODN MT01 transfection. PEN/MT01 nanocomposites were characterized using agarose gel retardation assay, size distribution, zeta potential and transmission electron microscopy. The Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of PEN on cell viability. Alkaline phosphatase (ALP) staining was used to detect the osteogenic differentiation ability of PEN/MT01 nanocomposite. Real-time quantitative PCR (q RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the regulatory effects of PEN/MT01 nanocomposite on osteogenic differentiation gene expression. Rat model was observed using the skull defect method and verified using micro-computed tomography (CT), serum biochemical indices, hematoxylin and eosin (H&E) staining and Immunohistochemistry (IHC). PEN had good biological properties and could deliver MT01 well to achieve efficient transmission of MT01. PEN/MT01 nanocomposites were effectively transfected into MC3T3-E1 cells at a ratio of 6.0. CCK-8 assay displayed that PEN had no cytotoxicity to MC3T3-E1 cells. Additionally, PEN/MT01 nanocomposites could promote the expression of osteogenic genes. results revealed that PEN/MT01 nanocomposites could promote bone regeneration more effectively than the other groups. PEN has good biocompatibility and low toxicity, which is a good carrier for ODN MT01. PEN-delivered MT01 can be potentially employed as a useful approach to achieving bone regeneration.
本研究旨在探讨N-异丙基丙烯酰胺修饰的聚乙烯亚胺（PEN）递送的寡脱氧核苷酸（ODN）MT01对骨再生的调节作用。通过迈克尔加成反应构建了聚乙烯亚胺（PEI）衍生物PEN，并将其用作ODN MT01转染的载体。使用琼脂糖凝胶阻滞试验、尺寸分布、zeta电位和透射电子显微镜对PEN/MT01纳米复合材料进行表征。采用细胞计数试剂盒-8（CCK-8）试验检测PEN对细胞活力的影响。碱性磷酸酶（ALP）染色用于检测PEN/MT01纳米复合材料的成骨分化能力。实时定量聚合酶链反应（q RT-PCR）和酶联免疫吸附测定（ELISA）用于检测PEN/MT01纳米复合材料对成骨分化基因表达的调节作用。采用颅骨缺损法观察大鼠模型，并通过微型计算机断层扫描（CT）、血清生化指标、苏木精-伊红（H&E）染色和免疫组织化学（IHC）进行验证。PEN具有良好的生物学特性，能够很好地递送MT01，实现MT01的高效传递。PEN/MT01纳米复合材料以6.0的比例有效地转染到MC3T3-E1细胞中。CCK-8试验显示PEN对MC3T3-E1细胞无细胞毒性。此外，PEN/MT01纳米复合材料可以促进成骨基因的表达。结果表明，PEN/MT01纳米复合材料比其他组更有效地促进骨再生。PEN具有良好的生物相容性和低毒性，是ODN MT01的良好载体。PEN递送的MT01有可能作为实现骨再生的一种有用方法。
